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Microbial treatment for nanocellulose extraction from marine

algae and its applications as sustainable functional material

Muhammad Zaki, H.P.S. Abdul Khalil, F.A. Sabaruddin, R.D.

Bairwan, Adeleke A. Oyekanmi, Tata Alfatah, Mohammed
Danish, E.M. Mistar, C.K. Abdullah

PII: S2589-014X(21)00189-4
DOI: https://doi.org/10.1016/j.biteb.2021.100811
Reference: BITEB 100811

To appear in: Bioresource Technology Reports

Received date: 7 June 2021

Revised date: 23 August 2021
Accepted date: 25 August 2021

Please cite this article as: M. Zaki, H.P.S. Abdul Khalil, F.A. Sabaruddin, et al., Microbial
treatment for nanocellulose extraction from marine algae and its applications as
sustainable functional material, Bioresource Technology Reports (2018), https://doi.org/
10.1016/j.biteb.2021.100811

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© 2018 © 2021 Published by Elsevier Ltd.

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Microbial Treatment for Nanocellulose Extraction from Marine Algae and its Applications as
Sustainable Functional Material

Muhammad Zaki a*, Abdul Khalil H.P.S. b*, F. A. Sabaruddin b, R. D. Bairwan b, Adeleke A.
Oyekanmi b, Tata Alfatah b, Mohammed Danish b, E. M. Mistar b, and C. K. Abdullah b

a
Department of Chemical Engineering, Universitas Syiah Kuala, Banda Aceh 23111, Indonesia
b
School of Industrial Technology, Universiti Sains Malaysia, Penang 11800, Malaysia
*Corresponding email: [email protected] (Abdul Khalil H.P.S); [email protected]
(Muhammad Zaki)

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Abstract
With increasing environmental awareness among the masses, the demand for eco-friendly products

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in the market has increased significantly. The market demand and ecological concerns led
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researchers to develop an eco-friendly material based on biopolymers. Marine algae-derived
nanocellulose has been known to be utilized as a functional material for a wide range of applications,
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such as packaging, environmental, and biomedical applications. Many studies have revealed that the
extraction of nanocellulose from algae biomass can be achieved using microbial, chemical,
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mechanical, or hybrid methods. Microbial treatment via enzymatic hydrolysis uses an enzyme,
namely cellulase. Cellulase can be produced by micro-organisms like bacteria, yeast, and fungi to
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increase production by cutting down on operating costs as they are cheaply available. Recent work
regarding various aspects of microbial nanocellulose production has been discussed. This review also
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highlights the utilization of macroalgae as sustainable, functional materials in a wide range of

applications.
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Keywords: Enzyme hydrolysis, micro-organism, seaweed, microbial nanocellulose, cellulase,

wastewater treatment

1. Introduction
In the past few decades, the development of nanotechnology has witnessed remarkable success,
particularly in the production of nanocellulose. Nanocellulose derived from lignocellulosic materials
is a potential substitute for petroleum-derived feedstock to obtain many commercial products that
comply with market demand and satisfy environmental concerns (Dufresne, 2013; Jaiswal et al.,
2021; Squinca et al., 2020). The prospects of nanocellulose produced from a wide range of natural
resources, including plant biomass, bacteria, and algae, have been widely explored in
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nanotechnology due to their potentials (Liu et al., 2017a). The annual production of cellulose and its
derivatives has been estimated at 180 billion tons, while its market demand has been continuously
increasing in recent years (Baghel et al., 2021; Hafid et al., 2021). Nanocellulose has been reported
to be utilized as a versatile material. It has a wide range of applications in a varied range of
industries, such as the electronic industry (as electronic displays), construction device industry (as
sensors to monitor stress levels for bridges) (Dias et al., 2020; Hsu and Zhong, 2019; Xing et al.,
2019), food packaging materials industry, for example as replacement of synthetic packaging barrier
film (Ahankari et al., 2021), additive for antioxidant and antimicrobial free-standing films (Bastante
et al., 2021; Reshmy et al., 2021), edible and biodegradable packaging films (Kumar et al., 2021;
Zhang et al., 2021), and 3D printed food packahing aerogel (Zhou et al., 2021), paper and coating

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materials industry as additive and paper coating (Abdul Khalil et al., 2017; Li et al., 2021;Sharma et

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al., 2020;Zulfiana et al., 2020), additive in grease proof paper (Hubbe and Pruszynski, 2020),
enzymatic nanocellulose for pulp and paper (Michelin et al., 2020), water treatment industry
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(adsorbent material) (Choudhury et al., 2020; Mautner, 2020; Mehanny et al., 2021; R et al., 2021;
Shang et al., 2021; Tan et al., 2020), medical and tissue engineering industry as supporting materials
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in water absorbent pads (Ebadi et al., 2021), wound dressing (Claro et al., 2020; Resch et al., 2021),
drug release devices manufacturing (Venkatesan et al., 2016), and scaffold for tissue engineering
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(Abdul Khalil et al., 2020).

In general, nanocellulose can be categorized into three main types, known as nanocrystalline
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cellulose (NCC), nanofibrillated cellulose (NFC), and bacterial cellulose (BC). These nanocelluloses
can be produced via two approaches: top-down and bottom-up. NCC and NFC are examples of
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nanocellulose using the top-down approach, while bacterial cellulose is made using the bottom-up
approach (Klemm et al., 2011; Phanthong et al., 2018). The top-down approach indicates the
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disintegration of lignocellulosic biomass into the nanofiber via chemical or mechanical processes
(Lee et al., 2012). The cellulose extraction is initiated by removing the amorphous part of the fibers
through the pulping and bleaching process. The purified cellulose is then subjected to the mechanical
process, such as using grinders, high-pressure homogenizers, etc. This high shear fibrillation process
enables converting micrometer-sized cellulose into nanocellulose, which is referred to as
nanofibrillated cellulose (NFC) (Lee et al., 2012).
Meanwhile, the cellulose fiber undergoes a hydrolysis process known as nanocrystalline
cellulose (NCC). This process involves the application of strong acids like hydrochloric acid (HCl)
or sulfuric acid (H2SO4) to hydrolyze the amorphous segments of cellulose fibers to achieve a perfect
nano-whisker-like nanocellulose fiber with high crystallinity properties (Sabaruddin et al., 2020).
However, the extensive use of chemicals during the process needs to be removed to achieve pure
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nanocellulose free from any by-products that can deteriorate its properties (Sabaruddin et al., 2021;
Wahlström et al., 2020). In addition, these procedures are energy-intensive, economically non-viable,
and require plenty of water in the neutralization step, which increases the environmental pollution
load. Besides, the extensive use of chemicals during the purification process can cause a potential
secondary pollution problem. For example, during the pre-treatment process, such as the pulping
process that involves the applications of large quantities of chemicals of which sodium hydroxide is
the most effective, were seen to cause potential environmental hazards. In addition to that, the pre-
treatment process of nanocellulose production also required a bleaching process, and it is considered
an important pre-treatment step. This process requires a number of chemicals, including chlorine
dioxide (ClO2), hydrogen peroxide (H2O2), sodium hypochlorite (NaClO), ozone (O3), and peracetic

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acid (CH3CO3H). These chemicals were used during the delignification process to remove the

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residual lignin fractions and were usually repeated several times to achieve fully purified cellulose
fibers. Therefore, this method is considered highly toxic and enables the production of hazardous
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pollutants. To date, many alternatives bleaching procedures have been introduced, for instance,
elemental chlorine-free (ECF) and total chlorine-free (TCF) to replace the conventional
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delignification process (Dhali et al., 2021). Therefore, different pre-treatment protocols that can
promote more environmentally friendly effects need to be introduced. This is because applying these
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extensive chemicals required a thorough neutralization, and this process needs huge water utilization
for the washing process. This process leads to an extensive amount of waste and leads to economic
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and environmental constraints that make nanocellulose industries underdeveloped (Ribeiro et al.,
2019; Shen et al., 2020). The production of bacterial cellulose through bottom-up approaches, on the
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other hand, is different compared to that of NCC and NFC. BC is excreted by bacteria such as
Acetobacter sp. to produce nanocellulose. The process involves fermentation of low molecular sugar
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using cellulose-producing bacteria to produce pure cellulose without the presence of pectin and
lignin (Lee et al., 2012).
Recently, microbial-treated nanocellulose via the processes of enzymatic hydrolysis,
fermentation, fungi degrading, etc., has shown promising environmental-friendly for sustainable
growth. The enzymatic hydrolysis possesses a high substrate selectivity that targets specific
lignocellulosic linkages, which promote toxic free residue formation and performs under mild
conditions that result in a less energy-intensive process (Fritz et al., 2015). Enzymatic hydrolysis
uses an enzyme, namely cellulase, that catalyzes the breakdown of the carbohydrate cell wall and
assists in the isolation of nanocellulose (Irawan et al., 2021; Singh et al., 2021). A growing number
of publications related to nanocellulose production via bioconversion have been reported in the
recent past, mainly focusing on improving the production process to develop nanocellulose
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composites. A recent review article by Ribeiro et al. (2019) has reported a remarkable increment in
the number of publications primarily related to the enzymatic production of nanocellulose. Most of
the research is mainly devoted to identifying enzymes for efficient cellulose degradation,
mechanisms, and efficient processes for de-fibrillating cellulosic materials (Jaiswal et al., 2021).
These works can contribute to the potential of novel and efficient nanocellulose production methods
and innovative applications for the next few years.
To date, increasing attention has been given to the cellulose derived from marine biomass,
mainly algae, due to its potential benefits compared to cellulose from inland biomass (Chen et al.,
2016; Wahlström et al., 2020). Heretofore, marine algae have been utilized for food supplements,
pharmaceuticals, and dyes. Their unique properties enable them to be widely studied in a multitude

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of arena that promise to be beneficial for industrial applications (Beetul et al., 2016). Today, marine

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biomass is considered a promising future source for the isolation of nanocellulose because of the
absence of lignin in marine algae. The lignin-free biomass imparts more benefits in nanocellulose
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extraction compared to lignocellulosic biomass. Furthermore, the abundance of pure cellulose and
the lack of lignin in marine algae are potentially better suited for high-quality nanocellulose
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extraction. Besides, this nanocellulose has a broader range of applications than the nanocellulose
extracted from lignocellulosic biomass. Moreover, the abundance and availability of algae biomass,
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either in micro or macro form, has a promising prospect for the extraction of nanocellulose (Chen et
al., 2016; Ilyas and Atikah, 2021; Korzen et al., 2015). The utilization of marine algae biomass has
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promising prospects due to its advantages of having a low content of natural physicochemical
barriers, higher yields of stored carbohydrates, the capacity to grow under normal growth conditions,
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and a rapid growth rate within a short period compared to other inland biomass (Paniz et al., 2020).
Despite the simple extraction process, the algae-derived nanocellulose has been reported to
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demonstrate a higher degree of crystallinity, specific surface area, and excellent rheological and
mechanical properties compared to forest waste and agriculture residues derived nanocellulose (Yu
et al., 2021).
Regardless of the remarkable potential of marine algae biomass, a comprehensive study that
focuses on the microbial extraction of nanocellulose from algae biomass and its application as a
functional material for a wide range of applications has not been reported so far. Therefore, the scope
of this study encompasses sustainability factors for the abundance of algae biomass as a precursor for
the extraction of nanocellulose. Detailed methods of extraction of nanocellulose and properties as a
functional material for packaging environmental and biomedical applications are included in the
study. Finally, this study concluded with future challenges and prospects of sustainable marine algae-
derived nanocellulose and its environmentally friendly applications.
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2. Marine algae as a source of nanocellulose

Marine algae are considered important organisms for their habitats and have been recognized even
more in green technology. Their utilization in human society has been dated back to 500 B.C. in
China and later spread all across the globe, which continued to be applied until today (Beetul et al.,
2016). The ocean covers more than 71% of the world’s surface, serving as the habitat for most marine
organisms, including marine algae species. Ecologically, marine algae are the basis of most aquatic
food chains and have been known as the most abundant resource in the ocean. Marine algae contain a

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rich source of functional metabolites, including polysaccharides, proteins, peptides, lipids, amino

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acids, polyphenols, and mineral salts (Beetul et al., 2016; Brown et al., 2014; Xu et al., 2017). They
usually inhabit shallow waters and are divided into two main sub-groups (based on size), viz.
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macroalgae (commonly known as seaweed) and microalgae (Aliyu et al., 2021). Marine algae, either
micro or macro, are very significant primary producers in the marine food chain. Moreover,
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macroalgae are composed of multiple organized cells in structures resembling leaves, stems, and
roots, while microalgae are comprised of unicellular microscopic photosynthetic organisms (Aliyu et
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al., 2021). El Gamal (2010) stated that marine algae are potential sources of secondary metabolites in
the aquatic ecosystem. As the name suggests, macroalgae, or seaweed, can be viewed through naked
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eyes, having a length of up to a few meters, are the most common species in the production of
phycocolloids, agar carrageenan, etc.(Deviram et al., 2020). Marine macroalgae can be classified into
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several major groups according to their pigmentation and chemical composition, viz. brown algae
(Phaeophyceae), red algae (Rhodophyceae), and green algae (Chlorophyceae) (El Achaby et al.,
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2018; Xu et al., 2017). As reported by the Food and Agriculture Organization (FAO), by the year
2018, the global industry had already exploited 12 million t/yr of 221 species of macroalgae with a
market size valued at USD 6 billion per year (Ferdouse et al., 2018). Macroalgae, better known as
seaweed, have been majorly utilized (almost 85 % of the total market) as a source of human food,
particularly in Japan, China, and Korea. Other applications of seaweed are mainly for hydrocolloid
production, animal feed, fertilizers, biomass for fuels, and wastewater treatment (Casoni et al., 2020;
Seghetta et al., 2017; Sudhakar et al., 2019; Yuan et al., 2019).
Macroalgae or seaweed are multicellular marine organisms that are found near sea-beds. It is
one of the most abundant sources of polysaccharides, including alginates, agar, fucoidan, agarose,
carrageenan, and ulvan (Wahlström et al., 2020). Agar, for example, is derived from polysaccharides
called agarose, which form the supporting structure of the red algae cell wall. Agarose can form gel
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due to its gelling capacity, making it very useful in skincare, herbal medicine, and film-based
products (Rhim, 2011). Red algae are predominantly diverse in Asian countries. It was found that
Balliopsis prieurii and Sirodotia delicatula from Malaysia have a distant relation to Balliopsis
prieurii from South America, suggesting a cryptic species of algae in South America (Johnston et al.,
2014). Meanwhile, carrageenan is a linear sulfated polysaccharide extracted from red edible
seaweed. Carrageenan is widely exploited based on its hydrophilic and anionic properties. It has been
widely used in the food packaging and pharmaceutical industries as gelling, emulsifying, stabilizing
agents, and base materials for packaging films (Abdul Khalil et al., 2017). Brown algae are
multicellular marine organisms located on intertidal shores at high-temperature latitudes. They
exhibit very effective morphological properties such as plasticity that enable specific responses to

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environmental constraints, including sea currents, exposure to ultraviolet radiation, limited light

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availability, and grazier attacks (Charrier et al., 2012). Brown algae contain mostly alginate, where
alginic acid is a linear copolymer with homopolymeric blocks of (1-4)-linked β-D-mannuronate and
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its C-5 eprimer α-L-guluronate residues, respectively, covalently linked together in different
sequences. The ability of alginates to react with divalent and trivalent cation is widely used in
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alginate film formation. Alginic acid is insoluble in water but swells when placed in water.
Increasing production of macroalgae for growing market demands has developed its
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opportunity to be applied as additional cellulose derivatives. The study of cellulose-based macroalgae

began long ago. The first study on seaweed extraction from green seaweed was first reported in 1885
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(Mihranyan, 2011). In general, macroalgae or seaweeds contain 80 – 90 % water with dry weights
basis, contains carbohydrates in the range of 4 % to 76 %, 1 – 3 % lipids, and 7 – 38 % minerals.
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Meanwhile, their protein content can vary in the vicinity of 10 % to 47 %, with high proportions of
essential amino acids(El-Said and El-Sikaily, 2013). However, the chemical composition of
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microalgae varies according to the species and the maturity of the biomass (Starko et al., 2018;
Wahlström et al., 2020). Cellulose seems to be present in most red algae (Rhodophyta phylum) and
all brown algae (Phaeophyceae class) (Zanchetta et al., 2021). According to Yoon et al. (2010) and
Jeong et al. (2012), red algae, particularly Gelidium amansii possess a high carbohydrate content of
around 75 %, including cellulose and agar. Meanwhile, Albuquerque et al. (2021) reported the
amount of carbohydrate owned by red algae Gracilaria birdie to be almost 73 %, with only 8%
protein content. On the other hand, the content of carbohydrates in the brown algae, including
Saccorhiza polyschides, Sargassum muticum, and Sargassum filipendulla lies in the vicinity of 45 –
52 % (Erbabley and Junianto, 2020; Rodrigues et al., 2015). Whereas, among the green algae, the
cellulose production focused on Ulvophyceae class with species Ulva lactuca, which is found to have
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a 54.3 % content of carbohydrate that is considered high and applicable for the production of
nanocellulose (Kim et al., 2011; Wahlström et al., 2020; Zanchetta et al., 2021).
The cellulose derived from different species of macroalgae was reported to have likenesses to
cotton-based cellulose. The recognition was done based on three different analyses, including (i)
production of D-glucose through hydrolysis, (ii) presence of β-(1,4) linkage via preparation of
cellubiose octa-acetate, and (iii) similar comparison between X-ray peak of algal cellulose and cotton
cellulose (Baghel et al., 2021). The comparable characteristics between these celluloses can be
differentiated through the compositional ratios of two allomorphs. The cellulose produced from
macroalgae is dominated by α-allomorphs (α-cellulose), while the cellulose derived from cotton
sources is dominated by β-allomorph (β-cellulose). The allomorphs possessed by various species of

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macroalgae, including those belonging to green, red, and brown seaweed, predominantly belong to α-

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allomorphs. In addition, the species belongs to the agarophytic and alginophytic groups, which
contain a high content of cellulose and α-cellulose, while the members of the carrageenophyte are
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vice versa. In terms of thermodynamic reactivity, the dominance of α-allomorphs in macroalgae-
based cellulose gives superior properties over other cellulose derived from different sources, like
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woody and plant biomass. The macroalgae-based cellulose also has a 100 times larger surface area
than algal cellulose extruded into a single strand compared to that of plant-based cellulose
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(Phanthong et al., 2018; Zanchetta et al., 2021).

The cellulose content can be differentiated according to their species, which belong to the red,
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green, and brown seaweed groups. According to the study done by Baghel et al. (2021), a total of 19
species of green seaweed possess cellulose content in the range of 1.5 – 34 % of the dry weight, with
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an average of 9.67 %. Meanwhile, the cellulose content for the same 47 species of red seaweed was
in the vicinity of 0.85 – 18 %, with an average of 4.75 %. The cellulose content was calculated based
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on 15 species of brown seaweed and ranged from 2.2 – 10.2 % on a dry weight basis with an average
of 7.88 %. The overall cellulose content for seaweed biomass was reported to be in the range of 0.85
and 34 %. These values are in line with the information available on the cell walls of the algae
according to their groups (Zanchetta et al., 2021). The high cellulose content in green seaweed is
attributed to the main fibrillary layer of polysaccharides comprised of cellulose, hemicellulose, and a
pectin blend. The cell wall of the brown algae is composed of mainly alginate, and fucoidan consists
of a small fraction of cellulose, mainly cellulose microfibrils. The cell wall of red algae generally
contains sulfated galactan consisting of agar and carrageenan, which usually do not represent the
cellulose, explaining their low cellulose content (Allard and Templier, 2000; Domozych, 2016;
Kröger et al., 2018; Zanchetta et al., 2021).
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The development in technology regarding biomass processing has shown great advancement in
facilitating the successful extraction of cellulose from various types of biomass. This innovative
development has provided the opportunity to develop large-scale production of cellulose and its
utilization in a wide range of applications. The presence of cellulose in macroalgae cell walls and
their superior properties enable it to be utilized in diverse extraction strategies. Nanocellulose is one
of the unique and promising natural materials produced from cellulose. However, until today, there
were only a few studies related to the extraction of nanocellulose from marine biomass or seaweed
waste.

2.1. Nanocellulose isolation strategies

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Marine algae, especially macroalgae, have gained wide attention as nanocellulose derivatives

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attributed to their fast growth rate. This growing rate enables us to meet the increasing demand for
nanocellulose production. The high concentration of carbohydrates and low content of natural
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physicochemical barriers in marine algae need no severe chemical treatment to remove the non-
cellulosic component (Chen et al., 2016; Ilyas and Atikah, 2021). Besides, the amount of lignin in
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algae was found to be very low (less than 5 %), according to what was observed in the algal cell
(Chen et al., 2016; Ilyas and Atikah, 2021). This characteristic of algae benefits the pre-treatment
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and hydrolysis processes by easing the elimination of non-cellulosic components. Thus, it yields high
cellulose pulp quality.
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The extraction and isolation of nanocellulose, mainly from marine algae biomass, is not widely
reported compared to other lignocellulosic materials such as plant biomass. Nevertheless, to date,
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marine algae have attracted the attention of both academia and industry due to their unique biological
resources that comprise a high content of proteins and polysaccharides. Currently, researchers and
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manufacturers are working on finding an effective conversion of marine biomass into high-value-
added products, including nanocellulose (Yu et al., 2021). Most of the study dedicated to microalgae
species, including from Ulvophyceae class (green algae) (e.g., Ulva lactuta, Valonia ventricosa,
etc.), Phaeophyceae class (brown algae) (e.g., Crystosphaera jacquinottii, Fucus vesiculosus, etc.),
and Rhodophyta class (red algae) (e.g., Gelidium elegans, etc.) are reported for cellulose and
nanocellulose production (Zanchetta et al., 2021). The most common isolation steps subjected to
marine algae for nanocellulose extraction have been reported. They include (i) the defibrillation
process, for example, alkaline treatment using sodium hydroxide (NaOH) where most parts of
hemicellulose are partially solubilized, and residual waxes, pectin, and natural fats are eliminated,
(ii) the purification of cellulose pulp from other impurities, including oxidative bleaching involving
hydrogen peroxide (H2O2) and sodium chlorite (NaClO2) and (iii) the last step, which is the most
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important in determining the types of nanocellulose produced. The mechanical disintegration process
is employed for the complete defibrillation of cellulose components into individual fibrils to produce
nanofibrillated cellulose (NFC). While nanocrystalline cellulose (NCC) is isolated through acid
hydrolysis using strong mineral acids like hydrochloric acid and sulfuric acid (Ilyas et al., 2017; Ilyas
and Atikah, 2021; Ventura-Cruz and Tecante, 2021; Zanchetta et al., 2021).
The production of NFC generally involves two types of pre-treatment processes, viz. biological
(enzymatic) and classical chemical pre-treatment (TEMPO oxidation) and mechanical pre-treatment.
These processes reduce the particle size, hence increasing the contact surface area and increasing the
effectivity of subsequent treatments (Karimi et al., 2013). The extraction of NFC via chemical pre-
treatment is comprised of two categories, including (i) the modification of cellulose hydroxyl groups

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to carboxyl, sulfonate (Naderi et al., 2017), phosphoryl (Noguchi et al., 2017), and cationic

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ammonium sites (Wågberg et al., 2008) and (ii) opening of cellulose ring and modified other
functional groups by introducing the aldehyde groups. The periodate oxidation has been applied to
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the ring-opening method to cut the C-C bond between the C2 and C3 positions of the cellulose ring
to form aldehyde groups and turn them into alcohol, carboxyl, and sulfite groups, or cationic
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ammonium. Besides, ozonation is also included as one of the cheap methods for carbonyl group
introduction. Meanwhile, mechanical pre-treatment has also been applied in addition to biological
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and chemical pre-treatment, which includes refining, blending, and grinding (Qing et al., 2013;
Zanchetta et al., 2021). The grinding process is very important as it can help to impart uniform
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particle size (Chen et al., 2011). The pre-treated cellulose fiber was then subjected to mechanical
treatments to provide total defibrillation of individual fibers using high shear force processes. The
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defibrillation process includes high-pressure homogenization via homogenizer or microfluidizer,

ultra-fine friction grinding, and refining (Davoudpour et al., 2015; Taheri and Samyn, 2016; Uetani
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and Yano, 2011; Wang et al., 2012). These processes have been commonly used for the delamination
of cellulose bundles for NFC production and have been considered suitable for upscaling.
On the other hand, nanocrystalline cellulose (NCC) was widely reported as being produced via
the acid hydrolysis process. One of the most common chemical processes is to reduce particle size,
increase crystallinity, and improve the lignocellulosic materials’ properties (Kusmono et al., 2020).
However, it is important to determine suitable conditions such as the type of acid, acid concentration,
temperature, and reaction time that contribute to the characteristics of the produced NCC. The
reagents for the hydrolysis process mostly reported in the literature are sulfuric acid (H 2SO4) and
hydrochloric acid (HCl). The acid reagent was responsible for attacking the fibers' amorphous region
and leaving the crystalline part to remain insoluble in acids (Ng et al., 2015). During the process, the
acids led to the esterification of the hydroxyl groups with sulfate ions and introduced a negative
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charge on the surface of the crystals. The charged surface contributes to the anionic stabilization of
the crystal's colloidal suspension resulting from the electric double layer repulsive/attractive forces
(Kargarzadeh et al., 2012). Kusmono et al. (2020) addressed the optimal conditions for the acid
hydrolysis process were 45 ˚C, 30 min, with 58 % sulfuric acid to produce NCC with 91 %
crystallinity and diameters 6.67 nm. However, these conditions might vary according to the fibers
subjected to the treatment. Nonetheless, due to the effectiveness of this treatment to provide high
crystallinity nanocellulose with a stable suspension, this treatment has been considered to be the
main approach to produce NCC as it is also an economical and productive process (Kian et al., 2020;
Ventura-Cruz and Tecante, 2021; Zinge and Kandasubramanian, 2020).
Despite the superior nanocellulose properties produced, the concentrated acid solution imparts

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an unfavorable impact on the environment. But still, from the commercial point of view, the

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manufacturers continue to utilize this conventional method as a production method for NCC (Haldar
and Purkait, 2020). In addition, this method also provides higher yield production of NCC, which
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makes this method considered for industrial commercialization. However, several challenges need to
be taken into accounts, such as high capital investment for reactors, tanks, piping requirements, the
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effect of the effluent produced on the environment, and the operational costs that may increase for
intensive effluent treatments. A similar problem was also faced with nanocellulose production via the
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mechanical process. Both chemical and mechanical processes require high energy for the
disintegration process and a high amount of water for the neutralization process (Ribeiro et al.,
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2019).
To date, enzymatic hydrolysis has been introduced as a processing route to produce
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nanocellulose, promoting a more environmentally friendly and sustainable approach. This method
has utilized an enzyme, which possesses high substrate specificity, targeting specific lignocellulosic
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linkages. This approach offers a better solution than conventional methods as it produces no toxic
residues and can perform under mild conditions, resulting in a less energy-intensive process. Also,
the nanocellulose derived through the enzymatic process has more value in applications due to its
morphologies (Ribeiro et al., 2019; Squinca et al., 2020).

2.2. Marine algae nanocellulose characteristic

Nanocelluloses are unique and promising products derived from natural materials like marine algae.
In general, nanocellulose can be categorized into three main groups based on their structure and
source, viz. nanocrystalline cellulose (NCC), nanofibrillated cellulose (NFC), and bacterial cellulose
(BC) (Zanchetta et al., 2021). The last decades have witnessed increased studies published for
nanocellulose preparation derived from marine algae or seaweed biomass through different
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approaches. The characteristics of the nanocellulose can be varied according to the method used
during the extraction and isolation process. The sources of cellulose are also responsible for
determining the properties of the nanocellulose product. Thus, it is challenging to predict and
recommend the best conditions for nanocellulose production (Baghel et al., 2021). Therefore, many
studies have been done to compare the properties of the nanocellulose produced from different types
of cellulose sources and between other marine algae groups and species.
Nanocellulose imparts a wide spectrum of advantageous physical, chemical, and biological
properties. The outstanding characteristics of nanocellulose are widely reported through physico-
chemical studies of the nanocellulose, such as nanoscale scale, non-toxicity, high surface area, high
aspect ratio and stiffness, crystallinity, interesting mechanical and morphological characteristic, and

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good thermal stability (Rashid et al., 2021; Trache and Thakur, 2020). These characteristics measure

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and built up new opportunities for developing nanocellulose, mainly derived from algae biomass. To
date, there have been extensive studies done on the preparation of nanocellulose from different
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lignocellulosic fibers, including cotton, flax, ramie, sisal, and wood fibers. This nanocellulose
usually becomes the reference to be compared with nanocellulose derived from marine algae. Baghel
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et al. (2021) reported that nanocellulose prepared from seaweed possesses higher crystallinity
indexes (58-99 %) and diameters (ranges between 5 nm and 45 nm) than nanocellulose extracted
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from lignocellulosic biomass. This finding was supported by Yu et al. (2021). The superiority of
nanocellulose prepared from marine algae showed higher cellulose crystallinity compared to that of
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nanocellulose produced from lignocellulosic and agricultural residues. Hua et al. (2015) also found
that the nanocellulose extracted from marine biomass from Cladophorales orders via acid hydrolysis
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gives high crystallinity up to 100 % and also a high specific area.

Meanwhile, Xiang et al. (2016) compare the properties of green seaweed of similar
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Cladophorales order, specifically Chladofora glomerata with bleached eucalyptus pulp. The
cellulose was extracted via enzymatic hydrolysis followed by mechanical nanofibrillation using a
domestic blender at 20000 rpm. The prepared nanofibrils’ were reported to have a 96.5 % crystalline
index. The size of the seaweed was reported to be between 10 – 40 nm, which is smaller than
eucalyptus pulp nanofibrils. The thermal stability of the NFC produced from green seaweed is also
higher compared to the NFC produced from eucalyptus pulp. Besides, in a separate discussion, Van
Hai et al. (2015) had compared the properties of nanocellulose produced from wood, cotton linters,
cattails, and red algae. Among all the nanocellulose tested in the study, red algae showed higher
thermal stability and crystallinity indexes as compared to wood-derived NCC. Later, the researchers
explore the properties of nanocellulose produced from different types of marine algae based on their
groups and species. For example, El Achaby et al. (2018) prepared nanocrystals from red seaweed
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residues using different hydrolysis times viz. 30, 40, and 80 min. The nanocrystals produced with 80
min hydrolysis time showed an optimum yield of 11.18 % nanocellulose with 87 % crystallinity and
a 5.2 ± 2.9 nm diameter. While Liu et al. (2017b) prepared cellulose nanocrystals using kelp waste
with a yield of 52.3 % and 69.4 % crystallinity index.
Other important characteristics of nanocellulose extracted from marine algae include its
thermal stability. According to Kumar et al. (2021), the thermal stability of cellulose fiber is essential
especially when it is used as additive or nanofiller in polymer nanocomposites. The thermal
properties of the nanocellulose can vary according to the types of algae and their method of
extractions. For example, Yu et al. (2021) had compared the characteristics of nanocellulose
extracted from red algae exhibited higher thermal properties. In contrast, nanocellulose produced

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from brown algae, for instance, Lamanaria japonica, presented poor thermal properties. Despite the

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variation in the thermal properties of the nanocellulose derived from marine algae, the thermal
stability of the nanocellulose generally showed superior properties as compared to non-extracted
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algae biomass. The lower thermal resistance of the algae biomass can be caused by the presence of
impurities that cause fast degradation (Paniz et al., 2020). For better comparisons, the characteristics
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of seaweed nanocellulose based on different species and production methods were summarized and
listed in Table 1.
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Table 1: The crystallinity and size of nanocellulose produced from different marine algae species
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Size,
Marine algae Type of Crystallinity T max
Treatments diameter References
species nanocellulose Index (%) ( ℃)
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(nm)
Gelidium NC Chemical 73 17.7-39.9 334 (Chen et al.,
elegans treatments 2016)
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Cladophora NFC Mechanical 92 20-30 - (Hua et al.,

cellulose treatment 2014)
(TEMPO)
Cladohophra NFC Mechanical 96.5 10-40 326 (Xiang et
glomerata and enzymatic al., 2016)

Ulva lactuta NFC Mechanical 63 - 342 (Wahlström

and chemical et al., 2020)
treatments
De-alginate NCC Mechanical 74.5 % - 120- (Feng et al.,
kelp residue and chemical 180 2015)
treatments
Laminaria NCC Chemical 69.4 20-45 240 (Liu et al.,
japonica treatments 2017b)
Gelidiella NCC Mechanical 60 32 ~360 (Singh et
acerosa and chemical al., 2017)
treatments
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Red algae NCC Chemical 81-87 2.3-12.2 215- (El Achaby

waste treatments 330 et al., 2018)

Sargassum NCC Mechanical 58-79 4.8-41 - (Doh et al.,

fluitans, and chemical 2020c)
Laminaria treatments
japonica,
Palmaria
palmate,
Porphyra
umbilicalis,
Ulva lactuta
Laminaria NCC Chemical 85-99 8-21 370- (Doh et al.,
japonica, treatments 480 2020b)
Sargassum

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natans
C. MFC Chemical 71.02 - - (Paniz et al.,

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Jacquinotii treatments 2020)
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Considering the interest received by nanocellulose from research and industry, it is crucial that
in-depth risk evaluation are performed to ensure their safety before commercialization. In general,
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nanocellulose is considered materials with no toxicity and immunogenicity or low cytotoxicity and
immunogenicity that do not contribute to serious environmental hazards and are widely applied in
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biomedical applications. In fact, major studies on this matter were ambiguous, for instance, in vitro
studies to evaluate the inflammation, cytotoxicity, oxidative stress, and genotoxicity (Stoudmann et
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al., 2020). The reason lies in the perspective of the purified materials viz. nanocellulose, either NFCs
or NCCs, there are no significant differences in their cytotoxicity have been reported according to
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production route. However, the different properties of a nanoparticle might dictate its cytotoxicity,
including its dimension, shape, reactivity, and other chemical or physical properties (Michelin et al.,
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2020). Besides, recent studies also revealed that nanocellulose, particularly NCCs might cause an
inflammatory response, mages, cause oxidative stress induction, and be able to enter cells due to its
nature of surface chemistry, which may influence the toxicity in a different manner (Trache et al.,
2020). However, Bacakova et al. (2019) reported that such immunogenicity and cytotoxicity can be
controlled by endowing and electrical charges or modification using specific chemical groups. The
study on the eco-toxicological impact on cellulose nanocrystals (NCC) was done by Kovacs et al.
(2010) through a realistic exposure scenario viz. suspension experiment with relevant dose ranges
based on the potential effluent in the vicinity of an NCC production. This study includes aquatic
organisms from all trophic levels, including bacteria, algae, crustacean, cnidarian, and fish, to
investigate acute lethality upon reproduction growth, morphology, embryo development, and
cytotoxicity. The final results were consequently summarized as “non-concerning.” In a separate
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study, Hua et al. (2015) evaluated the response of monocytes and macrophages to functionalized
Cladophora nanocellulose films in terms of inflammatory response. The carboxylated functionalized
Cladophora nanocellulose films (a-CC) under pro-inflammatory stimuli with its aggregated, aligned
fiber structures caused the more pronounced reduction of TNF–α levels compared to hydroxypropyl
trimethylammonium-substituted CC-film (c-CC) that in turn impart non-aggregated, randomly
oriented fibers. The authors summarized this finding as a successful finding that suggested that these
products of nanocellulose derived marine algae can be applied for specific applications as dressing
for treating chronic wounds.
Nevertheless, the effect of various surface modifications might need further investigation. The
relation between functionalization and toxicity is currently not parallel, as different types of

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modification can show diverging results. On the other hand, findings on the effect of nanocellulose

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on the genotoxicity were reported for NCC, and NFC showed no significant effect. For example,
Catalán et al. (2017) produced nanocrystalline cellulose (NCC) derived from cotton and found no
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genotoxicity towards human bronchial epithelial cells using micronucleus assay to assess
chromosomal damage. Whilst, the genotoxicity of NFC was studied by Pitkänen et al. (2014) on
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Salmonella typhimurium using the bacterial reversion assay also showed no genotoxicity effect.
However, follow-up testing might be needed for the first screening of evaluation for the studied
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materials.
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3. Microbial treatment for nanocellulose extraction

Production of nanocellulose via microbial host as the degrading agent has become increasingly
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popular owing to its environmental-friendly properties. As a result, several research groups have
developed and published processes for nanocellulose bio-production to develop methods for large-
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scale production (Jozala et al., 2016). In general, there are three types of nanocellulose viz. (a)
cellulose nanofibrils (NFC), which are commonly known to be produced through an enzymatic
and/or mechanical isolation process, (b) cellulose nanocrystals (NCC), which can be obtained via
strong acid hydrolysis using hydrochloric acid and sulfuric acid, and (c) bacterial nanocellulose,
which is synthesized exclusively by a family of bacteria known as Gluconoacetobacter xylinus
(Ribeiro et al., 2019). The production of nanocellulose, particularly via mechanical and chemical
means, requires intensive mechanical fibrillation and vigorous chemical reactions, respectively,
causing potential environmental pollution. The nanocellulose production via microbial-based
extraction via enzymatic hydrolysis can help reduce the effects presented by both mentioned
methods. Besides, the combination of enzymatic post-treatment seems to be a better alternative for
nanocellulose isolation (Bian et al., 2020).
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3.1. Enzymatic hydrolysis

In recent years, the ever-growing demand for nanocellulose derived from algae, wood, and non-wood
plant biomass has promoted the exploration of more convenient methods. The microbial extraction
process via enzyme-assisted hydrolysis imparts advantages regarding extraction time, low energy,
and less application of chemical solvents, so it is considered a "green technique." This method is also
considered one of the most potent approaches to extract cellulose from marine algae biomass due to
its high efficiency under mild reaction conditions (Xu et al., 2017). However, enzymes for nano
cellulose production are not widely explored in the literature compared to chemical and mechanical
processes. Nevertheless, there is an effort to evaluate this strategy with other treatments.

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(a)
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(b)

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Fig. 1. Microbial extraction of nanocellulose via enzymatic hydrolysis according to the (a)
breakdown of each process using different micro-organisms and (b) the overall schematic of the
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enzymatic hydrolysis mechanism for nanocellulose production [adapted from Michelin et al., (2020)
from http://creativecommons.org/licenses/by/4.0/]. R: reducing cellulose end; NR: nonreducing
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cellulose end; EG: endogluconases; CBH I: exogluconases removing cellobiose from R end; CBH II:
exogluconases removing cellobiase from NR end; β-G: β-glucosidase; CDH: cellubiose
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dehydrogenases; LPMO: lytic polysaccharides monoxygenases

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In general, enzymatic hydrolysis can be performed using various micro-organisms, including

bacteria, yeast, fungi, etc., as presented in Fig. 1(a). However, the hydrolysis process might vary
according to the micro-organism used. Still, this method has been discussed as an alternative pre-
treatment method to mechanical and chemical pre-treatment approaches. Based on studies, this
process can provide promising alternatives to chemical analysis as it is compatible with biological
processes that can ease the nanostructure yield and increase its thermal stability (Michelin et al.,
2020). The utilization of enzymes in the isolation of nanocellulose can be a complex process,
including removing pectin and hemicellulose and enzyme applications to isolate the cellulose
segment by weakening and disrupting the fiber's cell wall structure (Rodrigues et al., 2015). The
mechanism of nanocellulose production involving enzymes, namely cellulase as the specific class of
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enzyme. It has been commonly used to catalyze the hydrolyzation reaction of cellulose that breaks it
into smaller polymer segments, as shown in Fig. 1(b).
Due to its promising technique, this method has been considered a sustainable route for
nanocellulose production. It is safe for the environment because it does not produce any toxic by-
products. This advantage brings a growing interest in the use of enzymes for nanocellulose
production. However, there are limited, or no studies on the production of nanocellulose derived
from marine algae via enzymatic hydrolysis except for the production of fermented sugars for
biofuels (Khammee et al., 2021; Kostas et al., 2020). Therefore, the overview of the production of
nanocellulose from other plant-based cellulose via enzymatic hydrolysis using different micro-
organisms was discussed here to represent the potential of marine algae biomass as a suitable and

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susceptible raw material to yield nanocellulose through enzymatic analysis.

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Many commercially available enzymes have been used to assist the extraction process of
compounds such as viscozyme, cellucast,-p termamyl, ultraflo, carragenase, agarase,
amyloglucosidase, xylanase, kokuzyme, protamex, neutrase, flavourzyme, and alcalase (Wijesinghe
and Jeon, 2012; Xu et al., 2017). Nevertheless, one class of enzymes corresponds to the majority of
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these cases known as “cellulase.” Cellulase is a specific class of enzymes that catalyzes the cellulose
hydrolyzation process and produces simple sugars (Michelin et al., 2020). Cellulase hydrolyzed
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bonds either in the form of endo- or exo-acting within the cellulose molecules or at the reducing or
non-reducing ends (Jeoh et al., 2017). Enzymes can be categorized into three group’s viz. (i)
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endoglucanases (endocellulolases), (ii) cellobiohydrolases (exoglucanases), and (iii) β-glucosidases

(cellobiases) (Barcelos et al., 2015; Choudhury et al., 2020; Jaiswal et al., 2021; Nechyporchuk et al.,
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2016; Ribeiro et al., 2019; Singh et al., 2021). Whilst, these enzymes can be produced by various
types of micro-organisms, including anaerobic bacteria (Clostridium, Ruminococcus,
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Caldicellulosiruptor, Acetivibrio, Butyrivibrio) and aerobic bacteria (Cellulomonas, Thermobifida,

Cytophaga, Sporocythophaga), actinomycetes (streptomyces) filamentous fungi (Bulgaria,
Chaetomium Helotium, Coriolus, Phanerochaete, Poria, Schizophyllum, Serpula), plants (Fragaria),
and animals (mollusks, insects) (Barcelos et al., 2015; Lynd et al., 2002; Palomer et al., 2004).
Cellulase has been known to be applied in the hydrolysis process of lignocellulosic biomass to obtain
sugar liquor by converting the cellulose substrate to glucose enzymatically. The produced glucose is
then used as a building block for a wide range of molecules, from fuels to polymers (Barcelos et al.,
2015). The cellulase enzyme has diversified applications in various industries like agriculture,
bioconversion (ethanol from cellulose), detergent (cleaning with fiber damage), fermentation
(development of wine), food (clarification of fruit juices), pulp and paper (Vigneshwaran and
Satyamurthy, 2016).
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3.2. Nanocellulose production via enzymatic hydrolysis

Several studies have been reported the nanocellulose production through enzyme catalyze process
using various types of lignocellulosic fibers, such as wheat straw (Espinosa et al., 2019), bagasse
pulp (Yassin et al., 2019), orange peel (Hideno et al., 2014), sugarcane straw (De Aguiar et al.,
2020), the cotton pulp (Chen et al., 2019), and bleached hardwood (Dai et al., 2018). The condition
of the process varied depending on the source of cellulose materials and the type of targeted
nanocellulose being produced. According to Michelin et al. (2020), this process comprises two main
steps: the extraction of the cellulose and the isolation of the nanocellulose. The process is initiated by
reducing the size of the materials through mechanical (e.g., grinding, milling) and physicochemical

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(e.g., auto-hydrolysis, alkali, bleaching) pretreatment. The pre-treatment process also purifies and

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extracts cellulose by eliminating hemicellulose, lignin, and extractive components. The extracted
cellulose then continued in a subsequent process involving enzymatic modification of the purified
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cellulose.
Earlier, the cellulose extraction involved the pre-treatment subjected to lignocellulosic to break
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the interconnecting structure of its composition. The surface area of the polymer was exposed and
facilitated the effectiveness of the enzymatic treatment. The common pre-treatment applied at this
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stage is the mechanical process followed by the washing process. The second step is the primary
process of nanocellulose production involving enzymatic hydrolysis. At this stage, the pre-treated
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cellulose substrate was solubilized in the buffer solution with an enzymatic cocktail and broken into
smaller cellulose polymers. The endoglucanases are the enzymes used for hydrolysis of the
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amorphous region and preserve the crystalline cellulose domain accordingly. This process occurs in
relatively mild conditions, pH 4 to 7 and temperatures between 45 ˚C to 50 ˚C. The mixing time can
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be in the range of 1 h to 72 h according to the types and conditions of lignocellulosic materials and
the type of micro-organism used. The third step is the process of imparting the homogenization
consistency of the nanocellulose. The method includes mechanical means such as the
homogenization process (Henriksson et al., 2007; Jaiswal et al., 2021; Martelli-Tosi et al., 2016;
Michelin et al., 2020; Ribeiro et al., 2019).
The mechanism of enzymatic hydrolysis, as depicted in Fig. 1 (b), to transform cellulose into
monosaccharides can be achieved through a chemical process carried out by a special group of
cellulolytic enzymes, namely cellulase. As mentioned in the previous section, cellulase is divided
into three groups’ viz. endogluconases (EG), exogluconases, and cellobiohydrolases (CBH). As
reported in the literature, the hydrolysis process of cellulose into glucose requires the synergistic
action of at least two of the three enzymes. The first enzyme helps in cleaving random internal β-1-4-
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glycosidic bonds of the cellulose chain through enzyme endogluconases (EG). The cleavage of the
cellulose chain typically occurs in the amorphous region, resulting in a new cellulose chain end.
Then, cellobiohydrolases (CBH) or exogluconases act on the cellulose chains to form cellubiose
units. Finally, the class of cellulase refers to β-glucosidase hydrolyzes the cellobiose into glucose.
Overall, the endogluconases are the focus of interest throughout the mechanism. Their action is
focused on the amorphous region of the cellulose and breaking it into smaller-length polymers. This
synergistic effect can be combined with other mechanical and acid treatments for the complimentary
process to produce nanocellulose.
The literature on the use of enzymatic hydrolysis for nanocellulose production is minimal—
some researchers are focusing on the preparation of nanocellulose from common lignocellulosic

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fibers. The process is also usually assisted with different pre-treatment steps. These pre-treatments

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are needed to remove the non-cellulosic components, including lignin and hemicellulose. Martelli-
Tosi et al. (2016) discussed the importance of pre-treatment upon fibers undergo enzymatic
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hydrolysis using soybean straw. The authors found the pre-treatment help to increase the yield of
nanocellulose produced with better thermal stability. A similar study was also done before by
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Henriksson et al. (2007), which reported a combined treatment involving enzyme endoglucanases
and mechanical shearing of wood fiber pulps to yield nanofibers with a high aspect ratio and
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molecular mass compared to those produced from acidic treatment. This method imparts a better
effect on the environment, free from toxic and corrosive chemicals commonly applied in acidic
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treatment. The application of enzymatic hydrolysis to assist dilute acid pre-treatment for fermentable
sugar production of marine macro-algae Nizimuddin zanardini was also reported by Yazdani et al.
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(2011). The maximum yield of glucose from macroalgae was reported through enzymatic
saccharification using cellulase, and β-gluicosidase indicate the effectiveness of enzymatic
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hydrolysis to degrade the cellulose polymer.

As discussed earlier, enzymatic hydrolysis can be performed using different types of enzymes
generated by different types of micro-organisms (bacteria, fungi, and yeast). This micro-organism
can generate different types of enzymes that can be utilized for the hydrolysis process for
nanocellulose production. Although the literature on the function of different kinds of enzymes has
not yet been reported on marine algae biomass, their function as catalysts for nanocellulose
production for other types of cellulose derivatives has been discussed and presented in the upcoming
section.

3.2.1. Enzymatic hydrolysis using bacteria

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Microbial cellulose can be produced via enzymatic hydrolysis using various types of bacteria. These
bacteria produce different enzymes or combinations of different enzymes, which are preferentially
used to decompose cellulose serially or synergistically and convert it into monosaccharides (sugars).
Most bacteria produce enzymes such as Bacillus subtills, Pseudomonas fluorescens, E.coli, and
Serratia marcescens., Cellvibri Cellulomonas, Bacillus, Pseudomonas sp. and Micrococcus,
Streptomyces thermoviolaceus, Streptomyces sp. Streptomyces roseiscleroticus, Streptomyces galbus,
Bacillus circulans (Polizeli et al., 2005). Bacteria Caldicellulosiruptor bescii (thermophilic bacteria)
has been reported to produce multifunction enzymes that can be utilized to produce nanocellulose
from lignocellulosic biomass. It imparts some advantages, including reducing the cost of
bioconversion, decreasing the contamination probability of microbes, improving the mixing rate, and

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enhancing kinetics. Meanwhile, in another study, Pseudomonas fluorescens was used to clean lignin

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biomass by producing cellulolytic enzymes, where hemicellulose is degraded into monosaccharides
by cleaving carbohydrates into sugars (Ghosh et al., 2019).
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The isolation of nanocellulose through degradation from bacterial enzyme cocktail due to
hydrolytic activities from Xanthomonas axonopodis pv. citri strain 306 (IBSBF 1594) was also
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reported using citrus waste biomass. The production process and the effect of before and after
enzymatic digestion was depicted in Fig. 2. The morphological analysis showed the conversion of the
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citrus-waste biomass from fiber to nanocellulose (Fig. 2b-e). The successful conversion indicated that
enzymatic hydrolysis using this bacterial enzyme cocktail was very efficient. In addition, an
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improvement of 7 % in NFCs content and a 13 % in crystallinity index have been attained through
enzymatic hydrolysis. Further, the mechanical treatment of NFCs produces almost 55 % crystallinity
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with a 10 nm average diameter (Mariño et al., 2015).

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Fig. 2. The production of nanocellulose from citrus waste biomass via enzymatic hydrolysis is
depicted in (a) process flowchart, including the pre-treatment and production of bacterial enzymes
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used for the hydrolysis process. The SEM micrographs of (b) citrus biomass treated with NaOH
during delignification, (b) pretreated citrus biomass after Xanthomonas axonopodis pv. citri
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enzymatic activity, (c) fibers and fiber bundles of citrus biomass after enzymatic action, and (d) the
FESEM micrographs of nanocellulose fibers after sonication process [adapted from Mariño et al.
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(2015) from http://creativecommons.org/licenses/by/4.0/]

3.2.2. Enzymatic hydrolysis using yeast

Yeasts are single-cell organisms that convert carbohydrates into alcohol through fermentation. To
date, yeast cellulases have been utilized in enzymatic hydrolysis through fermentation. Yeast can
withstand high temperatures up to 70 ˚C and can produce thermostable cellulase, and can be used at a
broader range of pH, which is needed for the fermentation process (Giese et al., 2011). Research has
been conducted, and 82 samples of yeast strains from various sources (tree bark, soil, and insect
frass) were selected and optimized for enzymatic activities. Identification was made on both solid and
liquid mediums. Out of 45 yeast strains, three strains (UBU-SK6, UBU-JK8, and UBU-JK9/1)
showed the highest cellulose-degrading capabilities in solid medium. They are then grown on a liquid
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medium with 1 % yeast extract, varying carboxymethyl cellulose (0.5 -1.5 %) for 168 h at 30 °C
under shaking (150 rpm). Later these three yeast strains were identified molecularly and showed
close similarities to yeasts of Candida sp. (Candida sp. 05-7-186T, Candida easanensis, and Candida
sp. ST-390 respectively) (Thongekkaew and Kongsanthia, 2016). The first type of cellulases was
produced by Trichosporon laibachii MG270406-1A14 strain. In this study, 23 yeast strains were
isolated from natural sources (carbon and nitrogen) from decaying wood and attine ants for
hydrolases. Out of all, two yeast strains of Trichosporon laibachii MG270406-1A14 strain confirmed
higher cellulase concentration. Cellulase activities, β-glucosidase, and Xylanase were found in
Pichia, Trichosporon, Cryptococcus, and Debaryomyces genera. Physiological and biochemical
characteristics indicated that the yeast strain has the potential for cellulolytic enzyme and can be

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utilized for enzymatic hydrolysis of lignocellulosic biomass (Giese et al., 2017).

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The nanocellulose isolation via fermentation has been reported by Tsukamoto et al. (2013).
Isolation of nanocellulose from citrus processing waste from oranges (CPWO) using three different
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strains Saccharomyces cerevisiae and two Candida sp. namely Candida parapsilosis strains IFM
48375 and NRRL Y-12969 (ATCC 22019), has been studied. The CPWO was enzymatically
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converted into fermentable sugars, and finally, nanocellulose (2.5 % g/g dry CPWO) and nanofibers
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(0.5 % g/g dry CPWO). Nanocellulose and nanofibers were successfully isolated from solid leavings
from enzymatically hydrolyzed and fermented CPWO. Božič et al. (2014) reported the application of
yeast to improve the biocompatibility of the NFC using commercial hexokinase (Saccharomyces
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cerevisiae). Touijer et al. (2019) choose Trichosporon genus out of 30 selected isolates of yeasts to
expresses the characteristics of hydrolyzing cellulose fiber, including carboxymethyl cellulose
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(CMC) and filter paper (FP). The results confirmed that the obtained cellulase was highly
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thermostable under an operating temperature range of 55 °C to 70 °C with an ample pH of 5. It can

be potentially used for the enzymatic saccharification of soluble (cellulose fiber, filter paper (FP))
and insoluble carboxymethyl cellulose (CMC) substrates.

3.2.3. Enzymatic hydrolysis using fungi and other micro-organisms

Fungi are one of the micro-organisms which produce cellulase enzymes used for the production of
nanocellulose. Fungal enzyme reactions play a prominent role in keeping the environment. Cellulase
enzymes generally degrade cellulose under controlled hydrolysis by fungi. In the case of aerobic
fungi, cellulases are produced (endoglucanases (EC 3.2.1.4), cellobiohydrolase (EC 3.2.1.91), and
cellobiase (β-glucosidase, EC 3.2.1.91)) as a multi-component enzyme. It acts synergistically in
hydrolyzing cellulose and producing glucose without mechanical treatment. However, filamentous
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fungi are the main commercial source. Most studied fungi belong to genes Aspergillus sp. and
Trichoderma sp. for enzymatic treatment of cellulosic biomass for production of nanocellulose. The
produced nanocellulose surface can be modified by using xylanase through enzymatic treatment. The
enzymatic process also removes residues of lignin and hemicellulose. Xylanase was produced by
fungi, particularly from the genera Aspergillus and Trichoderma fungi (Polizeli et al., 2005).
Nanocrystalline cellulose was successfully isolated by a combination of anaerobic micro-
organisms from the MCC derived from cotton fibers. The treatment was performed enzymatically,
followed by a differential centrifugation technique (for purification), which resulted in spherical
nanocellulose forming 12.3 % in 7 days. Atomic force microscopy confirmed bimodal size
distribution (43 ± 13 and 119 ± 9 nm) in spherical nanocellulose without any surface modification

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and less energy consumption (Satyamurthy and Vigneshwaran, 2013). A similar approach was

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applied by Hayashi et al., using an exoglucanase (CBH I) to shorten the MCC from algal cellulose of
Cladophora sp., it resulted increased in crystallinity by degrading amorphous characteristics
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(Hayashi et al., 2005). Hasanin et al., (2018) reported a comparable finding using the fungus
Humicola Fuscoatra Egyptia X4 to produce nanocellulose from agricultural waste. The results
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showed that nanocellulose produced was of high quality with high crystallinity, high surface area,
high surface area, and high stability having dimensions (93 ± 5.6 nm long and 12 ± 4.7 nm wide).
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In a separate discussion, the utilization of fungi-based enzymatic hydrolysis was reported to

enhance the hydrophobicity of nanocellulose. The fact is that cellulose can absorb water when
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exposed to it; therefore, surface modification is needed to maintain its internal morphology. Garcia-
Ubasart et al. (2012) utilized laccase from T. versicolor to improve the hydrophobicity of the
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nanocellulose and increase the contact angle up to 80˚. Another method for increasing hydrophobicity
is acetylation. Researchers utilized acetylation with acyl donors in enzymatic treatment by using
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fungal strains. Nanocellulose was acetylated by acetic anhydride in the enzymatic hydrolysis process
using lipase from Aspergillus niger species. Acyl modification of hydroxyethylcellulose has also
been reported in the presence of β-galactosidase from fungi Aspergillus oryzae. Transesterification
using acylation agents, such as vinyl acrylate and vinyl propionate, can increase the hydrophobicity
of nanocellulose (Thomas et al., 2018).

4. Current market demand and ecological concerns of marine algae nanocellulose

The production of nanocellulose using locally available, biodegradable, and inexpensive organic
waste for conventional nanomaterial substitution in many applications is intended to contribute
towards sustainability and reduce environmental impact. Its utilization depends on its chemical
composition, components, yield generated, and superior mechanical and thermal performance, which
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have become important aspects in product development (Ventura-Cruz and Tecante, 2021).
Nowadays, nanocellulose has been recognized as the main material used on a large scale by many
major industries, including pulp and paper production, the packaging industry, energy-based
applications, and environmental remediation (Dhali et al., 2021). This is proved by the data of the
global nanocellulose market that reported worth USD 284.7 million in 2018 and projected to increase
to USD 661.3 million by 2023 (MarketsandMarkets.com, 2020). The application of nanocellulose
was focused more on the utilization of NFC by accounting for the largest share of the nanocellulose
market in 2018, followed by NCC. In general, considering the price of the raw materials (~ USD$ 0.5
per kg) and the production cost, the price of NFC was estimated to cost between USD$ 7 – US$ 12
per kg of dried material (Dhali et al., 2021). Meanwhile, the cost of NCC ranging from USD$ 4 –

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USD$10 per kg (de Assis et al., 2017).

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Other than crops, the source of cellulose materials, marine algae could be the potential source
of cellulose and nanocellulose derivatives used in various applications, as proved in many research
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reports. Previously, marine algae were known for their broad applications as food, fodder,
pisciculture, fertilizer, etc. Due to their rich source of carbohydrates, fats, protein, and vitamins A, B,
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C, and E, as well as minerals including iron, potassium, magnesium, calcium, and zinc, have made
them a food ingredient in many major countries, such as Ireland, Scotland, Sweden, Japan, and
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Korea. Global algae production is categorized based on type, source, form, application, and region.
The algae harvesting was approximately 4000-5000 metric tons every year, resembling more of a
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crop plant, resulting in the evolution of a strong economic crop (Veluchamy and Palaniswamy, 2020).
The annual growth rate of algae production was estimated to increase by 4.2 % between 2018 and
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2025 due to the high demand for natural products. Therefore, there is an immense opportunity to
extract marine algae-based cellulose to add variety to the nanocellulose class for various applications.
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The cellulose yield generated from marine algae was also reported to be comparable to that of
lignocellulosic biomass, in the range of 27-45 %, which has become feasible and economical for
nanocellulose production (Baghel et al., 2021).
Marine algae are one of the biomass wastes represented by the high value of biopolymer
resources. Nanocellulose is extracted from marine algae biomass through different techniques, either
used to develop composites or offered to support a wide range of applications and other new
emerging areas. As a material derived from a renewable energy source, the development of
nanocellulose through high value-added products can save energy and solve some intractable
environmental problems while promoting economic development (Yu et al., 2021). For example, the
biodegradability properties of nanocellulose have made them eligible to be actively used in the
preparation of polymers, films, composites, hydrogels, etc. The biodegradability of nanocellulose has
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been a hot topic for researchers. For instance, the application of NCC in the film has shown an
increment of biodegradation properties according to ISO 17556:2003 (E) standard (Bagde and
Nadanathangam, 2019). Nanocellulose based products were also tested for their degradation behavior
through a soil burial test. Whilst, Funabashi et al. (2009) have suggested several other evaluations to
analyze the biodegradation of the nanocellulose based polymer composites according to ISO 14855-1
and 14855-2, which referred to compost biodegradability test for pilot scale and laboratory scale,
respectively. Other standards considered important for evaluating the biodegradability of materials
include ASTM D5511-18 and ASTM 6400, referred to as standard test methods for anaerobic
biodegradability and standard specification for solid material biodegradation, respectively (Zinge and
Kandasubramanian, 2020).

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The global industry has shown its interest in developing more efficient and cleaner production

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approaches that sustainably help product manufacturers reduce negative environmental, economic,
and social impacts. Therefore, the focus on more environmentally friendly and cost-effective methods
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for producing nanocellulose has become very important. Instead of being economically feasible and
effective, their effect on environmental safety also becomes a crucial factor that needs to be focused
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on. For example, the utilization of chemicals for bleaching and the delignification process is very
effective in eliminating lignin but very harmful to the environment. As an alternative, hydrogen
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peroxide under controlled conditions was used to replace conventional pre-treatment techniques and
was more effective in imparting better nanocellulose products. Despite the technique's effectiveness,
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some limitations need to be overcome, viz. the excessive water usage for purification of isolated
materials from the chemical by-products resulting from the extraction process. Excessive water usage
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can be considered a disadvantage as it can affect the process cost and energy consumption.
The search for the diversity of nanocellulose production has found new alternatives that can
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reduce the total cost of the process and the environmental impacts related to the production methods.
Microbial extraction through enzymatic hydrolysis offers several advantages, including mild reaction
conditions, less or no production of hazardous waste, reduction of water consumption, and the
possibility of using the secondary stream of fermentable sugars released in the hydrolysis process
(Michelin et al., 2020). This approach has also been explored to minimize the energy requirement
needed for mechanical treatment to produce NFC and reduce or eliminate the use of chemicals such
as sulfuric acid and TEMPO for NCC production. However, the disadvantages of this method are the
high cost of the enzymes, and the procedure is slower than conventional methods. However,
considering the significant economic returns on improved quality products, such as higher purity,
easily functionalized nanostructures, with better thermal stability, specific areas, and aspect ratios,
support this method's use.
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5. Application of marine algae nanocomposites

Advance development in biopolymer research has shown the superiority of biopolymer materials in
various applications, particularly for biopolymers produced by micro-organisms (Jozala et al., 2016).
Algae biopolymer is a promising biomaterial that has rarely been explored despite its unique
properties as a functional material. The demand for cellulose-derived materials is on a steady
increase due to their abundance, low cost, and biodegradability, including improved mechanical
properties and thermal stability. Recently, marine algae have gained remarkable interest due to the
inherent abundance of cellulose and the benefit of the absence of lignin, making algae biomass a
suitable precursor for nanocellulose. These nanocellulose showed high potential as new types of

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reinforcing filler in the field of composite materials. The features, including preparation of the

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nanocellulose, the dispersion, and interfacial compatibility between nanocellulose and the polymer
matrix, impart superiority to the resulted nanocomposites, determining the feasibility of
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nanocellulose composites in the industry. High mechanical and thermal stability, chemical inertness,
and tailored morphological, physical, chemical, electrical, thermal, and optical properties, barrier
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properties, antimicrobial effects, biocompatibility with no or low toxicity, and low immunogenicity
play a major role in determining the breakthrough point and direction of utilization of nanocellulose
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composites in various fields of industries. Besides, they are relatively low-cost materials with high
availability and renewability. Therefore, these benefits possessed by nanocellulose make it a
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potential candidate for a wide range of industrial, technological, and biomedical applications, namely
packaging, biomedical and pharmaceutical products, including cell culture and drug delivery, as well
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as other parts of applications, including pollution control, filtration and ultrafiltration, water
treatment, 3D printing, and electronics (Aliyu et al., 2021; Bacakova et al., 2019; Fang et al., 2019).
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5.1. Packaging
The concept of bio-economy through the synthesis of biomaterials has created a sustainable approach
towards applying biodegradable materials as functional bio-product for food packaging to enhance
an effective circular economy (Jafarzadeh et al., 2020). Cellulose derived from algae is an attractive
prospect for sheet coating and binders. Fibers of cellulosic composition have a high degree of
polymerization, including high modulus and tensile strength for selecting and fabricating
biocomposites. Rajesh Kumar Dora et al. (2020) have developed a biopolymer film derived from
algae bioproduct with adequate physical and mechanical properties for food packaging applications.
The produced algal coating film exhibited high compressive and tensile stresses, including high
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elastic modulus. The effective mechanical properties could result from the high surface coating
thickness and variation in the modulus of substrate and coating.
Meanwhile, de Oliveira et al. (2019) have produced bioactive aerogels biopolymers from algae
biomass for food packaging applications via a physical crosslinking technique. The authors
suggested that the high relative humidity of the aerogel could be attributed to the high water sorption
and the impact of the high hydrophilic cellulose content. The potential of the produced aerogel to
interact with water relative to the hydrophilic cellulosic composition. This characteristic can be
attributed to the availability of hydroxyl groups that interact with water through the pores'
interconnectivity that allows the inward movement of water through the matrix.
Sun et al. (2014) have developed a biodegradable biomass packaging material from Eichhornia

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Crassipes. The pulverization of the algae biomass resulted in the conversion of macromolecule fibers

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into microfibers, which allows the interaction of hydroxyl group active sites for physical absorption.
The addition of adhesives to the pulverized precursor reinforced the biopolymer further to improve
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the bio-packaging material. It has been reported that the valorization of algae can be utilized as food
packaging materials. In their investigation, biomass derived from the Gelidium sesquipedale seaweed
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after agar extraction was used as the precursor for producing cellulosic fractions used to generate
film from the suspension. The authors attributed the surface roughness to the amorphous matrix
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attributed to the residual agar, which embedded the compacted structure of the cellulose fibers.
Moghaddas Kia et al. (2018) produced a biodegradable food packaging film using Zedo gum/sodium
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caseinate (ZG/NaCs) incorporated with microalgae (Spirulina platensis) as an antioxidant

compound. Authors attributed the increase in water vapor permeability to the film's hydrophilic
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behavior, which has the affinity for water and decreased tortuosity of the path of vapor diffusion
through the film.
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The enhancement of the biopolymer-based macroalgae was also studied by Abdul Khalil et al.
(2018) and Hasan et al. (2019) using microbial calcium carbonate (MB-CaCO3) for the preparation
of plasticulture packaging film (Fig. 3). The red seaweed macroalgae (Kappaphycus alvarezii) has
been prepared with the addition of 0.15% MB-CaCO3 showed the brighter film with better water
barrier and biodegradability compared to that incorporated with commercial CaCO3.
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Fig. 3. The fabrication of red seaweed Kappaphycus alverazii incorporated with microbial and
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commercial calcium carbonate (MB-CaCO3 and C-CaCO3) for plasticulture packaging film.
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In a separate discussion, Abdollahi et al. (2013) found the properties of alginate biopolymer
can be enhanced by the addition of cellulose nanoparticles (CN) with the highest content of CN up to
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5 %. Meanwhile, Doh (2020a) utilized the nanocellulose production from marine algae biomass
Sargassum fluitans and Laminaria japonica applied to alginate nanocomposites film. Instead of
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improving the film's mechanical and water vapor permeability properties, nanocellulose help
prolonged the biodegradation process, which imparts high potential to be applied for a novel
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biomaterial in the packaging system. These findings on applying biodegradable polymers from algae
have a promising prospect for biocomposite formulation under various conditions for food packaging
as an alternative to utilization of synthetic packaging materials that have resulted in severe
environmental pollution over the years.

5.2. Biomedical
Recently, macroalgae and their derivatives have been widely proposed for biomedical applications.
Marine algae biopolymers have been reported as a replacement for many synthetic polymers due to
their biocompatibility and nontoxic properties (Abdul Khalil et al., 2020). The development of
cellulose and nanocellulose based algae biomass has attracted a lot of attention for many
applications, including tissue engineering and drug delivery systems. Its unique physical,
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mechanical, chemical, and biological properties offer many advantages over synthetic polymers. Fig.
4 illustrates the potential of marine algae biomass to be applied in one of the biomedical applications,
viz. tissue engineering for skin graft treatment.

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Fig. 4. The schematic representation of marine algae biomass for tissue engineering applications
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Besides that, algae also demonstrated vast potential based on their thermal stability and
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improved mechanical properties as a prospective nanofiller for reinforcement application in

composites, which can serve valuable purposes in developing scaffolds for bone and tissue
engineering. For instance, macroalgae-based nanocellulose has already been explored as a scaffold
for human dermal fibroblast cells (Beaumont et al., 2021). Hua et al. (2014) had compared the effect
of microfibrillated nanocellulose (MFC) and algal nanocellulose extracted from Cladophora sp. on
the material's cytocompatibility. The addition of the algae nanocellulose was then found to promote
fibroblast adhesion and support high cell viability. While, in other separate discussions, pure
nanocellulose algal extracted from Cladophora has been studied by Liu et al. (2017a), showing the
various impurities found in algal-based nanocellulose, particularly Cladophora sp. were acceptable
for future biomedical applications. The application of macroalgae in the biomedical is also employed
in the form of a hydrogel. In their study, Kavitha et al. (2019) employed hydrogel formulated from
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the sodium salt of alginate synthesized from T. conoides for targeted biomedical applications. Two
hydrogel formulations of alginate-based biomaterials were synthesized to form extracted
alginate/collagen microfilm for wound healing and extracted alginate microbead for anticancer drugs
that have been proved for biomedical applications in pharmacology.
In addition, Liu et al. (2017a) utilized macroalgae biomass by extracting
sulfated polysaccharides from Ascophyllum nodosum brown seaweed, which exhibited antitumor
activity in sarcoma-180 tumor-contained mice. The injection of the B16 cells was attributed to anti-
metastatic activity in the mouse model due to the effect of the incorporation of ascophyllan. It was
revealed that the trans-well assay suppressed the invasion potential of B16 cells. The authors
concluded that the number of lung surface metastatic nodules in ascophyllan-treated mice was

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reduced significantly compared to the untreated control mice. The B16 cells could have been

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influenced by the activation of the immune system by ascophyllan as a result of in vivo anti-
metastatic activity on B16 melanoma cells. In a related study, Ale et al. (2011) have reported that
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brown seaweeds possess essential bioactive properties that influence inhibitory effects on tumor
cells. To this effect, the authors investigated the significance of fucose-derived sulfated
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polysaccharides from Sargassum sp. on the growth of melanoma B16 cells and cell viability. It was
suggested that fucose-derived sulfated polysaccharides Sargassum sp induced the proliferation of
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melanoma B 16 cells while the analysis of cell viability inhibited the proliferation of the melanoma
cells. Thus, it is suggestive that the presence of sulfated galactofucans polysaccharides derived from
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Sargassum sp. possibly could exert bioactive effects on skin cancer cells. Meanwhile, Rocha et al.
(2007) have reported in vitro screening from extracts derived from Lobophora variegata on the
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impact of cytotoxic activity on a cultured human melanoma cancer cell with the use of
sulphorhodamine B assay. Authors revealed that semi-purified fraction of Lobophora variegate
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inhibited the growth of melonama cells while Spatoglossum schroederi extract from crude acetone
could not inhibit the growth of melanoma cells.
In recent studies, marine algae have also been recognized as potential antiviral agents to
prevent recent Corona Virus Disease-2019 (COVID-19). The sulfated polysaccharide in marine
algae is known to have various antiviral activities against a broad range of enveloped and non-
enveloped viruses, becoming a potential source of antiviral drugs for pharmaceutical development.
Hans et al. (2021) stated the mechanism of these sulfated polysaccharides in inhibiting the different
stages of the viral infection process inside the host, which involves blocking the initial entry of the
virus by modulating the immune response of the host cell. Previously, the potential of other plant
extracts to combat the combat against COVID-19 had been studied. For example, Shree et al. (2020)
found the potential of phytochemicals medicinal plants, for instance, Ashwagandha (Withania
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somnifera), Giloy (Tinospora cordifolia), and Tulsi (Ocimum sanctum), as an inhibitor against
SARS-Cov-2 main protease. However, the potential of algae-based and plant-based compounds
possesses merits and demerits. Even though no clear health risk has been identified in the literature,
the requirement for additional information on chemical, biological potency, bioavailability, toxicity,
and associated mechanisms of sulfated polysaccharides is needed for them to be applied in the
pharmaceutical sector.

5.3. Environmental and wastewater treatment

The environmental system and natural water bodies are affected by pollution from wastewater due to
the harmful chemicals, dyes, and heavy metals it contains. The application of algae biomass as an

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adsorbent for the removal of recalcitrant pollutants is economical due to the abundance and

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sustainability resulting from the efficiency of the bio-material as functional adsorbents in wastewater
treatment systems (Tokay and Akpınar, 2021). It is widely reported the use of algae as a biosorbent
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due to the scarce requirement for nutrients, high sorption capacity, plentiful availability, high surface
area to volume ratio, less volume of sludge, and the potential for metal regeneration and recovery.
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Furthermore, the components in marine algae, such as sulfated polysaccharides, showed the potential
of algae-based biomaterials to be applied as absorbent materials. The extraction of the chemical
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components via enzymatic hydrolysis can be considered both an economical and eco-friendly
solution for wastewater treatment via the mechanisms shown in Fig. 5 (Kanamarlapudi et al., 2018).
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Fig. 5. The schematic illustration of marine algae biomass biomaterials with adsorbent capability for
wastewater treatment applications.

Developing cost-effective and highly efficient treatment technologies for pollutants removal
before their discharge to the environment and receiving water is significant. Some of the treatment
methods include Fenton oxidation, advanced oxidation, zonation, coagulation, and adsorption.
Adsorption is a widely recognized method due to the low cost of treatment, ease and abundance of
adsorbents, and highly efficient removal of pollutants from effluents. Many adsorbent materials have
been reported to remove various types of contaminants, including adsorbent minerals, biosorbents,
activated carbon, and nanomaterials (Barquilha and Braga, 2021). Meanwhile, algae biomass has

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remarkable application in wastewater treatment using adsorption, especially for removing emerging

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organic contaminants. Besides, most types of algae possess sulfated polysaccharides, such as brown
algae, and red algae contain high amounts of carboxyl groups that are usually involved in the
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biosorption of metals (Kanamarlapudi et al., 2018). Mustapha and Halimoon, (2015) reported algal
biomass capability to remove the metal ions from contaminated water resources about 15.3 % - 84.6
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% higher than other microbial biosorbent. To date, algal-based biosorbents have been commercially
available; for example, AlgaSorb by Biorecovery Inc. has algal biomass immobilized in silica gel. In
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a pilot study, two columns in series with different biosorbents of algae, namely AlgaSorb 624 and
AlgaSorb 620, were used to remove mercury from groundwater (Barkley, 1991).
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In separate studies, marine algae can also be cultured and produced valuable biomass for
industrial applications. For example, the experiment conducted by Gramegna et al. (2020)
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demonstrated the culturing of microalgae from Chlorrella sp. combined with Clamydomonas
reinhardtii strain to be used for dairy wastewater remediation and production of lipids and/or
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recombinant enzymes. The co-cultivation of microalgae and microbes was observed to be able to
reduce the pollution load in dairy waste, decreasing nitrogen and phosphorus levels in wastewater
discharge limits and remarkably lowering COD and protein nitrogen. In addition, the production of a
thermostable β-glocosidase by transplastomic Clamydomonas reinhardtii grown in dairy waste
showed the potential of sustainable biorefinery waste remediation with the production of
recombinant enzymes and microalgae.
Nanocellulose has been recognized as an excellent biomaterial adsorbent for wastewater
treatment due to its adsorption affinity towards organic pollutants (Shak et al., 2018). To date,
nanocellulose derived from green algae has been used to resolve issues concerning water pollution in
coastal areas. Despite being a good bio-adsorbent material, nanocellulose also has the potential to be
applied in membrane filtration technology. It is most widely used in water treatment techniques to
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efficiently remove the pollutants such as organic and inorganic particles, heavy metal ions, dyes,
viruses, organic solvent, pesticides and herbicides, and other dangerous substances. Besides,
nanocellulose based microalgae also benefited from protecting the environment from the spreading
viruses such as influenza. Influenza viruses are well adapted in the aquatic environment and able to
replicate and transmit via water-land birds. This method was proven due to its high efficiency,
chemical stability, and low environmental impact (Cao et al., 2013; Olivera et al., 2016; Wang et al.,
2013, 2014). The utilization of the nanocellulose-based membrane also relies on its well-defined
pore size and tailored surface chemistry to remove the contaminants from the water. For example,
Metreveli et al. (2014) had employed nanocellulose extracted from macroalgae species Cladophora
to form a membrane filter paper or membrane. It was found no infectious swine influenza virus

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(SIV) with a diameter size of 80 – 120 nm is unable to pass through and contaminate the triplicate

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96-well plates in the filter, which matches the retention efficiency of the industrial filters. The
development of a composite membrane made from nanocellulose was also utilized for water
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treatment applications. Xiong et al. (2017) constructed an all-natural, transparent, mechanically
robust, and nanoporous biocomposites membrane from the mixture of silk fibroin and cellulose
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nanofiber (NFC). A peculiar “shish kebab” nanostructure with a periodic arrangement of silk fibroin
on the heterogeneous surface of the NFC was reported. This peculiar “shish kebab” SF-NFC
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structure had enabled the membrane to possess a nonporous, transparent, and mechanically robust
intertwined network. It showed high water flux, excellent nanofiltration efficiency for various
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organic molecules, and better absorption for heavy metal ions.

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5.4 Dye removal

The removal of dyes from receiving water is very important to ensure environmental safety. Colored
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dye wastewater imparts environmental degradation, and the by-product is carcinogenic. Removing
this contaminant down to safe levels requires high energy and unsustainable techniques, including
oxidation, biodegradation, photo-catalyst, precipitation, and reverse osmosis, and is expensive.
Consequently, the combination of biosorption and nanotechnology offers alternatives to remediating
this chemical contamination(Norrrahim et al., 2021). In addition, this approach was preferred based
on its high effectiveness and being economically viable (Choudhury et al., 2020). Nanocellulose has
caught the attention of water treatment engineers, particularly for dye removal. It has worked as a
high-performance adsorbent due to its suitable surface properties, such as high surface area,
hydrophilic surface chemistry, and uniform distribution of functional groups (Norrrahim et al., 2021;
Trache et al., 2020).
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The adsorption properties possessed by nanocellulose are the main features for its
consideration as a contaminant removal material. Adsorption is a simple method that provides a
cheaper and user-friendly approach; no by-products are formed at the end of the process. The
contaminants molecules adhere to the absorbent surface by physical forces like van der Waals forces
and electrostatic attraction or through covalent bonding between adsorbate molecules and the
absorbent surface (Rasalingam et al., 2014). The utilization of nanocellulose as an adsorbent material
has been reported to impart better performance concerning adsorption capacity for dye molecules.
The reason lies in several factors, such as the ability of nanocellulose to surface functionalization,
better binding affinity with other materials, ability to be used multiple times to adsorb and desorb
contaminants, renewable and biodegradable, high surface area, high mechanical properties, and good

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surface tension, which favors the wetting of nanocellulose and its stability in water (Islam et al.,

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2013; Mo et al., 2019; Voisin et al., 2017). The hydroxyl groups -OH on the cellulose backbone of
nanocellulose also introduced the desired functionality and produced highly effective flocculants for
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decontamination purposes.
The nanocellulose isolated from macroalgae was seen to be utilized in dye removal in water
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treatment applications. According to Mohamed et al. (2017), the mechanism of absorption and
separation can be comprised into four categories viz. physical, chemical, biological and acoustical,
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radiation, and electrical processes. The application of nanocellulose can be regenerated by desorption
of dyes using suitable eluent like ethanol, dilute acid solution, etc. (Choudhury et al., 2020). The fact
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that nanocellulose possesses an abundance of –OH on its segment, making it excellent water and
other polar liquids absorbents. Nanocellulose can also be functionalized through sulfuric acid
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hydrolysis (SO3+), carboxylated groups (COO-), and amines (-NH2) groups ionically or covalently.
This functionalized nanocellulose can be used for selective removal of targeted contaminants
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(Mohamed et al., 2017a).

The efficacy of nanocellulose adsorbents against dyes removal was reported as high as
nanocomposites and more than functionalized nanocellulose (Ma et al., 2017; Melo et al., 2018; Qiao
et al., 2015). Sustainable materials like nanocellulose is an ideal material for contaminants removal
in wastewater treatment. Mohammed et al. (2015) have built a new generation of recyclable
adsorbents with combined nanocellulose and alginate biopolymer materials with improved
adsorption capacity. The nanocomposites in hydrogel beads were evaluated using methylene blue
(MB) batch adsorption in an aqueous solution. Through this study, the authors confirmed that the
adsorption process through thermodynamic analysis is spontaneous and exothermic. The maximum
adsorption capacity of the CNC-ALG hydrogel beads is comparable to activated carbon through the
equilibrium adsorption data, with a value of 256.41 mg/g. The removal efficacy of MB also remained
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almost 97 % after five adsorption-desorption cycles, attributed to the effectiveness of CNC-ALG

hydrogel beads for the removal of organic dyes from wastewater.
Meanwhile, a biopolymer-based aerogel synthesized from crosslinking bi-functional hairy
nanocrystalline cellulose and carboxylated chitosan through a Schiff base reaction was developed for
environmental remediation application. This bio-aerogel was examined in a model using methylene
blue (MB) cationic dye. The equilibrium adsorption data was found to be approximately 785 mg/g
with equilibrium constant K in the vicinity of 0.0089 at room temperature. Through this finding, the
bio-aerogel, the combination of nanocellulose with modified chitosan, exhibits the highest removal
capacity compared to other reusable adsorbents prepared from biopolymers. Besides, this material
also showed effective adsorption-regeneration cycles with excellent reusability with adsorption

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capacity remained constant over a wide pH range (e.g., pH >7).

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6. Challenges and future perspective of marine algae nanocellulose
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To date, algae biopolymers have been utilized for various applications, such as packaging,
environmental and biomedical. Consequently, few species have been recognized to reach an
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industrial and commercial scale. Marine macroalgae have been reported to have been widely utilized
for industrial applications compared to microalgae as they are readily accessible and easy to harvest
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and low in cost of production (Beetul et al., 2016). Another potential application of marine algae is
the derivative for the production of cellulose. The extraction of cellulose from algal biomass into
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functional materials is essential for reducing the effect of eutrophication on the aquatic ecosystem.
However, its sustainability has not been fully explored, especially as reinforcement materials as
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fillers in composites. This could partly be attributed to the availability of biomass, even though quite
a substantial number of marine algae exhibit potential for large-scale cultivation. Still, factors such as
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pH conditions and the availability of light as an energy source for converting CO 2 into biomass via
photosynthesis determine the growth rate of microalgae and biomass. These factors create various
limitations to determine the identification of types and characteristics of marine algae that require
expertise for the identification process and are time-consuming (Beetul et al., 2016).
Nonetheless, despite the drawbacks, the isolation of cellulose from various types of macroalgae
biomass for desired functional materials creates the potential for a novel approach towards high-
value-added products and applications. In addition, there are wide opportunities for cellulose derived
from macroalgae to be developed with the growing marine biomass global production (Baghel et al.,
2021). However, other technical challenges and cost issues need to be addressed, especially when
large-scale production and application are possible (Yu et al., 2021). Based on the detailed analysis
done, cellulose extracted from macroalgae possesses similar properties to that derived from the
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lignocellulosic fiber. Although it has a lower cellulose content, its lower percentage of lignin
component eases the extraction process and enables lower production costs. Besides, most
macroalgae species' cellulose yield was also found to be comparable to that of lignocellulosic
biomass (~ 27 – 45 %). Thus, efforts are needed to utilize the cellulose products derived from
macroalgae. These factors offer better economic value for cellulose as a marine biomass co-product
and create better feasibility in the industries (Baghel et al., 2021).
Besides that, the high demand for cellulose-derived marine algae has become the focus of
future research to prepare advanced and high-value products such as nanocellulose. To date, (. One
solution to achieve better yields focused on pre-treatment improvement during the extraction of the
nanocellulose (Ribeiro et al., 2019). However, comparative studies of the best extraction for specific

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algae species are still lacking. The production of nanocellulose derived from marine algae has been

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reported via conventional processes, including chemical pre-treatment (TEMPO oxidation) and
mechanical pre-treatment (high pressure). Despite using conventional types of extraction, biological
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pre-treatment, such as enzyme-based extraction, offers a solution to financially overcome the lower
yield through integration with other industries, including bio-refineries, food, and juice processes.
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The escalating numbers of studies on the production of nanocellulose via enzyme treatments indicate
the potential of this method. Various types of micro-organisms are employed subjected to enzyme
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treatments. This method is a perfect pathway to give properties that meet the desired properties for
many valuable applications, particularly in the form of reinforced polymer composites.
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As nanocellulose applications are mostly related to the impact of human health and
environmental purification, such as in biomedical, pharmaceutical, food packaging, and
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environmental and wastewater treatment applications, the safety concern of the produced
nanocellulose has become very important. The production of "green materials" derived from natural
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fibers has become an interest in various industrial areas. However, proper investigation needs to be
done to analyze the impact that nanocellulose could have on human health when extensively exposed
to the consumer of the end products (Thomas et al., 2020). The data reported by Endes et al. (2016)
in acute exposure scenarios upon the structure-activity relationship of nanocellulose indicate that
they did not impart greater risk to human health and environment than other nanomaterials in similar
potential applications such as carbon nanotubes.
In Malaysia, the National Institute of Occupational Safety and Health (NIOSH) has assigned a
recommended exposure limit for respirable bulk cellulose to be 5 mg/m3. However, the exposure
limit to nanocellulose is yet to be established due to limited experimental data on the exposure of
nanocellulose to/within in vitro/in vivo systems in environmental conditions that closely mimic
realistic industrial/environments (Ventura et al., 2018). Nonetheless, some effort has been made to
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address the environmental and safety aspects of nanocellulose to ensure commercial applications.
Life cycle risk assessment is one of the methods to identify the potential exposure scenarios, evaluate
the toxicity, and highlight the data needs and gaps to reduce the uncertainty about nanocellulose
safety (Shatkin and Kim, 2015). However, studies on long-term exposures with realistic
concentrations are still lacking, and the carcinogenicity, neurotoxicity, and genotoxic effects of
nanocellulose need to be evaluated (Stoudmann et al., 2020). Therefore, considering the vast interest
received by nanocellulose based products and the rapid development of novel applications, in-depth
analyses must be performed to ensure the safety aspects of the materials before their
commercialization. There are many attempts at various surface modifications on the surface of the
nanocellulose, which might affect the toxicity properties of the materials. Therefore, further

of
investigation into the properties of the material’s surface would be necessary.

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7. Conclusions -p
Macroalgae biomass has been considered a unique biological resource. Thus, converting the algae-
based waste into a high-value-added product has become a significant focus for academia and
re
industry. Several algae species have been identified as an excellent source for nanocellulose
preparation. The nanocellulose production via enzymatic treatments highlights the potential as an
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environmentally friendly pathway that meets the requirements of advanced applications. The
development of nanocellulose via microbial treatment proved the feasibility of these materials to
na

solve intractable environmental problems while promoting economic growth. The potential of
nanocellulose derived from marine algae showed clear potential to be applied in diverse advanced
ur

applications.
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Acknowledgment

The researchers would like to thank the collaboration between Universitas Syiah Kuala, Banda
Aceh, Indonesia, and Universiti Sains Malaysia, Penang, Malaysia, that has made this review
manuscript possible.

Funding

This research was funded by the Ministry of Higher Education Malaysia grant number FRGS-
MRSA 203/PTEKIND/6711837.

References
 Journal Pre-proof

Abdollahi, M., Alboofetileh, M., Behrooz, R., Rezaei, M., Miraki, R., 2013. Reducing water
sensitivity of alginate bio-nanocomposite film using cellulose nanoparticles. Int. J. Biol.
Macromol. 54, 166–173. https://doi.org/10.1016/j.ijbiomac.2012.12.016
Abdul Khalil, H.P.S., Chong, E.W.N., Owolabi, F.A.T., Asniza, M., Tye, Y.Y., Tajarudin, H.A.,
Paridah, M.T., Rizal, S., 2018. Microbial-induced CaCO3 filled seaweed-based film for green
plasticulture application. J. Clean. Prod. 199, 150–163.
https://doi.org/10.1016/j.jclepro.2018.07.111
Abdul Khalil, H.P.S., Jummaat, F., Yahya, E.B., Olaiya, N.G., Adnan, A.S., Abdat, M., Nasir,
N.A.M., Halim, A.S., Seeta Uthaya Kumar, U., Bairwan, R., Suriani, A.B., 2020. A review on
micro- to nanocellulose biopolymer scaffold forming for tissue engineering applications.

of
Polymers (Basel). 12, 1–36. https://doi.org/10.3390/POLYM12092043

ro
Abdul Khalil, H.P.S., Saurabh, C.K., Tye, Y.Y., Lai, T.K., Easa, A.M., Rosamah, E., Fazita, M.R.N.,
Syakir, M.I., Adnan, A.S., Fizree, H.M., Aprilia, N.A.S., Banerjee, A., 2017. Seaweed based
-p
sustainable films and composites for food and pharmaceutical applications: A review. Renew.
Sustain. Energy Rev. 77, 353–362. https://doi.org/10.1016/j.rser.2017.04.025
re
Ahankari, S.S., Subhedar, A.R., Bhadauria, S.S., Dufresne, A., 2021. Nanocellulose in food
packaging: A review. Carbohydr. Polym. https://doi.org/10.1016/j.carbpol.2020.117479
lP

Albuquerque, J.C.S., Araújo, M.L.H., Rocha, M.V.P., de Souza, B.W.S., de Castro, G.M.C.,
Cordeiro, E.M.S., Silva, J. de S., Benevides, N.M.B., 2021. Acid hydrolysis conditions for the
na

production of fine chemicals from Gracilaria birdiae alga biomass. Algal Res. 53.
https://doi.org/10.1016/j.algal.2020.102139
ur

Ale, M.T., Maruyama, H., Tamauchi, H., Mikkelsen, J.D., Meyer, A.S., 2011. Fucose-containing
sulfated polysaccharides from brown seaweeds inhibit proliferation of melanoma cells and
Jo

induce apoptosis by activation of caspase-3 in vitro. Mar. Drugs 9.

https://doi.org/10.3390/md9122605
Aliyu, A., Lee, J.G.M., Harvey, A.P., 2021. Microalgae for biofuels via thermochemical conversion
processes: A review of cultivation, harvesting and drying processes, and the associated
opportunities for integrated production. Bioresour. Technol. Reports 14, 100676.
https://doi.org/10.1016/j.biteb.2021.100676
Allard, B., Templier, J., 2000. Comparison of neutral lipid profile of various trilaminar outer cell
wall (TLS)-containing microalgae with emphasis on algaenan occurrence. Phytochemistry 54.
https://doi.org/10.1016/S0031-9422(00)00135-7
Bacakova, L., Pajorova, J., Bacakova, M., Skogberg, A., Kallio, P., Kolarova, K., Svorcik, V., 2019.
Versatile application of nanocellulose: From industry to skin tissue engineering and wound
 Journal Pre-proof

healing. Nanomaterials 9. https://doi.org/10.3390/nano9020164

Bagde, P., Nadanathangam, V., 2019. Mechanical, antibacterial and biodegradable properties of
starch film containing bacteriocin immobilized crystalline nanocellulose. Carbohydr. Polym.
222. https://doi.org/10.1016/j.carbpol.2019.115021
Baghel, R.S., Reddy, C.R.K., Singh, R.P., 2021. Seaweed-based cellulose: Applications, and future
perspectives. Carbohydr. Polym. 267, 118241. https://doi.org/10.1016/j.carbpol.2021.118241
Barcelos, C.A., Rocha, V.A., Groposo, C., de Castro, A.M., Perreira Jr, N., 2015. Enzymes and
accessory proteins involved in the hydrolysis of lignocellulosic biomass for bioethanol
production, in: Nascimento, R. (Ed.), Mycology: Current and Future Developments. Bentham
Science Publisher Ltd., Rio de Janeiro, Brazil, pp. 23–56.

of
https://doi.org/10.2174/9781681080741115010005

ro
Barkley, N.P., 1991. Extraction of mercury from groundwater using immobilized algae. J. Air Waste
Manag. Assoc. 41. https://doi.org/10.1080/10473289.1991.10466935
-p
Barquilha, C.E.R., Braga, M.C.B., 2021. Adsorption of organic and inorganic pollutants onto
biochars: challenges, operating conditions, and mechanisms. Bioresour. Technol. Reports 15,
re
100728. https://doi.org/10.1016/j.biteb.2021.100728
Bastante, C.C., Silva, N.H.C.S., Cardoso, L.C., Serrano, C.M., Martínez de la Ossa, E.J., Freire,
lP

C.S.R., Vilela, C., 2021. Biobased films of nanocellulose and mango leaf extract for active food
packaging: Supercritical impregnation versus solvent casting. Food Hydrocoll. 117.
na

https://doi.org/10.1016/j.foodhyd.2021.106709
Beaumont, M., Tran, R., Vera, G., Niedrist, D., Rousset, A., Pierre, R., Shastri, V.P., Forget, A.,
ur

2021. Hydrogel-forming algae polysaccharides: from seaweed to biomedical applications.

Biomacromolecules. https://doi.org/10.1021/acs.biomac.0c01406
Jo

Beetul, K., Gopeechund, A., Kaullysing, D., Mattan-Moorgawa, S., Puchooa, D., Bhagooli, R., 2016.
Challenges and opportunities in the present era of marine algal applications, in: Algae -
Organisms for Imminent Biotechnology. InTech. https://doi.org/10.5772/63272
Bian, H., Chen, L., Dong, M., Fu, Y., Wang, R., Zhou, X., Wang, X., Xu, J., Dai, H., 2020. Cleaner
production of lignocellulosic nanofibrils: Potential of mixed enzymatic treatment. J. Clean.
Prod. 270. https://doi.org/10.1016/j.jclepro.2020.122506
Božič, M., Liu, P., Mathew, A.P., Kokol, V., 2014. Enzymatic phosphorylation of cellulose
nanofibers to new highly-ions adsorbing, flame-retardant and hydroxyapatite-growth induced
natural nanoparticles. Cellulose 21. https://doi.org/10.1007/s10570-014-0281-8
Brown, E.M., Allsopp, P.J., Magee, P.J., Gill, C.I., Nitecki, S., Strain, C.R., Mcsorley, E.M., 2014.
Seaweed and human health. Nutr. Rev. 72, 205–216. https://doi.org/10.1111/nure.12091
 Journal Pre-proof

Cao, X., Huang, M., Ding, B., Yu, J., Sun, G., 2013. Robust polyacrylonitrile nanofibrous membrane
reinforced with jute cellulose nanowhiskers for water purification. Desalination 316.
https://doi.org/10.1016/j.desal.2013.01.031
Casoni, A.I., Ramos, F.D., Estrada, V., Diaz, M.S., 2020. Sustainable and economic analysis of
marine macroalgae based chemicals production - Process design and optimization. J. Clean.
Prod. 276. https://doi.org/10.1016/j.jclepro.2020.122792
Catalán, J., Rydman, E., Aimonen, K., Hannukainen, K.S., Suhonen, S., Vanhala, E., Moreno, C.,
Meyer, V., Perez, D. da S., Sneck, A., Forsström, U., Højgaard, C., Willemoes, M., Winther,
J.R., Vogel, U., Wolff, H., Alenius, H., Savolainen, K.M., Norppa, H., 2017. Genotoxic and
inflammatory effects of nanofibrillated cellulose in murine lungs. Mutagenesis 32.

of
https://doi.org/10.1093/mutage/gew035

ro
Charrier, B., Le Bail, A., De Reviers, B., 2012. Plant Proteus: Brown algal morphological plasticity
and underlying developmental mechanisms.
-p Trends Plant Sci.
https://doi.org/10.1016/j.tplants.2012.03.003
Chen, W., Yu, H., Liu, Y., Hai, Y., Zhang, M., Chen, P., 2011. Isolation and characterization of
re
cellulose nanofibers from four plant cellulose fibers using a chemical-ultrasonic process.
Cellulose 18, 433–442. https://doi.org/10.1007/s10570-011-9497-z
lP

Chen, X.Q., Pang, G.X., Shen, W.H., Tong, X., Jia, M.Y., 2019. Preparation and characterization of
the ribbon-like cellulose nanocrystals by the cellulase enzymolysis of cotton pulp fibers.
na

Carbohydr. Polym. 207. https://doi.org/10.1016/j.carbpol.2018.12.042

Chen, Y.W., Lee, H.V., Juan, J.C., Phang, S.M., 2016. Production of new cellulose nanomaterial
ur

from red algae marine biomass Gelidium elegans. Carbohydr. Polym. 151, 1210–1219.
https://doi.org/10.1016/j.carbpol.2016.06.083
Jo

Choudhury, R.R., Kumar, S., Jaydevsinh, S., 2020. Potential of bioinspired cellulose nanomaterials
and nanocomposite membranes thereof for water treatment and fuel cell applications. Cellulose
27, 6719–6746. https://doi.org/10.1007/s10570-020-03253-z
Claro, F.C., Jordão, C., de Viveiros, B.M., Isaka, L.J.E., Villanova Junior, J.A., Magalhães, W.L.E.,
2020. Low cost membrane of wood nanocellulose obtained by mechanical defibrillation for
potential applications as wound dressing. Cellulose 27. https://doi.org/10.1007/s10570-020-
03129-2
Dai, J., Chae, M., Beyene, D., Danumah, C., Tosto, F., Bressler, D.C., 2018. Co-production of
cellulose nanocrystals and fermentable sugars assisted by endoglucanase treatment of wood
pulp. Materials (Basel). 11. https://doi.org/10.3390/ma11091645
Davoudpour, Y., Hossain, M.S., Abdul Khalil, H.P.S., Mohamad Haafiz, M.K., Mohd Ishak, Z.A.,
 Journal Pre-proof

Hassan, A., Sarker, Z.I., 2015. Optimization of high pressure homogenization parameters for the
isolation of cellulosic nanofibers using response surface methodology. Ind. Crops Prod. 74.
https://doi.org/10.1016/j.indcrop.2015.05.029
De Aguiar, J., Bondancia, T.J., Claro, P.I.C., Mattoso, L.H.C., Farinas, C.S., Marconcini, J.M., 2020.
Enzymatic deconstruction of sugarcane bagasse and straw to obtain cellulose nanomaterials.
ACS Sustain. Chem. Eng. 8, 2287–2299. https://doi.org/10.1021/acssuschemeng.9b06806
de Assis, C.A., Houtman, C., Phillips, R., Bilek, E.M.T., Rojas, O.J., Pal, L., Peresin, M.S., Jameel,
H., Gonzalez, R., 2017. Conversion economics of forest biomaterials: risk and financial analysis
of CNC manufacturing. Biofuels, Bioprod. Biorefining 11. https://doi.org/10.1002/bbb.1782
de Oliveira, J.P., Bruni, G.P., Fabra, M.J., da Rosa Zavareze, E., López-Rubio, A., Martínez-Sanz,

of
M., 2019. Development of food packaging bioactive aerogels through the valorization of

ro
Gelidium sesquipedale seaweed. Food Hydrocoll. 89.
https://doi.org/10.1016/j.foodhyd.2018.10.047 -p
Deviram, G., Mathimani, T., Anto, S., Ahamed, T.S., Ananth, D.A., Pugazhendhi, A., 2020.
Applications of microalgal and cyanobacterial biomass on a way to safe, cleaner and a
re
sustainable environment. J. Clean. Prod. https://doi.org/10.1016/j.jclepro.2019.119770
Dhali, K., Ghasemlou, M., Daver, F., Cass, P., Adhikari, B., 2021. A review of nanocellulose as a
lP

new material towards environmental sustainability. Sci. Total Environ. 775, 145871.
https://doi.org/10.1016/j.scitotenv.2021.145871
na

Dias, O.A.T., Konar, S., Leão, A.L., Yang, W., Tjong, J., Sain, M., 2020. Current State of
Applications of nanocellulose in flexible energy and electronic devices. Front. Chem.
ur

https://doi.org/10.3389/fchem.2020.00420
Doh, H., 2020a. Development of seaweed biodegradable nanocomposite films development of
Jo

seaweed biodegradable nanocomposite films reinforced with cellulose nanocrystals for food
packaging reinforced with cellulose nanocrystals for food packaging. Clemson University.
Doh, H., Dunno, K.D., Whiteside, W.S., 2020b. Preparation of novel seaweed nanocomposite film
from brown seaweeds Laminaria japonica and Sargassum natans. Food Hydrocoll. 105,
105744. https://doi.org/10.1016/j.foodhyd.2020.105744
Doh, H., Lee, M.H., Whiteside, W.S., 2020c. Physicochemical characteristics of cellulose
nanocrystals isolated from seaweed biomass. Food Hydrocoll. 102.
https://doi.org/10.1016/j.foodhyd.2019.105542
Domozych, D.S., 2016. Biosynthesis of the cell walls of the algae, in: Borowitzka, M., Beardall, J.,
Raven, J. (Eds.), The Physiology of Microalgae. Springer International Publishing, Cham, pp.
47–63. https://doi.org/10.1007/978-3-319-24945-2_2
 Journal Pre-proof

Dufresne, A., 2013. Nanocellulose: A new ageless bionanomaterial. Mater. Today 16, 220–227.
https://doi.org/10.1016/j.mattod.2013.06.004
Ebadi, Z., Ghaisari, H., Tajeddin, B., Shekarforoush, S.S., 2021. Production and evaluation of the
chemical and mechanical properties of nanocellulose and nanowood starch-based biodegradable
films potential candidates for moisture absorbers for food packaging. Food Sci. Nutr. 9.
https://doi.org/10.1002/fsn3.2194
El-Said, G.F., El-Sikaily, A., 2013. Chemical composition of some seaweed from mediterranean sea
coast, Egypt. Environ. Monit. Assess. 185, 6089–6099. https://doi.org/10.1007/s10661-012-
3009-y
El Achaby, M., Kassab, Z., Aboulkas, A., Gaillard, C., Barakat, A., 2018. Reuse of red algae waste

of
for the production of cellulose nanocrystals and its application in polymer nanocomposites. Int.

ro
J. Biol. Macromol. 106, 681–691. https://doi.org/10.1016/j.ijbiomac.2017.08.067
El Gamal, A.A., 2010. Biological importance
-p of marine algae. Saudi Pharm. J.
https://doi.org/10.1016/j.jsps.2009.12.001
Erbabley, N.Y.G.F., Junianto, 2020. Chemical characteristics and phytochemicals of the brown alga
re
sargassum filipendulla from kelanit waters of south east maluku. Egypt. J. Aquat. Biol. Fish.
24. https://doi.org/10.21608/ejabf.2020.105542
lP

Espinosa, E., Rol, F., Bras, J., Rodríguez, A., 2019. Production of lignocellulose nanofibers from
wheat straw by different fibrillation methods. Comparison of its viability in cardboard recycling
na

process. J. Clean. Prod. 239. https://doi.org/10.1016/j.jclepro.2019.118083

Fang, Z., Hou, G., Chen, C., Hu, L., 2019. Nanocellulose-based films and their emerging
ur

applications. Curr. Opin. Solid State Mater. Sci. https://doi.org/10.1016/j.cossms.2019.07.003

Feng, X., Meng, X., Zhao, J., Miao, M., Shi, L., Zhang, S., Fang, J., 2015. Extraction and preparation
Jo

of cellulose nanocrystals from dealginate kelp residue: Structures and morphological

characterization. Cellulose 22, 1763–1772. https://doi.org/10.1007/s10570-015-0617-z
Ferdouse, F., Lovstad Holdt, S., Smith, R., Murua, P., Yang, Z., 2018. The global status of seaweed
production, trade and utilization, Globefish Research Programme. Rome.
Fritz, C., Jeuck, B., Salas, C., Gonzalez, R., Jameel, H., Rojas, O.J., 2015. Nanocellulose and
proteins: Exploiting their interactions for production, immobilization, and synthesis of
biocompatible materials. Adv. Polym. Sci. 271. https://doi.org/10.1007/12_2015_322
Funabashi, M., Ninomiya, F., Kunioka, M., 2009. Biodegradability evaluation of polymers by ISO
14855-2. Int. J. Mol. Sci. https://doi.org/10.3390/ijms10083635
Garcia-Ubasart, J., Colom, J.F., Vila, C., Hernández, N.G., Blanca Roncero, M., Vidal, T., 2012. A
new procedure for the hydrophobization of cellulose fibre using laccase and a hydrophobic
 Journal Pre-proof

phenolic compound. Bioresour. Technol. 112. https://doi.org/10.1016/j.biortech.2012.02.075

Ghosh, T., Ngo, T.D., Kumar, A., Ayranci, C., Tang, T., 2019. Cleaning carbohydrate impurities
from lignin using: Pseudomonas fluorescens. Green Chem. 21.
https://doi.org/10.1039/c8gc03341b
Giese, E.C., Cadete, R.M., Pierozzi, M., Philippini, R.R., Martiniano, S.E., Pagnocca, F.C., Rosa,
C.A., Da Silva, S.S., 2011. Cellulases production by new yeast isolates from Brazilian
biodiversity. Curr. Opin. Biotechnol. 22. https://doi.org/10.1016/j.copbio.2011.05.491
Giese, E.C., Dussán, K.J., Pierozzi, M., Chandel, A.K., Pagnocca, F.C., Da Silva, S.S., 2017.
Cellulase production by Trichosporon laibachii. Orbital 9.
https://doi.org/10.17807/orbital.v9i4.1024

of
Gramegna, G., Scortica, A., Scafati, V., Ferella, F., Gurrieri, L., Giovannoni, M., Bassi, R., Sparla,

ro
F., Mattei, B., Benedetti, M., 2020. Exploring the potential of microalgae in the recycling of
dairy wastes. Bioresour. Technol.
-p Reports 12, 100604.
https://doi.org/10.1016/j.biteb.2020.100604
Hafid, H.S., Omar, F.N., Zhu, J., Wakisaka, M., 2021. Enhanced crystallinity and thermal properties
re
of cellulose from rice husk using acid hydrolysis treatment. Carbohydr. Polym. 260, 117789.
https://doi.org/10.1016/j.carbpol.2021.117789
lP

Haldar, D., Purkait, M.K., 2020. Micro and nanocrystalline cellulose derivatives of lignocellulosic
biomass: A review on synthesis, applications and advancements. Carbohydr. Polym. 250,
na

116937. https://doi.org/10.1016/j.carbpol.2020.116937
Hans, N., Malik, A., Naik, S., 2021. Antiviral activity of sulfated polysaccharides from marine algae
ur

and its application in combating COVID-19: Mini review. Bioresour. Technol. Reports 13,
100623. https://doi.org/10.1016/j.biteb.2020.100623
Jo

Hasan, M., Chong, E.W.N., Jafarzadeh, S., Paridah, M.T., Gopakumar, D.A., Tajarudin, H.A.,
Thomas, S., Khalil, H.P.S.A., 2019. Enhancement in the physico-mechanical functions of
seaweed biopolymer film via embedding fillers for plasticulture application: A comparison with
conventional biodegradable mulch film. Polymers (Basel).
https://doi.org/10.3390/polym11020210
Hasanin, M.S., Mostafa, A.M., Mwafy, E.A., Darwesh, O.M., 2018. Eco-friendly cellulose nano
fibers via first reported egyptian Humicola fuscoatra egyptia x4: isolation and characterization.
Environ. Nanotechnology, Monit. Manag. 10. https://doi.org/10.1016/j.enmm.2018.10.004
Hayashi, N., Kondo, T., Ishihara, M., 2005. Enzymatically produced nano-ordered short elements
containing cellulose Iβ crystalline domains. Carbohydr. Polym. 61.
https://doi.org/10.1016/j.carbpol.2005.04.018
 Journal Pre-proof

Henriksson, M., Henriksson, G., Berglund, L.A., Lindström, T., 2007. An environmentally friendly
method for enzyme-assisted preparation of microfibrillated cellulose (MFC) nanofibers. Eur.
Polym. J. 43. https://doi.org/10.1016/j.eurpolymj.2007.05.038
Hideno, A., Abe, K., Yano, H., 2014. Preparation using pectinase and characterization of nanofibers
from orange peel waste in juice factories. J. Food Sci. 79. https://doi.org/10.1111/1750-
3841.12471
Hsu, H.H., Zhong, W., 2019. Nanocellulose-based conductive membranes for free-standing
supercapacitors: A review. Membranes (Basel). 9. https://doi.org/10.3390/membranes9060074
Hua, K., Carlsson, D.O., Ålander, E., Lindström, T., Strømme, M., Mihranyan, A., Ferraz, N., 2014.
Translational study between structure and biological response of nanocellulose from wood and

of
green algae. RSC Adv. 4, 2892–2903. https://doi.org/10.1039/c3ra45553j

ro
Hua, K., Strømme, M., Mihranyan, A., Ferraz, N., 2015. Nanocellulose from green algae modulates
the in vitro inflammatory response of monocytes/macrophages. Cellulose 22, 3673–3688.
-p
https://doi.org/10.1007/s10570-015-0772-2
Hubbe, M.A., Pruszynski, P., 2020. Greaseproof paper products: A review emphasizing ecofriendly
re
approaches. BioResources 15. https://doi.org/10.15376/biores.15.1.1978-2004
Ilyas, R.A., Atikah, M.S.N., 2021. Production of nanocellulose from sustainable algae marine
lP

biomass. Semin. Adv. Bio- Miner. based Nat. Fibre Compos.

Ilyas, R.A., Sapuan, S.M., Ishak, M.R., Zainudin, E.S., 2017. Effect of delignification on the
na

physical, thermal, chemical, and structural properties of sugar palm fibre. BioResources 12,
8734–8754. https://doi.org/10.15376/biores.12.4.8734-8754
ur

Irawan, A., Sofyan, A., Ridwan, R., Hassim, H.A., Respati, A.N., Wardani, W.W., Sadarman, Astuti,
W.D., Jayanegara, A., 2021. Effects of different lactic acid bacteria groups and fibrolytic
Jo

enzymes as additives on silage quality: A meta-analysis. Bioresour. Technol. Reports 14.

https://doi.org/10.1016/j.biteb.2021.100654
Islam, M.T., Alam, M.M., Zoccola, M., 2013. Review on modification of nanocellulose for
application in composites. Int. J. Innov. Res. Sci. Eng. Technol. 2, 5444–5451.
Jafarzadeh, S., Jafari, S.M., Salehabadi, A., Nafchi, A.M., Uthaya Kumar, U.S., Khalil, H.P.S.A.,
2020. Biodegradable green packaging with antimicrobial functions based on the bioactive
compounds from tropical plants and their by-products. Trends Food Sci. Technol.
https://doi.org/10.1016/j.tifs.2020.04.017
Jaiswal, A.K., Kumar, V., Khakalo, A., Lahtinen, P., Solin, K., Pere, J., Toivakka, M., 2021.
Rheological behavior of high consistency enzymatically fibrillated cellulose suspensions.
Cellulose 28, 2087–2104. https://doi.org/10.1007/s10570-021-03688-y
 Journal Pre-proof

Jeoh, T., Cardona, M.J., Karuna, N., Mudinoor, A.R., Nill, J., 2017. Mechanistic kinetic models of
enzymatic cellulose hydrolysis: A review. Biotechnol. Bioeng. 114, 1369–1385.
https://doi.org/10.1002/bit.26277
Jeong, T.S., Choi, C.H., Lee, J.Y., Oh, K.K., 2012. Behaviors of glucose decomposition during acid-
catalyzed hydrothermal hydrolysis of pretreated Gelidium amansii. Bioresour. Technol. 116.
https://doi.org/10.1016/j.biortech.2012.03.104
Johnston, E.T., Lim, P.E., Buhari, N., Keil, E.J., Djawad, M.I., Vis, M.L., 2014. Diversity of
freshwater red algae (Rhodophyta) in Malaysia and Indonesia from morphological and
molecular data. Phycologia 53. https://doi.org/10.2216/13-223.1
Jozala, A.F., de Lencastre-Novaes, L.C., Lopes, A.M., de Carvalho Santos-Ebinuma, V., Mazzola,

of
P.G., Pessoa-Jr, A., Grotto, D., Gerenutti, M., Chaud, M.V., 2016. Bacterial nanocellulose

ro
production and application: a 10-year overview. Appl. Microbiol. Biotechnol. 100, 2063–2072.
https://doi.org/10.1007/s00253-015-7243-4 -p
Kanamarlapudi, S.L.R.K., Chintalpudi, V.K., Muddada, S., 2018. Application of biosorption for
removal of heavy metals from wastewater. Biosorption.
re
https://doi.org/10.5772/intechopen.77315
Kargarzadeh, H., Ahmad, I., Abdullah, I., Dufresne, A., Zainudin, S.Y., Sheltami, R.M., 2012.
lP

Effects of hydrolysis conditions on the morphology, crystallinity, and thermal stability of

cellulose nanocrystals extracted from kenaf bast fibers. Cellulose 19, 855–866.
na

https://doi.org/10.1007/s10570-012-9684-6
Karimi, K., Shafiei, M., Kumar, R., 2013. Progress in physical and chemical pretreatment of
ur

lignocellulosic biomass, in: Gupta, V., Tuohy, M. (Eds.), Biofuel Technologies. Springer Berlin
Heidelberg, Berlin, Heidelberg, pp. 53–96. https://doi.org/10.1007/978-3-642-34519-7_3
Jo

Kavitha, N., Karunya, T.P., Kanchana, S., Mohan, K., Sivaramakrishnan, R., Uthra, S., Kapilan, K.,
Yuvaraj, D., Arumugam, M., 2019. Formulation of alginate based hydrogel from brown
seaweed, Turbinaria conoides for biomedical applications. Heliyon 5, e02916.
https://doi.org/10.1016/j.heliyon.2019.e02916
Khammee, P., Ramaraj, R., Whangchai, N., Bhuyar, P., Unpaprom, Y., 2021. The immobilization of
yeast for fermentation of macroalgae Rhizoclonium sp. for efficient conversion into bioethanol.
Biomass Convers. Biorefinery 11, 827–835. https://doi.org/10.1007/s13399-020-00786-y
Kian, L.K., Saba, N., Jawaid, M., Fouad, H., 2020. Characterization of microcrystalline cellulose
extracted from olive fiber. Int. J. Biol. Macromol. 156.
https://doi.org/10.1016/j.ijbiomac.2020.04.015
Kim, N.J., Li, H., Jung, K., Chang, H.N., Lee, P.C., 2011. Ethanol production from marine algal
 Journal Pre-proof

hydrolysates using Escherichia coli KO11. Bioresour. Technol. 102.

https://doi.org/10.1016/j.biortech.2011.04.071
Klemm, D., Kramer, F., Moritz, S., Lindström, T., Ankerfors, M., Gray, D., Dorris, A., 2011.
Nanocelluloses: A new family of nature-based materials. Angew. Chem. Int. Ed. Engl. 50,
5438–66. https://doi.org/10.1002/anie.201001273
Korzen, L., Pulidindi, I.N., Israel, A., Abelson, A., Gedanken, A., 2015. Single step production of
bioethanol from the seaweed Ulva rigida using sonication. RSC Adv. 5.
https://doi.org/10.1039/c4ra14880k
Kostas, E.T., White, D.A., Cook, D.J., 2020. Bioethanol production from UK seaweeds:
investigating variable pre-treatment and enzyme hydrolysis parameters. Bioenergy Res. 13,

of
271–285. https://doi.org/10.1007/s12155-019-10054-1
Kovacs, T., Naish, V., O’Connor, B., Blaise, C., Gagné, F., Hall, L., Trudeau, V., Martel, P., 2010.

ro
An ecotoxicological characterization of nanocrystalline cellulose (NCC). Nanotoxicology 4.
-p
https://doi.org/10.3109/17435391003628713
Kröger, M., Klemm, M., Nelles, M., 2018. Hydrothermal disintegration and extraction of different
re
microalgae species. Energies 11. https://doi.org/10.3390/en11020450
Kumar, R., Rai, B., Gahlyan, S., Kumar, G., 2021. A comprehensive review on production, surface
lP

modification and characterization of nanocellulose derived from biomass and its commercial
applications. Express Polym. Lett. 15, 104–120.
na

https://doi.org/10.3144/expresspolymlett.2021.11
Kusmono, Listyanda, R.F., Wildan, M.W., Ilman, M.N., 2020. Preparation and characterization of
ur

cellulose nanocrystal extracted from ramie fibers by sulfuric acid hydrolysis. Heliyon 6.
https://doi.org/10.1016/j.heliyon.2020.e05486
Jo

Lee, K.Y., Tammelin, T., Kiiskinen, H., Samela, J., Schlufter, K., Bismarck, A., 2012. Nano-
fibrillated cellulose vs bacterial cellulose: Reinforcing ability of nanocellulose obtained
topdown or bottom-up, in: ECCM 2012 - Composites at Venice, Proceedings of the 15th
European Conference on Composite Materials. Universita di Padova, Venice, Italy, pp. 1–8.
Li, A., Xu, D., Luo, L., Zhou, Yalan, Yan, W., Leng, X., Dai, D., Zhou, Yonghui, Ahmad, H., Rao,
J., Fan, M., 2021. Overview of nanocellulose as additives in paper processing and paper
products. Nanotechnol. Rev. https://doi.org/10.1515/ntrev-2021-0023
Liu, J., Willför, S., Mihranyan, A., 2017a. On importance of impurities, potential leachables and
extractables in algal nanocellulose for biomedical use. Carbohydr. Polym. 172.
https://doi.org/10.1016/j.carbpol.2017.05.002
Liu, Z., Li, X., Xie, W., Deng, H., 2017b. Extraction, isolation and characterization of
 Journal Pre-proof

nanocrystalline cellulose from industrial kelp (Laminaria japonica) waste. Carbohydr. Polym.
173, 353–359. https://doi.org/10.1016/j.carbpol.2017.05.079
Lynd, L.R., Weimer, P.J., van Zyl, W.H., Pretorius, I.S., 2002. Microbial cellulose utilization:
fundamentals and biotechnology. Microbiol. Mol. Biol. Rev. 66.
https://doi.org/10.1128/mmbr.66.4.739.2002
Ma, H., Wang, S., Meng, F., Xu, X., Huo, X., 2017. A hydrazone-carboxyl ligand-linked cellulose
nanocrystal aerogel with high elasticity and fast oil/water separation. Cellulose 24.
https://doi.org/10.1007/s10570-016-1132-6
Mariño, M., Lopes da Silva, L., Durán, N., Tasic, L., 2015. Enhanced materials from nature:
nanocellulose from citrus waste. Molecules 20, 5908–5923.

of
https://doi.org/10.3390/molecules20045908

ro
MarketsandMarkets.com, 2020. Nanocellulose market by type application and region - global
forecast to 2025. https://www.marketsandmarkets.com/Market-Reports/nano-cellulose-market-
-p
56392090.html (accessed 7.30.21).
Martelli-Tosi, M., Torricillas, M.D.S., Martins, M.A., Assis, O.B.G. De, Tapia-Blácido, D.R., 2016.
re
Using commercial enzymes to produce cellulose nanofibers from soybean straw. J. Nanomater.
2016. https://doi.org/10.1155/2016/8106814
lP

Mautner, A., 2020. Nanocellulose water treatment membranes and filters: a review. Polym. Int.
https://doi.org/10.1002/pi.5993
na

Mehanny, S., Abu-El Magd, E.E., Ibrahim, M., Farag, M., Gil-San-Millan, R., Navarro, J., El
Habbak, A.E.H., El-Kashif, E., 2021. Extraction and characterization of nanocellulose from
ur

three types of palm residues. J. Mater. Res. Technol. 10.

https://doi.org/10.1016/j.jmrt.2020.12.027
Jo

Melo, B.C., Paulino, F.A.A., Cardoso, V.A., Pereira, A.G.B., Fajardo, A.R., Rodrigues, F.H.A.,
2018. Cellulose nanowhiskers improve the methylene blue adsorption capacity of chitosan-g-
poly(acrylic acid) hydrogel. Carbohydr. Polym. 181.
https://doi.org/10.1016/j.carbpol.2017.10.079
Metreveli, G., Wågberg, L., Emmoth, E., Belák, S., Strømme, M., Mihranyan, A., 2014. A size-
exclusion nanocellulose filter paper for virus removal. Adv. Healthc. Mater. 3, 1546–1550.
https://doi.org/10.1002/adhm.201300641
Michelin, M., Gomes, D.G., Romaní, A., Polizeli, M. de L.T.M., Teixeira, J.A., 2020. Nanocellulose
production: Exploring the enzymatic route and residues of pulp and paper industry. Molecules
25, 1–36. https://doi.org/10.3390/molecules25153411
Mihranyan, A., 2011. Cellulose from cladophorales green algae: From environmental problem to
 Journal Pre-proof

high-tech composite materials. J. Appl. Polym. Sci. 119, 2449–2460.

https://doi.org/10.1002/app.32959
Mo, L., Pang, H., Tan, Y., Zhang, S., Li, J., 2019. 3D multi-wall perforated nanocellulose-based
polyethylenimine aerogels for ultrahigh efficient and reversible removal of Cu(II) ions from
water. Chem. Eng. J. 378, 122157. https://doi.org/10.1016/j.cej.2019.122157
Moghaddas Kia, E., Ghasempour, Z., Alizadeh, M., 2018. Fabrication of an eco-friendly antioxidant
biocomposite: Zedo gum/sodium caseinate film by incorporating microalgae (Spirulina
platensis). J. Appl. Polym. Sci. 135. https://doi.org/10.1002/app.46024
Mohamed, M.A., Abd Mutalib, M., Mohd Hir, Z.A., M. Zain, M.F., Mohamad, A.B., Jeffery
Minggu, L., Awang, N.A., W. Salleh, W.N., 2017a. An overview on cellulose-based material in

of
tailoring bio-hybrid nanostructured photocatalysts for water treatment and renewable energy

ro
applications. Int. J. Biol. Macromol. https://doi.org/10.1016/j.ijbiomac.2017.05.181
Mohamed, M.A., Salleh, W.N.W., Jaafar, J., Ismail, A.F., Abd Mutalib, M., Mohamad, A.B., Zain,
-p
M.F., Awang, N.A., Mohd Hir, Z.A., 2017b. Physicochemical characterization of cellulose
nanocrystal and nanoporous self-assembled CNC membrane derived from Ceiba pentandra.
re
Carbohydr. Polym. 157. https://doi.org/10.1016/j.carbpol.2016.11.078
Mohammed, N., Grishkewich, N., Berry, R.M., Tam, K.C., 2015. Cellulose nanocrystal–alginate
lP

hydrogel beads as novel adsorbents for organic dyes in aqueous solutions. Cellulose 22, 3725–
3738. https://doi.org/10.1007/s10570-015-0747-3
na

Mustapha, M.U., Halimoon, N., 2015. Microorganisms and biosorption of heavy metals in the
environment: A review paper. J. Microb. Biochem. Technol. 07. https://doi.org/10.4172/1948-
ur

5948.1000219
Naderi, A., Koschella, A., Heinze, T., Shih, K.C., Nieh, M.P., Pfeifer, A., Chang, C.C., Erlandsson,
Jo

J., 2017. Sulfoethylated nanofibrillated cellulose: Production and properties. Carbohydr. Polym.
169. https://doi.org/10.1016/j.carbpol.2017.04.026
Nechyporchuk, O., Belgacem, M.N., Bras, J., 2016. Production of cellulose nanofibrils: A review of
recent advances. Ind. Crops Prod. 93, 2–25. https://doi.org/10.1016/j.indcrop.2016.02.016
Ng, H.M., Sin, L.T., Tee, T.T., Bee, S.T., Hui, D., Low, C.Y., Rahmat, A.R., 2015. Extraction of
cellulose nanocrystals from plant sources for application as reinforcing agent in polymers.
Compos. Part B Eng. 75, 176–200. https://doi.org/10.1016/j.compositesb.2015.01.008
Noguchi, Y., Homma, I., Matsubara, Y., 2017. Complete nanofibrillation of cellulose prepared by
phosphorylation. Cellulose 24. https://doi.org/10.1007/s10570-017-1191-3
Norrrahim, M.N.F., Kasim, N.A.M., Knight, V.F., Misenan, M.S.M., Janudin, N., Shah, N.A.A.,
Kasim, N., Yusoff, W.Y.W., Noor, S.A.M., Jamal, S.H., Ong, K.K., Yunus, W.M.Z.W., 2021.
 Journal Pre-proof

Nanocellulose: a bioadsorbent for chemical contaminant remediation. RSC Adv. 11, 7347–
7368. https://doi.org/10.1039/D0RA08005E
Olivera, S., Muralidhara, H.B., Venkatesh, K., Guna, V.K., Gopalakrishna, K., Kumar K., Y., 2016.
Potential applications of cellulose and chitosan nanoparticles/composites in wastewater
treatment: A review. Carbohydr. Polym. 153, 600–618.
https://doi.org/10.1016/j.carbpol.2016.08.017
Palomer, X., Domínguez-Puigjaner, E., Vendrell, M., Llop-Tous, I., 2004. Study of the strawberry
Cel1 endo-β-(1,4)-glucanase protein accumulation and characterization of its in vitro activity by
heterologous expression in Pichia pastoris. Plant Sci. 167.
https://doi.org/10.1016/j.plantsci.2004.04.017

of
Paniz, O.G., Pereira, C.M.P., Pacheco, B.S., Wolke, S.I., Maron, G.K., Mansilla, A., Colepicolo, P.,

ro
Orlandi, M.O., Osorio, A.G., Carreño, N.L.V., 2020. Cellulosic material obtained from
Antarctic algae biomass. Cellulose 27, 113–126. https://doi.org/10.1007/s10570-019-02794-2
-p
Phanthong, P., Reubroycharoen, P., Hao, X., Xu, G., Abudula, A., Guan, G., 2018. Nanocellulose:
extraction and application. Carbon Resour. Convers. 1, 32–43.
re
https://doi.org/10.1016/j.crcon.2018.05.004
Pitkänen, M., Kangas, H., Laitinen, O., Sneck, A., Lahtinen, P., Peresin, M.S., Niinimäki, J., 2014.
lP

Characteristics and safety of nano-sized cellulose fibrils. Cellulose 21.

https://doi.org/10.1007/s10570-014-0397-x
na

Polizeli, M.L.T.M., Rizzatti, A.C.S., Monti, R., Terenzi, H.F., Jorge, J.A., Amorim, D.S., 2005.
Xylanases from fungi: Properties and industrial applications. Appl. Microbiol. Biotechnol.
ur

https://doi.org/10.1007/s00253-005-1904-7
Qiao, H., Zhou, Y., Yu, F., Wang, E., Min, Y., Huang, Q., Pang, L., Ma, T., 2015. Effective removal
Jo

of cationic dyes using carboxylate-functionalized cellulose nanocrystals. Chemosphere 141.

https://doi.org/10.1016/j.chemosphere.2015.07.078
Qing, Y., Sabo, R., Zhu, J.Y., Agarwal, U., Cai, Z., Wu, Y., 2013. A comparative study of cellulose
nanofibrils disintegrated via multiple processing approaches. Carbohydr. Polym. 97.
https://doi.org/10.1016/j.carbpol.2013.04.086
Reshmy, R., Thomas, D., Philip, E., Paul, S.A., Madhavan, A., Sindhu, R., Binod, P., Pugazhendhi,
A., Sirohi, R., Tarafdar, A., Pandey, A., 2021. Potential of nanocellulose for wastewater
treatment. Chemosphere 281, 130738. https://doi.org/10.1016/j.chemosphere.2021.130738
Rajesh Kumar Dora, T., Ghosh, S., Damodar, R., 2020. Synthesis and evaluation of physical
properties of agar biopolymer film coating-an alternative for food packaging industry. Mater.
Res. Express 7. https://doi.org/10.1088/2053-1591/abb7ac
 Journal Pre-proof

Rasalingam, S., Peng, R., Koodali, R.T., 2014. Removal of hazardous pollutants from wastewaters:
Applications of TiO2-SiO2 mixed oxide materials. J. Nanomater.
https://doi.org/10.1155/2014/617405
Rashid, E.S.A., Gul, A., Yehya, W.A.H., Julkapli, N.M., 2021. Physico-chemical characteristics of
nanocellulose at the variation of catalytic hydrolysis process. Heliyon 7, e07267.
https://doi.org/10.1016/j.heliyon.2021.e07267
Resch, A., Staud, C., Radtke, C., 2021. Nanocellulose-based wound dressing for conservative wound
management in children with second-degree burns. Int. Wound J.
https://doi.org/10.1111/iwj.13548
Reshmy, R., Madhavan, A., Philip, E., Paul, S.A., Sindhu, R., Binod, P., Pugazhendhi, A., Sirohi, R.,

of
Pandey, A., 2021. Sugarcane bagasse derived nanocellulose reinforced with frankincense

ro
(Boswellia serrata): Physicochemical properties, biodegradability and antimicrobial effect for
controlling microbial growth for food packaging application. Environ. Technol. Innov. 21.
-p
https://doi.org/10.1016/j.eti.2020.101335
Rhim, J.W., 2011. Effect of clay contents on mechanical and water vapor barrier properties of agar-
re
based nanocomposite films. Carbohydr. Polym. 86.
https://doi.org/10.1016/j.carbpol.2011.05.010
lP

Ribeiro, R.S.A., Pohlmann, B.C., Calado, V., Bojorge, N., Pereira, N., 2019. Production of
nanocellulose by enzymatic hydrolysis: Trends and challenges. Eng. Life Sci. 19, 279–291.
na

https://doi.org/10.1002/elsc.201800158
Rocha, F.D., Soares, A.R., Houghton, P.J., Pereira, R.C., Kaplan, M.A.C., Teixeira, V.L., 2007.
ur

Potential cytotoxic activity of some Brazilian seaweeds on human melanoma cells. Phyther.
Res. 21. https://doi.org/10.1002/ptr.2038
Jo

Rodrigues, D., Freitas, A.C., Pereira, L., Rocha-Santos, T.A.P., Vasconcelos, M.W., Roriz, M.,
Rodríguez-Alcalá, L.M., Gomes, A.M.P., Duarte, A.C., 2015. Chemical composition of red,
brown and green macroalgae from Buarcos bay in Central West Coast of Portugal. Food Chem.
183, 197–207. https://doi.org/10.1016/j.foodchem.2015.03.057
Sabaruddin, F.A., Paridah, M.T., Lee Seng Hua, Lee Chuan Li, 2021. The sulphate removal via post
alkaline treatment on nanocrystalline cellulose with different lignin content extracted from
kenaf core. J. Adv. Res. Fluid Mech. Therm. Sci. 84, 11–19.
https://doi.org/10.37934/arfmts.84.1.1119
Sabaruddin, F.A., Paridah, M.T., Sapuan, S.M., Ilyas, R.A., Lee, S.H., Abdan, K., Mazlan, N.,
Roseley, A.S.M., Abdul Khalil, H.P.S., 2020. The effects of unbleached and bleached
nanocellulose on the thermal and flammability of polypropylene-reinforced kenaf core hybrid
 Journal Pre-proof

polymer bionanocomposites. Polymers (Basel). 13, 116.

https://doi.org/10.3390/polym13010116
Satyamurthy, P., Vigneshwaran, N., 2013. A novel process for synthesis of spherical nanocellulose
by controlled hydrolysis of microcrystalline cellulose using anaerobic microbial consortium.
Enzyme Microb. Technol. 52. https://doi.org/10.1016/j.enzmictec.2012.09.002
Seghetta, M., Romeo, D., D’Este, M., Alvarado-Morales, M., Angelidaki, I., Bastianoni, S.,
Thomsen, M., 2017. Seaweed as innovative feedstock for energy and feed – Evaluating the
impacts through a Life Cycle Assessment. J. Clean. Prod. 150.
https://doi.org/10.1016/j.jclepro.2017.02.022
Shak, K.P.Y., Pang, Y.L., Mah, S.K., 2018. Nanocellulose: Recent advances and its prospects in

of
environmental remediation. Beilstein J. Nanotechnol. 9, 2479–2498.

ro
https://doi.org/10.3762/bjnano.9.232
Shang, Q., Chen, J., Hu, Y., Yang, X., Hu, L., Liu, C., Ren, X., Zhou, Y., 2021. Facile fabrication of
-p
superhydrophobic cross-linked nanocellulose aerogels for oil–water separation. Polymers
(Basel). 13. https://doi.org/10.3390/polym13040625
re
Sharma, M., Aguado, R., Murtinho, D., Valente, A.J.M., Mendes De Sousa, A.P., Ferreira, P.J.T.,
2020. A review on cationic starch and nanocellulose as paper coating components. Int. J. Biol.
lP

Macromol. https://doi.org/10.1016/j.ijbiomac.2020.06.131
Shatkin, J.A., Kim, B., 2015. Cellulose nanomaterials: life cycle risk assessment, and environmental
na

health and safety roadmap. Environ. Sci. Nano 2, 477–499. https://doi.org/10.1039/c5en00059a

Shen, R., Xue, S., Xu, Y., Liu, Q., Feng, Z., Ren, H., Zhai, H., Kong, F., 2020. Research progress
ur

and development demand of nanocellulose reinforced polymer composites. Polymers (Basel).

12, 2113. https://doi.org/10.3390/polym12092113
Jo

Shree, P., Mishra, P., Selvaraj, C., Singh, S.K., Chaube, R., Garg, N., Tripathi, Y.B., 2020. Targeting
COVID-19 (SARS-CoV-2) main protease through active phytochemicals of ayurvedic
medicinal plants–Withania somnifera (Ashwagandha), Tinospora cordifolia (Giloy) and
Ocimum sanctum (Tulsi)–a molecular docking study. J. Biomol. Struct. Dyn.
https://doi.org/10.1080/07391102.2020.1810778
Singh, A., Bajar, S., Devi, A., Pant, D., 2021. An overview on the recent developments in fungal
cellulase production and their industrial applications. Bioresour. Technol. Reports.
https://doi.org/10.1016/j.biteb.2021.100652
Singh, S., Gaikwad, K.K., Park, S. Il, Lee, Y.S., 2017. Microwave-assisted step reduced extraction of
seaweed (Gelidiella aceroso) cellulose nanocrystals. Int. J. Biol. Macromol. 99, 506–510.
https://doi.org/10.1016/j.ijbiomac.2017.03.004
 Journal Pre-proof

Squinca, P., Bilatto, S., Badino, A.C., Farinas, C.S., 2020. Nanocellulose production in future
biorefineries: an integrated approach using tailor-made enzymes. ACS Sustain. Chem. Eng. 8,
2277–2286. https://doi.org/10.1021/acssuschemeng.9b06790
Starko, S., Mansfield, S.D., Martone, P.T., 2018. Cell wall chemistry and tissue structure underlie
shifts in material properties of a perennial kelp. Eur. J. Phycol. 53.
https://doi.org/10.1080/09670262.2018.1449013
Stoudmann, N., Schmutz, M., Hirsch, C., Nowack, B., Som, C., 2020. Human hazard potential of
nanocellulose: quantitative insights from the literature. Nanotoxicology 14, 1241–1257.
https://doi.org/10.1080/17435390.2020.1814440
Sudhakar, M.P., Kumar, B.R., Mathimani, T., Arunkumar, K., 2019. A review on bioenergy and

of
bioactive compounds from microalgae and macroalgae-sustainable energy perspective. J. Clean.

ro
Prod. https://doi.org/10.1016/j.jclepro.2019.04.287
Sun, H., Wang, W., Li, W., Zhang, X., 2014. Preparation technique of Eichhornia crassipes-based
-p
composite packaging materials. Nongye Gongcheng Xuebao/Transactions Chinese Soc. Agric.
Eng. 30. https://doi.org/10.3969/j.issn.1002-6819.2014.12.032
re
Taheri, H., Samyn, P., 2016. Effect of homogenization (microfluidization) process parameters in
mechanical production of micro- and nanofibrillated cellulose on its rheological and
lP

morphological properties. Cellulose 23. https://doi.org/10.1007/s10570-016-0866-5

Tan, H.F., Ooi, B.S., Leo, C.P., 2020. Future perspectives of nanocellulose-based membrane for
na

water treatment. J. Water Process Eng. https://doi.org/10.1016/j.jwpe.2020.101502

Thomas, B., Raj, M.C., Athira, B.K., Rubiyah, H.M., Joy, J., Moores, A., Drisko, G.L., Sanchez, C.,
ur

2018. Nanocellulose, a versatile green platform: from biosources to materials and their
applications. Chem. Rev. https://doi.org/10.1021/acs.chemrev.7b00627
Jo

Thomas, P., Duolikun, T., Rumjit, N.P., Moosavi, S., Lai, C.W., Bin Johan, M.R., Fen, L.B., 2020.
Comprehensive review on nanocellulose: Recent developments, challenges and future
prospects. J. Mech. Behav. Biomed. Mater. 110, 103884.
https://doi.org/10.1016/j.jmbbm.2020.103884
Thongekkaew, J., Kongsanthia, J., 2016. Screening and identification of cellulase producing yeast
from Rongkho forest, Ubon Ratchathani University. Bioeng. Biosci. 4.
https://doi.org/10.13189/bb.2016.040301
Tokay, B., Akpınar, I., 2021. A comparative study of heavy metals removal using agricultural waste
biosorbents. Bioresour. Technol. Reports 15, 100719.
https://doi.org/10.1016/j.biteb.2021.100719
Touijer, H., Benchemsi, N., Ettayebi, M., Janati Idrissi, A., Chaouni, B., Bekkari, H., 2019.
 Journal Pre-proof

Thermostable cellulases from the yeast Trichosporon sp. Enzyme Res. 2019.
https://doi.org/10.1155/2019/2790414
Trache, D., Tarchoun, A.F., Derradji, M., Hamidon, T.S., Masruchin, N., Brosse, N., Hussin, M.H.,
2020. Nanocellulose: From fundamentals to advanced applications. Front. Chem. 8, 1–33.
https://doi.org/10.3389/fchem.2020.00392
Trache, D., Thakur, V.K., 2020. Nanocellulose and nanocarbons based hybrid materials: Synthesis,
characterization and applications. Nanomaterials 10, 1–5.
https://doi.org/10.3390/nano10091800
Tsukamoto, J., Durán, N., Tasic, L., 2013. Nanocellulose and bioethanol production from orange
waste using isolated microorganisms. J. Braz. Chem. Soc. 24, 1537–1543.

of
https://doi.org/10.5935/0103-5053.20130195

ro
Uetani, K., Yano, H., 2011. Nanofibrillation of wood pulp using a high-speed blender.
Biomacromolecules 12. https://doi.org/10.1021/bm101103p
-p
Van Hai, L., Son, H.N., Seo, Y.B., 2015. Physical and bio-composite properties of nanocrystalline
cellulose from wood, cotton linters, cattail, and red algae. Cellulose 22, 1789–1798.
re
https://doi.org/10.1007/s10570-015-0633-z
Veluchamy, C., Palaniswamy, R., 2020. A review on marine algae and its applications. Asian J.
lP

Pharm. Clin. Res. 13, 21–27. https://doi.org/10.22159/ajpcr.2020.v13i3.36130

Venkatesan, J., Anil, S., Kim, S.K., Shim, M.S., 2016. Seaweed polysaccharide-based nanoparticles:
na

Preparation and applications for drug delivery. Polymers (Basel).

https://doi.org/10.3390/polym8020030
ur

Ventura-Cruz, S., Tecante, A., 2021. Nanocellulose and microcrystalline cellulose from agricultural
waste: Review on isolation and application as reinforcement in polymeric matrices. Food
Jo

Hydrocoll. 118, 106771. https://doi.org/10.1016/j.foodhyd.2021.106771

Ventura, C., Lourenço, A.F., Sousa-Uva, A., Ferreira, P.J.T., Silva, M.J., 2018. Evaluating the
genotoxicity of cellulose nanofibrils in a co-culture of human lung epithelial cells and
monocyte-derived macrophages. Toxicol. Lett. 291. https://doi.org/10.1016/j.toxlet.2018.04.013
Vigneshwaran, N., Satyamurthy, P., 2016. Nanocellulose production using cellulose degrading fungi,
in: Prasad, R. (Ed.), Advances and Applications Through Fungal Nanobiotechnology. Springer
International Publishing Switzerland, Switzerland, pp. 321–331. https://doi.org/10.1007/978-3-
319-42990-8_16
Voisin, H., Bergström, L., Liu, P., Mathew, A.P., 2017. Nanocellulose-based materials for water
purification. Nanomaterials 7. https://doi.org/10.3390/nano7030057
Wågberg, L., Decher, G., Norgren, M., Lindström, T., Ankerfors, M., Axnäs, K., 2008. The build-up
 Journal Pre-proof

of polyelectrolyte multilayers of microfibrillated cellulose and cationic polyelectrolytes.

Langmuir 24. https://doi.org/10.1021/la702481v
Wahlström, N., Edlund, U., Pavia, H., Toth, G., Jaworski, A., Pell, A.J., Choong, F.X., Shirani, H.,
Nilsson, K.P.R., Richter-Dahlfors, A., 2020. Cellulose from the green macroalgae Ulva lactuca:
isolation, characterization, optotracing, and production of cellulose nanofibrils. Cellulose 27.
https://doi.org/10.1007/s10570-020-03029-5
Wang, Q.Q., Zhu, J.Y., Gleisner, R., Kuster, T.A., Baxa, U., McNeil, S.E., 2012. Morphological
development of cellulose fibrils of a bleached eucalyptus pulp by mechanical fibrillation.
Cellulose 19. https://doi.org/10.1007/s10570-012-9745-x
Wang, R., Guan, S., Sato, A., Wang, X., Wang, Z., Yang, R., Hsiao, B.S., Chu, B., 2013.

of
Nanofibrous microfiltration membranes capable of removing bacteria, viruses and heavy metal

ro
ions. J. Memb. Sci. 446, 376–382. https://doi.org/10.1016/j.memsci.2013.06.020
Wang, X., Yeh, T.-M., Wang, Z., Yang, R., Wang, R., Ma, H., Hsiao, B.S., Chu, B., 2014.
-p
Nanofiltration membranes prepared by interfacial polymerization on thin-film nanofibrous
composite scaffold. Polymer (Guildf). 55, 1358–1366.
re
https://doi.org/10.1016/j.polymer.2013.12.007
Wijesinghe, W.A.J.P., Jeon, Y.J., 2012. Enzyme-assistant extraction (EAE) of bioactive components:
lP

A useful approach for recovery of industrially important metabolites from seaweeds: A review.
Fitoterapia. https://doi.org/10.1016/j.fitote.2011.10.016
na

Xiang, Z., Gao, W., Chen, L., Lan, W., Zhu, J.Y., Runge, T., 2016. A comparison of cellulose
nanofibrils produced from Cladophora glomerata algae and bleached eucalyptus pulp.
ur

Cellulose 23, 493–503. https://doi.org/10.1007/s10570-015-0840-7

Xing, J., Tao, P., Wu, Z., Xing, C., Liao, X., Nie, S., 2019. Nanocellulose-graphene composites: A
Jo

promising nanomaterial for flexible supercapacitors. Carbohydr. Polym.

https://doi.org/10.1016/j.carbpol.2018.12.010
Xu, S.Y., Huang, X., Cheong, K.L., 2017. Recent advances in marine algae polysaccharides:
Isolation, structure, and activities. Mar. Drugs 15, 1–16. https://doi.org/10.3390/md15120388
Yassin, M.A., Gad, A.A.M., Ghanem, A.F., Abdel Rehim, M.H., 2019. Green synthesis of cellulose
nanofibers using immobilized cellulase. Carbohydr. Polym. 205.
https://doi.org/10.1016/j.carbpol.2018.10.040
Yazdani, P., Karimi, K., Taherzadeh, M.J., 2011. Improvement of enzymatic hydrolysis of a marine
macro-alga by dilute acid hydrolysis pretreatment. Proc. World Renew. Energy Congr. –
Sweden, 8–13 May, 2011, Linköping, Sweden 57, 186–191.
https://doi.org/10.3384/ecp11057186
 Journal Pre-proof

Yoon, J.J., Kim, Y.J., Kim, S.H., Ryu, H.J., Choi, J.Y., Kim, G.S., Shin, M.K., 2010. Production of
polysaccharides and corresponding sugars from red seaweed, in: Advanced Materials Research.
pp. 463–466. https://doi.org/10.4028/www.scientific.net/AMR.93-94.463
Yu, S., Sun, J., Shi, Y., Wang, Q., Wu, J., Liu, J., 2021. Nanocellulose from various biomass wastes:
Its preparation and potential usages towards the high value-added products. Environ. Sci.
Ecotechnology 5, 100077. https://doi.org/10.1016/j.ese.2020.100077
Yuan, M., Xiao, H., Wang, R., Duan, Y., Cao, Q., 2019. Effects of changes in precipitation pattern
and of seaweed fertilizer addition on plant traits and biological soil crusts. J. Appl. Phycol. 31.
https://doi.org/10.1007/s10811-019-01838-1
Zanchetta, E., Damergi, E., Patel, B., Borgmeyer, T., Pick, H., Pulgarin, A., Ludwig, C., 2021. Algal

of
cellulose, production and potential use in plastics: Challenges and opportunities. Algal Res. 56,

ro
102288. https://doi.org/10.1016/j.algal.2021.102288
Zhang, W., Zhang, Y., Cao, J., Jiang, W., 2021. Improving the performance of edible food packaging
-p
films by using nanocellulose as an additive. Int. J. Biol. Macromol.
https://doi.org/10.1016/j.ijbiomac.2020.10.185
re
Zhou, W., Fang, J., Tang, S., Wu, Z., Wang, X., 2021. 3D-Printed nanocellulose-based cushioning–
antibacterial dual-function food packaging aerogel. Molecules 26.
lP

https://doi.org/10.3390/molecules26123543
Zinge, C., Kandasubramanian, B., 2020. Nanocellulose based biodegradable polymers. Eur. Polym.
na

J. 133, 109758. https://doi.org/10.1016/j.eurpolymj.2020.109758

Zulfiana, D., Karimah, A., Anita, S.H., Masruchin, N., Wijaya, K., Suryanegara, L., Fatriasari, W.,
ur

Fudholi, A., 2020. Antimicrobial Imperata cylindrica paper coated with anionic nanocellulose
crosslinked with cationic ions. Int. J. Biol. Macromol. 164.
Jo

https://doi.org/10.1016/j.ijbiomac.2020.07.102
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Declaration of interests

☒ The authors declare that they have no known competing financial interests or personal relationships that
could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered as
potential competing interests:

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Graphical abstract
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Highlights
 Marine algae have high concentration of carbohydrate can utilize as nanocellulose
 Microbial treatment for nanocellulose extraction can reduce the usage of chemicals
 Nanocellulose extraction through microbial treatment used enzyme known as cellulase
 Cellulase produced through various microorganisms such as bacteria, yeast and fungi
 Nanocellulose derived marine algae have been utilized in wide range of applications

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Graphics Abstract
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