1.

Introduction
Fossil resources have changed the world's energy supply, but their depletion and global
warming have raised concerns about their sustainability. To address this issue, people are
looking for renewable alternatives to replace fossil fuels. Throughout years; solar, wind,
biomass, hydroelectric and geothermal energy systems have been developed. Biomass is the
only sustainable source of organic carbon on the planet, and is seen as a solution for
producing fuels and chemicals [1].
Biofuels are fuels obtained from biomass that can be restored fast. Biomass could be found
throughout the earth, and it usually takes the form of organic materials like grass, wood,
agricultural products, and their leftovers and garbage. These biofuels are classified into three
generation; first generation biofuels come from food crops and are made through
fermentation [2]. Second-generation biofuels are manufactured from lignocellulosic biomass
sources such as agricultural residue or energy crops cultivated on unproductive land [3].
There is also third-generation biofuel which comes from algae and other microorganisms
with potential for carbon sequestration and can use non-arable land and waste resources [4].
Second generation biofuels are widely regarded as more efficient, cost-effective, and
environmentally sustainable compared to both first and third generation biofuels. This is
primarily due to their ability to avoid competition with food production by utilizing waste
materials as feedstock. Furthermore, second generation biofuels are less expensive to produce
and can be easily manufactured in large quantities The increasing demand for renewable
sources of energy has led to significant research in the field of cellulose hydrolysis. The
conversion of cellulose into fermentable sugars is crucial for the production of biofuels and
value-added chemicals [3].
Cellulose is a complex polysaccharide essential for plant cell wall structural integrity and is
used for producing second-generation biofuels, particularly cellulosic ethanol, by fermenting
sugars from cellulose and hemicellulose fractions. It is made up of a long glucose molecule
with β-1,4-glycosidic bonds, offering high tensile strength, stiffness, and insolubility in water
and organic solvents. It resists chemical degradation, including digestion by many organisms.
Cellulose's multifunctionality is evident in its applications in different areas of industry
especially in biofuel production [5]. Therefore, Hydrolysis of cellulose is very critical for
biofuel production, because only glucose, not cellulose, can be consumed by the bacteria used
in fermentation to produce biofuel. The increasing demand for renewable sources of energy
has led to significant research in the field of cellulose hydrolysis.
The conversion of cellulose into fermentable sugars is crucial for the production of biofuels
and value-added chemicals [6], [7]. Cellulose hydrolysis refers to the enzymatic or chemical
breakdown of cellulose into its constituent glucose molecules. There are several techniques
available for the hydrolysis of cellulose, including acid hydrolysis, enzymatic hydrolysis, and
solid acid catalysis. Acid hydrolysis involves the use of mineral acids such as HCl and H 2SO4
to break down cellulose into glucose. However, this method has certain drawbacks, such as
difficulties in separating the products, corrosion of the reactor, limited recyclability of the
catalyst, and challenges in treating the waste effluent [5]. On the other hand, cellulose is
catalyzed by cellulase enzymes, but their low efficiency, limited robustness, reduced
recyclability, enzyme deactivation and high cost make them unsuitable for many applications
[8]. Alternatively, solid acid catalysts have shown promise in the hydrolysis of cellulose into

glucose. These catalysts can overcome some of the limitations of traditional acid catalysts.
Solid acids can also be more easily separated from the reaction mixture, allowing for easier
purification of the final product. Some examples of solid acid catalysts used in cellulose
hydrolysis include sulfonated carbon-based materials, zeolites, and acidic ionic liquids.
Metal-organic framework materials, hetero polyacid, sulfonated zirconia, and sulfonated
nano-graphene are examples of existing solid acid catalysts [6]. Among the various catalysts,
metal-organic frameworks (MOFs) have emerged as promising candidates due to their high
stability, porosity, and better catalytic efficiency [9].
Metal-organic frameworks (MOFs) are a series of crystalline porous materials that are
coordinated to organic ligands by metal ions or clusters [9], [10]. MOFs are attractive for
applications in gas storage, separations, catalysis, sensing, drug delivery, and many other
fields due to their extraordinarily high surface areas, tunable pore sizes and shapes, and high
porosity [11]–[15]. The synthesis of MOFs typically involves mixing the metal ion or cluster
with the organic ligand in a suitable solvent under controlled conditions, such as temperature
and pressure. The resulting solution is then heated, cooled, or allowed to sit for a certain
period of time, depending on the specific synthesis protocol used. One of the key factors in
determining the properties and performance of MOFs is the choice of metal and ligand.
Different combinations of metals and ligands can lead to MOFs with varying pore sizes,
surface areas, and chemical reactivities, among other properties [16]. For better efficiency
and great mass transfer metal organic frameworks has to be modified to improve porosity and
surface area. Researchers have developed a variety of methods for surface modification and
porosity enlargement due to the structural designability of MOFs; functionalization by amine
and imine is one example of surface modification techniques of MOFs. There are several
methods for increasing porosity, however stability and porosity are incompatible, making
them difficult to achieve. For the rational design and controlled synthesis of many
hierarchically porous metal organic frameworks (HP-MOFs) with outstanding stability, G.
Cai and H.L. Jiang developed a simple and versatile modulator-induced defect-formation
technique, in which pore sizes and pore volumes may be easily modified [17]. The
modulator plays a dual role: the carboxylic acid coordinates to the metal ion for the formation
of metal–oxo clusters, while the alkyl chain creates structural defects and additional pore
space [18], [19].
Additionally, introducing magnetic nanoparticles into the MOF structure, the resulting
material can exhibit magnetic properties which can be useful for various applications such as
recovery, sensing, energy storage, and drug delivery [20]. The synthesis of magnetically
modified MOFs involves the incorporation of magnetic species such as iron oxide
nanoparticles or magnetic metal ions into the MOF structure during the synthesis process.
The magnetic particles or ions are typically introduced along with the metal ions or organic
ligands that make up the MOF. The resulting material exhibits both the properties of the
MOF and the magnetic species, providing enhanced functionality [21]. Magnetizing Metal-
Organic Frameworks (MMOFs) with magnetic nanoparticles can enhance the efficiency of
cellulose hydrolysis applications by providing a means of separating the MOF catalyst from
the reaction mixture using an external magnetic field. This allows for easier catalyst recovery
and recyclability, resulting in reduced costs and environmental impact [22]. Among many we
select Cu-MOF because it is a superior catalytic performance in cellulose hydrolysis reactions
due to its high surface area and pore volume, high stability under harsh reaction conditions,
and the presence of copper ions in the framework. These features make it a promising
candidate for intended applications. Additionally, the presence of copper ions in the
framework facilitates the activation of cellulose substrates through selective redox reactions,
leading to more efficient hydrolysis of cellulose [23]. Herein, we proposed synthesis of
MOFs with magnetically modified and modulate-induced defect strategy for recovery of
MOFs and high porosity with better stability for hydrolysis of cellulose. This research could
potentially lead to an increase in the efficiency of cellulose hydrolysis, thus reducing the cost
and energy input required for recovery of MOFs from reaction medium and increase mass
transfer into MOFs matrix. Additionally, the development of efficient and recyclable
catalysts could decrease environmental impact by reducing the amount of waste generated
from the process.
 1.1. Statement of the Problem
Conventional energy sources, such as coal, petroleum, and natural gas, are non-renewable
and limited in quantity. They face high production costs, transmission and distribution losses,
limited reach, environmental and public health threats, global warming, greenhouse effects,
acid rain, and extraction time. Alternative energy sources, such as solar, wind, hydro,
geothermal, and biomass are renewable, naturally replenished, and environmentally friendly
compared to conventional energy sources. Biomass is a sustainable energy source made from
organic matter like plants and agricultural waste, reducing greenhouse gas emissions and
environmental impacts. It is also locally produced, reducing dependence on fossil fuel energy
sources [1]–[4].
However, the hydrolysis of complex polysaccharides into simple sugars is a time-consuming
and costly process. The current strategies involve the use of inorganic catalysts under harsh
reaction conditions, which not only affect the yield but also raise environmental concerns or
use enzyme for conversion of polymers like cellulose to fermentable sugars [5]. Therefore,
there is an unmet need for sustainable and highly efficient heterogeneous catalysts with easy
recyclability that can potentially overcome the existing limitations. The conversion of
cellulose into its individual sugar components for the production of biofuels and other high-
value products is an important area of research. One promising approach involves using UiO-
66 MOF to selectively treat biomass to produce bioethanol. The UiO-66 MOF's catalytic
potential was investigated using carboxyl methyl cellulose (CMC) and sucrose as models.
However, the MOF's contribution to polymeric CMC hydrolysis was insignificant at a fixed
temperature, possibly due to its large molecular size [24]. Therefore, the small porosity and
low surface area of the MOFs make them unfavourable for cellulose, because it leads to low
mass transfer of cellulose to MOFs pore. So, to overcome such problems modifying the
catalyst pore, surface area and gaining or modification of MOFs with magnetic particles
would improve their efficiency and recyclability.
 1.2. Significance of the Project
The significance of this research lies in the development of new and efficient catalysts for
cellulose hydrolysis, which is a crucial step in the production of biofuels and chemicals. The
use of magnetically modified Cu-MOFs expected to overcome the limitations of non-
magnetic Cu-MOFs, such as poor stability and difficulty in separation and recycling. The
porosity and surface area of the MOF is determinant factor for mass transfer, hence HP-Cu-
MOF expected to address the issue of mass transfer limitation that prevailed in most parent
MOFs. The findings of this research can have important implications for the development of
sustainable processes for the production of biofuels and chemicals from renewable resources.

1.3. Scope of the study

The scope of this research is developing simple, rapid, and low-cost metal-organic
frameworks-based catalyst for hydrolysis of cellulose to fermentable sugars. The cellulose is
applicable for production of fermentable sugars like glucose and xylanose. In this work, α-
cellulose will be used for conversion of sugars using the developed technique.

1.4. Objective
1.4.1. General Objective
The main objective of this research is to synthesize parent and modified Cu-MOFs for the
catalytic hydrolysis of Cellulose.
1.4.2. Specific Objectives
1. To synthesize parent Cu-MOFs, HP-Cu-MOFs and Fe 3O4 modified hydrothermal
method.
2. To characterize the physical and chemical properties of the synthesized magnetic Cu-
MOFs using various.
3. To evaluate and compare the catalytic performance of the synthesized Cu-MOFs in
the hydrolysis of cellulose.
4. To investigate the reusability and stability of the magnetic Cu-MOF catalysts in
cellulose hydrolysis.
 2. Literature Review
2.1. Introduction
The development of sustainable and renewable sources of energy has become a major global
concern due to the increasing demand for energy and the depletion of fossil fuels. Biomass is
a promising alternative source of energy that can be converted into fuels and chemicals
through a variety of processes such as hydrolysis, pyrolysis, and gasification. However, the
low reactivity and high stability of biomass pose significant challenges for its conversion into
value-added products [2] – [4]. Cellulose hydrolysis is the process of breaking down cellulose
molecules into simple sugars, also known as monosaccharides, using enzymes called
cellulases. This process is important for biofuel production because only glucose, not
cellulose, can be consumed by the bacteria used in fermentation to produce biofuel. Research
shows that cellulase enzymes significantly reduce the costs of plant biomass breakdown,
making hydrolysis of cellulose economically important [6], [7]. Metal-organic frameworks
(MOFs) have emerged as a promising class of materials for catalysing the conversion of
cellulose due to their high surface area, tunable porosity, and tailorable chemical properties.
In particular, copper-based MOFs (Cu-MOFs) have been shown to exhibit excellent catalytic
activity in the conversion of cellulose, hemicellulose, and lignin into value-added chemicals.
However, the low magnetic properties of Cu-MOFs limit their separation and recycling from
reaction mixtures, which hinders their practical applications [9]. To address this issue,
researchers have developed magnetically modified MOFs by incorporating magnetic
nanoparticles into the MOF framework [25]. This literature review will provide an overview
of the recent advances in the synthesis and application of magnetically modified MOFs for
biomass hydrolysis.

2.2. Synthesis of magnetically modified Cu-MOFs

2.2.1. In situ magnetization method
The process involves adding magnetic particles and organic precursors to a reaction solution,
which undergoes a nucleation and growth process to form the magnetic framework. The
magnetic particles are dispersed throughout the framework material, resulting in a
homogenous distribution. Fe3O4 @SiO2–MIL-101 was obtained by mixing MIL-101 and
silica-coated Fe3O4 microparticles in a sample solution under sonication to achieve in situ
magnetization of MIL-101 microcrystals as shown on figure blow. A facile magnetization of
MOF MIL-101 for rapid magnetic solid-phase extraction of polycyclic aromatic
hydrocarbons (PAHs) from environmental water samples. The developed approach offers
large enrichment factor, wide linear range, and good reproducibility. Owing to the diverse
structures and unique characteristics of MOFs, the combination of magnetic separation with
MOFs is attractive for rapid and efficient extraction of various trace analytes from
environmental and biological samples [26].

Figure 1:Schematic diagram for the application of Fe 3O4@SiO2–MIL-101 MNPs as sorbent for MSP
[26]

The advantages of this method are that it can simplify the synthesis process, reduce the use of
solvents, allows for the simultaneous preparation of magnetic MOFs and their application in
magnetic solid-phase extraction and achieve a strong interaction between magnetic
nanoparticles and MOFs. It is challenging to avoid direct nucleation and growth of metal-
organic frameworks on magnetic nanoparticles during in situ growth. This results in some
magnetic nanoparticles getting embedded in the pores of the framework, making it difficult to
achieve composites with desired surface area, uniform distribution of magnetic nanoparticles,
and large pore volume. In addition, poor control over the particle size and morphology, and
possible aggregation of magnetic nanoparticles [26]

2.2.2. In situ -One-step synthesis

In situ - One-step hydrothermal synthesis of magnetic MOF refers to a chemical process that
involves synthesizing magnetic metal-organic frameworks (MMOFs) under hydrothermal
conditions. This method involves using a one-step hydrothermal strategy, which means that
the material is synthesized in a single step using high temperature water. The magnetic
properties of the synthesized MOF make it useful for various applications as it is easily
recovered from reaction mixture. This method is advantageous because it combines the
synthesis and modification of MOFs in one step, leading to a simplified process with
improved efficiency.
Aslam et al. first time developed this new approach to synthesis magnetic metal organic
frameworks (MMOFs) for methylene blue (MB) adsorption from waste water (Fig.4). Fe 3O4
microspheres are used as the magnetic cores in this material, and they also provide Fe (III)
for the synthesis of MIL-100(Fe), which is grown as a shell on the surface of the Fe 3O4. The
thickness of the shell can be controlled by adjusting the reaction time. The Fe 3O4@MIL-
100(Fe) core-shell structure has significantly increased surface area and pore volume while
maintaining its magnetism, making it an effective adsorbent for removing dyes such as
methylene blue (MB) from water. The MIL-100(Fe) is uniformly grown as a shell on the
surface of Fe3O4, and the shell thickness can be fine-tuned from 73.5 to 148 nm by
controlling the reaction time. The surface area and pore volume of Fe 3O4 was measured to be
26 m2 g-1 and 0.0483 cm3 g-1 respectively. The Fe3O4@MIL-100(Fe) core-shells showed high
adsorption capacity (221 mg g-1) towards methylene blue (MB) dye, based on electrostatic
interactions and size filter effect [27].

Figure 2:Schematic representation of in situ one-step hydrothermal synthesis of Fe 3O4@MIL-

100(Fe) core-shell magnetic microspheres [27]

Magnetic and porous Ni@MOF-74(Ni) composite obtained by one-step solvothermal

reaction of Raney Ni and DOBDC. The Ni@MOF-74(Ni) composite exhibits both magnetic
characteristics and high porosity, enabling it an efficient candidate for dye removal and target
drug delivery system. The solvent desorption was tested to evaluate the feasibility of
regeneration the RhB saturated Ni@MOF-74(Ni). The Ni@MOF-74(Ni) composite after
adsorption reached to equilibrium, was separated by magnet and washed with eluent to
evaluate its reusability. The adsorption amount of rhodamine b reached up to 177.8 mg g −1,
and 4.1 mg g−1 for ibuprofen. The Ni@ MOF-74(Ni) holds great potential as sorbent for
targeted drug delivery [28].

The Pt@DUT-5 nanocomposite was obtained by one-step approach for the incorporation of
tiny metal nanoparticles (MNPs) within metal-organic frameworks (MOFs) without
additional stabilizing agents and reductants. The method involves directly mixing the
precursors hexachloroplatinic (IV) acid and MOFs in DMF. Here, the solvent DMF plays
dual role one as solvent in the synthesis of various MOFs and other hand as reducing agent
under suitable conditions for the fabrication of metal nanoparticles with uniform shape and
size. The resulting Pt@DUT-5 nanocomposites exhibit excellent stability and significantly
enhanced catalytic activity and selectivity as compared to the commercial Pt/C in liquid-
phase aerobic oxidation of alcohols [29].
The advantage of in situ one-step synthesis is that it is a simple and efficient method for
synthesizing core-shell structures. It eliminates the need for multiple steps and reduces the
risk of contamination. The disadvantage is that it may be difficult to control the size and
thickness of the shell, which can affect the properties of the core-shell structure [29].
2.2.3. Layer by layer Strategy
Layer-by-layer (LBL) deposition is a well-established method of fabricating nanosized
MOFs, in which metal secondary building units (SBUs) and organic linkers are deposited
onto the substrate surface stepwise [30]. The layer-by-layer method is particularly useful for
creating smooth and continuous layers of MOFs on planar substrates. However, on high-
curvature substrates of high surface energy, such as surfaces of nanoneedles and
nanoparticles, the nanosized MOF crystallites may not grow into smooth and continuous
layers. This can result in more cracks and defects, which can make more surface sites
available and improve the catalytic efficiency of the MOFs. Moreover, if magnetic
nanoparticles are used as substrates for LBL growth, magnetism can be easily integrated into
the nanosized MOFs. This can facilitate their recovery after catalysis and make MOFs more
suitable for intended application [30], [31].
The Fe3O4@PDA-Pd@[Cu3(BTC)2] catalyst was prepared using a facile and versatile
strategy. The strategy involves the synthesis of Fe3O4 NPs, followed by the formation of a
polydopamine (PDA) layer on the surface of Fe 3O4 NPs, and then the deposition of Pd NPs on
the PDA layer. Finally, the Pd-decorated Fe 3O4@PDA was coated with a layer of Cu3(BTC)2
MOF. The resulting catalyst showed high surface area than (126.28 m 2g -1
), pore size
(4.79nm), and magnetization strength (23.29 emu g -1) [32].

Figure 3:Schematic Illustration for the Layer-by-Layer Fabrication of Core−Shell Fe 3O4@PDA-

Pd@[Cu3(BTC)2] [32]

The process of layer by layer can be time-consuming and requires multiple steps. This
process can also be quite sensitive to environmental conditions such as temperature and
humidity, which can further complicate the task at hand. However, there are some significant
advantages to this method that make it worth considering. Firstly, it allows for the precise
control of the thickness and composition of the deposited layers, which is incredibly
beneficial for those looking to create highly specific structures. Additionally, this method can
be used to create complex and multifunctional structures, which adds to its versatility.
Finally, the resulting structures can have unique properties and applications due to the
precision with which they are created. Overall, while the process of layer by layer may be
challenging, the benefits it offers make it a valuable tool in certain situations [32].
2.2.4. Embedding Method
The embedding technique involves the attachment of magnetic particles onto the surface of
MOFs. When using the embedding method to create magnetic frameworks composite
(MFCs), the magnetic nanoparticles (MNPs) are typically added into a mixture of MOFs that
promote nucleation and growth processes. Afterwards, the MNPs are embedded into the
MOFs material formed through ultrasonication or hydrothermal conditions. In general, the
structure of MFCs synthesized using the embedding method closely resembles that of MOFs
[33].

Fe3O4/ZIF-8 was synthesized via embedding method, the process involves synthesize
Fe3O4/ZIF-8 nanoparticles at a room-temperature reaction between 2-methylimidazolate and
zinc nitrate in the presence of Fe3O4 nanocrystals. The particle size, surface charge, and
magnetic loading can be controlled by adjusting the dosage of Zn (NO 3)2 and Fe3O4
nanocrystals. The resulting nanoparticles have good thermal stability, large surface area, and
a superparamagnetic response, which allows them to be easily separated and dispersed using
an external magnetic field [33].

Figure 4:Schematic illustration for the formation mechanism of Fe3O4/ZIF-8 particles [33]

The embedding method has several advantages. Firstly, the steps involved in this method are
relatively simple which simplifies the overall process. Additionally, MOF growth is
completed in a single step which makes it more efficient. Finally, the embedding method is
applicable to the magnetization of most MOFs which increases its versatility. However, it is
important to note that this method also has certain limitations. It can be challenging to
prevent direct nucleation and growth of MOFs in solution. Moreover, a part of the MNPs
often gets embedded in the pores of the MOFs, which ultimately leads to a reduction in the
specific surface area [33].
2.2.5. Encapsulation
The method of encapsulation chiefly involves employing a buffer interface to link the porous
frame with the MNPs, so as to encourage MOFs growth around the MNPs for manufacturing
MFCs. The MNPs are initially pre-incorporated into the polymer layer and then added to the
nucleation and growth mixture of MOFs to enhance the compatibility of MFCs. This
technique is frequently employed for creating composite materials with MNPs serving as the
core.
The Fe3O4@SiO2@UiO-66 core-shell microspheres were created using the encapsulation
process. Initially, the Fe3O4@SiO2 microspheres underwent modification with glutaric
anhydride to create carboxylate-terminated Fe 3O4@SiO2 microspheres, which served as the
base for UiO-66 growth under hydrothermal conditions (Fig. 11). Microsphere images
obtained via TEM and SEM display that UiO-66 crystals formed a continuous framework
that encapsulated the Fe3O4@SiO2 microspheres [34].
Figure 5:Schematic of encapsulation for the synthesis of Fe 3O4@SiO2@UiO-66 core-shell
microspheres [34]

The encapsulation method presents some advantages, such as relatively simple operation
steps and the possibility of obtaining MFCs with a core-shell structure by controlling the
reaction conditions or modifying the MNPs. Nonetheless, this methodology also has some
limitations that must be considered. For instance, in order to enhance the compatibility of
MFCs materials, MNPs must be pre-embedded into the polymer layer. Additionally, it is not
an easy task to achieve uniform MOF coating, which represents another challenge for the
encapsulation method [34].

2.2.6. Direct Mixing Method

The method of creating MFCs involves the use of ultrasonic or high-temperature
polymerization to uniformly mix previously prepared MNPs and MOFs. This attachment
allows for the MFCs to be retrieved from the sample matrix via external magnets. The
primary objective of this method is to regulate the interaction between magnetic materials
and MOFs to establish a durable bond. Hu et al. reports facile and efficient strategy for the
synthesis of a novel kind of hybrid magnetic metal-organic framework (MOF)-5 via chemical
bonding assembly. The covalent bonding established between the amino functionalized Fe 3O4
nanoparticles and the surface of the metal organic framework improved the chemical stability
and structure uniformity of the hybrid microcrystals. Combination of MOF-5 with Fe 3O4
nanoparticles allows for facile withdrawal of the porous materials by magnetic
decantation [35].
Figure 6:Schematic of the fabrication processes of the hybrid magnetic MOF-5 [35]

The direct mixing procedure is a simple and convenient method used in the preparation of
Metal Framework Composites or MFCs. It has numerous advantages such as its ease of use
and its applicability to any MOF. Despite these advantages, it does have a few limitations that
need consideration. Firstly, MNPs used in this approach are not firmly bound to MOF
meaning that they can easily detach from it. Secondly, this method requires a relatively large
amount of MOF which can be quite expensive depending on the acquisition cost of the
selected MOFs [35].

2.2.7. Self-Template Method

Self-template synthetic approach refers to a method of creating a material where a precursor material
is used as a template for the final product. In case Fe 3O4@SiO2@HKUST-1 core–shell nanostructures,
Cu (OH)2 was used as a self-template to create the HKUST-1 shell in the magnetic MOF composite,
followed by conversing into HKUST-1 at room temperature in water-ethanol mixture solvent to
obtain Fe3O4@SiO2@HKUST-1 core-shell nanostructures. This approach allows for precise control
over the final product's structure and properties [36].

Figure 7:Schematic diagram of the preparation of Bi-I-functionalized magnetic HKUST-1

composites [36]
The self-template method has advantages such as it is a simple and green method, it does not
require any additional templates, and it can be used to prepare various types of MOF
composites. However, it also has some disadvantages such as the uncontrollable growth of
the MOF crystals, which can lead to the formation of large particles and the loss of the
magnetic properties of the composites [36].

2.2.8. Dry Gel Conversion Strategy

Dry gel conversion (DGC) is a method used to synthesize magnetically responsive
MOF/Fe3O4 composites. In study of synthesis of HKUST-1/Fe3O4 composites, DMF vapor is
generated at an elevated temperature and diffuses to contact with the parent materials, leading
to the growth of HKUST-1 in a confined environment. As a result, HKUST-1 is forced to
combine with magnetic Fe3O4 nanoparticles. The HKUST-1/Fe3O4 composites, where
magnetic Fe3O4 nanoparticles are incorporated into the HKUST-1 matrix, are successfully
fabricated by this strategy. DGC simplifies synthetic procedures and minimizes solvent
consumption [37].

Figure 8:Schematic diagram for the synthesis of (a) HKUST-1 by DGC, and the HKUST-1/Fe 3O4
composites by (b) DGC and (c) the solvothermal method [37]

The method has certain advantages such as the prevention of incorporation of magnetic
nanoparticles into pores of framework structures and the reduction of organic solvent losses.
However, it also has some limitations. One of these limitations is the difficulty in controlling
the uniformity of growth of the metal-organic framework structure [37].
 2.2. Applications of Magnetic framework composites in Enzymatic
Hydrolysis
Magnetic framework composites (MFCs) have been extensively studied for their potential in
various applications including catalysis, separation, and sensing. In particular, MFCs have
shown promising results in enhancing the efficiency of hydrolysis reactions. Hydrolysis is a
chemical reaction that involves the cleavage of chemical bonds through the addition of water
molecules. This reaction is commonly used in various industrial processes such as food
processing, pharmaceutical production, and biofuel production. MFCs are composed of a
magnetic core, which allows for easy separation from reaction mixtures using an external
magnetic field, and a porous composite shell, which provides a high surface area for catalytic
activity. The composite shell can be functionalized with various metal ions or organic groups,
which can further enhance the catalytic activity and selectivity of the material. At present,
enzymatic hydrolysis utilizes magnetic composite frameworks that function by confining
enzymes capable of breaking down specific substances. This immobilization aids the
hydrolysis process and enhances enzyme stability and reusability. Several materials have
been used as carriers for immobilizing enzymes, such as polymers, nanoparticles, porous
materials, and microcapsules. While using these materials can enhance the stability of
enzymes in extreme conditions, there are still problems such as enzyme leaching,
denaturation, and limited mass transfer efficiency that may cause a reduction in the
effectiveness of the enzymes over time, but they cannot guarantee their effective recovery
from the reaction medium. Magnetic frameworks composite can give recovery, stability and
selectivity to the immobilized enzyme.
Yuan Ji et al. reports a mild and quick one-pot strategy to encapsulate lipase and Fe 3O4
nanoparticle into ZIF-8 to form CRL/MNP@ZIF-8 with 88.4% immobilization yield [38]. The
resulting magnetic MOF composite demonstrated an excellent protective capability against
harsh environments and could also simplify the complex downstream separation and recovery
operations. The residual CRL/MNP@ZIF-8 still retained 82% activity after five successive
cycles of reuse, which can significantly reduce the processing cost and might be applicable in
industries in the future. One limitation is that the authors did not compare the performance of
their CRL/MNP@ZIF-8 composite with other enzyme immobilization methods. Additionally,
the they did not investigate the effect of different reaction conditions on the performance of
the composite [38].
Also, Sumanta et al. reported a one-step in situ hydrothermal route to construct MOFs
encapsulating superparamagnetic Fe3O4 nanoparticles [39]. A one-step in situ hydrothermal
approach for the fabrication of MOFs encapsulating superparamagnetic Fe 3O4 nanoparticles
is established. The incorporation of Fe 3O4 nanoparticles into MOFs results in a variety of
intriguing intrinsic features, including porous nature, ease of functionalization, and
significant superparamagnetic. Candida Rugosa Lipase (CRL) was covalently attached to
amino-rich magnetic MOFs. The loading amount of CRL on magnetic MOFs was 280 mg/g
support and the immobilized CRL was efficiently recycled up to nine cycles. The
immobilized CRL was efficiently recycled up to nine cycles. The activities of lipase were
compared with the activity of the first running using hydrolysis of p-NPB. One limitation is
that they did not investigate the effect of the size of Fe 3O4 nanoparticles on the properties of
the resulting magnetic MOFs. Additionally, they did not compare the performance of their
magnetic MOFs with other reported MOFs for lipase immobilization [39].
Chaoping et al. introduces the fabrication of a novel magnetic hierarchically porous metal-
organic framework (MOF) using a modulator-induced defect-formation strategy [40]. The
MOF was used to immobilize amidase, resulting in a biocomposite with superior enzymatic
catalysis performances compared to the free enzyme and its counterparts without hierarchical
porosity. The morphology and structure of the immobilized amidase remained relatively
intact after 15 cycles and last long-time storage. The magnetic hierarchically porous MOF
shows competitive catalytic activity, enhanced thermal stability, organic solvent tolerance
and storage stability. The enzyme loading capacity of the hierarchical Fe 3O4@MOF was
increased up to 211.3 mg g -1 using glutaraldehyde (GA) via covalent cross-linking. The
immobilized amidase on magnetic hierarchically porous MOF shows competitive catalytic
activity, enhanced thermal stability, organic solvent tolerance and storage stability compared
to the free enzyme and its counterparts without hierarchical porosity. One limitation is that
they did not investigate the effect of different GA concentrations on the immobilization
efficiency and activity of the immobilized amidase. Additionally, they did not compare the
performance of the immobilized amidase with other immobilization methods [40].
Wang et al. synthesized immobilized alpha-L-rhamnosidase enzyme on magnetic metal-
organic frameworks (MOFs) using the cross-linking agents EDC/NHS [41]. The immobilized
enzyme Rha@MOF reached an activity of 25.09 U/g with a lower apparent Km value
compared with the free enzyme. The conversion rate of 20 g/L rutin was 91.42%,
corresponding to an iso-quercitrin productivity of 12.78 g/L/h. Rha@MOF also exhibited
significantly improved reusability; the conversion rate was still 73.55% after 30 cycles at
60◦C. The immobilized alpha-L-rhamnosidase enzyme on magnetic metal-organic
frameworks (MOFs) using the cross-linking agents EDC/NHS showed improved reusability
and conversion rate, making it a feasible biocatalyst for the conversion of flavonoids with
low aqueous solubility. But they did not compare the performance of their immobilized
enzyme with other immobilization methods or with other enzymes that can hydrolyze rutin.
Additionally, the authors did not investigate the stability of the immobilized enzyme under
different storage conditions. One limitation of this paper is that they did not compare the
performance of their immobilized enzyme with other immobilization methods or with other
enzymes that can hydrolyze rutin. Additionally, they did not investigate the stability of the
immobilized enzyme under different storage conditions [41].
Yinhua. et al. successfully immobilized of cellulase on a magnetic core-shell metal-organic
framework (MOF) material, UIO-66-NH2 [42]. The immobilized cellulase showed better
tolerance to inhibitors and easier recycling compared to its free form. The study also found
that immobilization can alleviate the inhibitory effects of certain pre-treatment inhibitors on
cellulase. Immobilized cellulase could be recycled for up to 5 consecutive runs. Furthermore,
compared to free cellulase, immobilized cellulase showed better tolerance to formic acid and
vanillin, two typical inhibitors found in lignocellulosic prehydrolysates. In the presence of 5
g/L of formic acid and vanillin, immobilized cellulase demonstrated 16.8% and 21.5% higher
activity than free enzyme, respectively, and its improvement in hydrolysis yield was 18.7%
and 19.6% respectively. One limitation of this study is that they only evaluated the
performance of immobilized cellulase in terms of stability, efficiency, and tolerance to
inhibitors. The study did not investigate the mechanism behind the improved performance of
immobilized cellulase or the effect of immobilization on the structure and function of
cellulase. Additionally, the study only used one type of MOF material for immobilization,
and it is unclear whether other MOF materials can also improve the performance of
immobilized cellulase [42].
To summarize, The MFCs are a combination of MNPs and MOFs materials that offer the
benefits of both. They possess the magnetic features of MNPs and the catalytic performance,
selectivity, and high adsorption of MOFs. They have a large specific surface area and more
active sites on their surfaces than traditional porous materials. They can be retrieved quickly
and simply using external magnets, thereby reducing processing time. The future prospect of
magnetic frameworks composite is promising as it has shown high catalytic activity towards
the hydrolysis of chemical warfare agent simulants.
The core-shell magnetic metal-organic frameworks (MOFs) have a well-defined structure,
good thermal and chemical stability, and rich mesoporous structure. The magnetism is
retained after the coating of MOFs, which makes it easy to recover and reuse after catalysis.
The stability of the core-shell structure has been verified through an 'interrupt' experiment,
which showed that the effective catalytic MOF component was completely removed along
with the separation of the magnetic particles. This indicates that the core-shell structure is
stable and does not easily fall off under stirring conditions.
Overall, despite significant progress, several challenges hinder the commercial viability of
cellulose hydrolysis. These include high enzyme costs, low reaction rates, inhibitor
formation, and limited understanding of reaction mechanisms. Future research directions
include the development of efficient catalysts, optimization of reaction conditions, and the
exploration of innovative technologies such as biocatalytic cascades and combined
hydrolysis-fermentation processes. Moreover, process integration, techno-economic analysis,
and environmental considerations are crucial for the sustainable production of valuable
chemicals from cellulose.
3. Methodology

3.2. Materials and Equipment’s  Thermometer

 Teflon line Autoclave  Heating mantle
 Beakers  Glass funnel and filter paper
 Sample holders  Erlenmeyer flask
 Electrical balance  FTIR
 Magnetic stirrer/hotplate  TGA
 pH meter  X-ray diffractometer
 Vacuum pump and Buchner  Magnetometer
funnel
 Uv-visible spectrophotometer
3.3. Chemical and Reagents
 FeCl3.6H2O
 α- Cellulose
 Sodium citrate
 Mercapto-acetic acid
 Sodium acetate
 Ethylene glycol
 Monocarboxylic acids
 CuSO4 (98%)
 Terephthalic acid
 N, N-dimethylformamide (DMF)
 Deionized water
 NaOH pellets
 HCL
 Ethanol
 3.4. Experimental Methods

3.4.1. Synthesis MOFs

The Cu-MOFs will be synthesized by mixing copper sulfate and terephthalic acid in N, N-
dimethylformamide (DMF) solvent at a certain temperature and pressure via ultrasonication.
The mixture will be then sealed in a Teflon reactor and maintained at for certain temperature
and time. After that the reaction will be cooled down at room temperature, the precipitate will
be recovered by centrifugation and washing with DMF for several times. Then the solids will
be suspended in methanol overnight. Finally, the resulting powder was dried at certain
temperature and time [43].

3.4.2. Synthesis of HP-MOFs by Modulate defect Induced Strategy

CuSO4 and excess amounts of modulator (acetic acid or formic acid) first will be dissolved in
DMF in a vial to give Cu-oxo clusters by ultrasonication for certain time, then BDC will be
added. After the formation of uniform solution by ultrasonication, two vials of mixture
solution will equally divide into three Teflon-lined autoclave which will be allowed to react
at set temperature in oven for days, then the precipitate in 3 autoclaves will be recovered by
centrifugation and will be washed with DMF for three times. To activate the samples and
further remove the residual alkyl acids inside Cu-MOF, the obtained powder will be treated
in the mixture solution of concentrated HCl and DMF at certain temperature and time. After
repeating this treatment, the product will be centrifuged and washed with DMF and methanol
for several times. After suspending in methanol overnight, the resulting powder will be
evacuated in a vacuum oven at certain temperature and time [17].
3.4.3. Synthesis of magnetic HP-MOFs
3.4.3.1. Synthesis of functionalized Fe3O4
Particles of Fe3O4 will be synthesized via a hydrothermal method by mixing, FeCl 3.6H2O,
sodium citrate and Sodium acetate were dissolved in ethylene glycol with magnetic stirrer.
Then the mixed liquor will be transferred to a sealed Teflon-lined stainless-steel to be heated
in an autoclave. The resultant Fe3O4 will be collected, washed with ethanol several times, and
dried under vacuum. Thioglycolic acid functionalized Fe3O4 nanoparticles will be prepared
which can provide a stable coating and enhanced solubility in water. Fe 3O4 will be added to
ethanol solution of thioglycolic acid under shaking. The product was collected by an external
magnetic field and washed [44].
 3.4.3.2. Synthesis of magnetic HP Cu-MOF
The magnetic properties of the Cu-MOFs will be imparted by adding magnetic nanoparticles
during the synthesis process. A gram of functionalized Fe3O4 microspheres will be dispersed
in ethanol of containing gram of CuSO4 and will be sonicated for set time. Then, solution of
terephthalic acid will be added into the above solution at a constant rate followed by
continuous mechanical stirring. The resulting Fe3O4-@MOF microspheres were collected
from the reaction mixture under an external magnetic field, washed with ethanol several
times, and dried at under vacuum [45].
3.4.4. Hydrolysis of cellulose using Fe3O4-@HP-MOF
Pretreatment of cellulose in a hydrothermal reactor that is will be heated in an oven as
previously developed method [24]. Specifically, we will soak gram of cellulose powder in
DI-water with or without Fe3O4-@HP-MOF. Then, the mixture will be heated at different
temperature for different time independently. Finally, the mixture to will cool down to room
temperature and filtered by magnetic decantation using external magnetic field. Wash the
solids with distilled water and dry them in a vacuum oven. Record the weight of the solid
obtained and measure the degree of conversion of cellulose [24].
3.5. Characterization of magnetic Cu-MOFs
The physical and chemical properties of the synthesized magnetic Cu-MOFs will be
characterized using various techniques. XRD will be used to determine the crystal structure
and phase purity of the synthesized materials. SEM and particle size of the synthesized
materials. FTIR spectroscopy will be used to identify the functional groups present in the
synthesized materials. Magnetization measurements will be used to determine the magnetic
properties of the synthesized magnetic Cu-MOFs.
3.6. Evaluation of catalytic performance
The catalytic performance of the synthesized magnetic Cu-MOFs in cellulose hydrolysis will
be evaluated by conducting hydrolysis experiments using different substrates cellulose. The
hydrolysis experiments will be carried out in the presence of the synthesized magnetic Cu-
MOFs and non-magnetic Cu-MOFs under different reaction conditions like temperature,
time, and catalyst loading. The hydrolysis products will be analysed using various analytical
techniques with HPLC. The catalytic performance of the synthesized magnetic Cu-MOFs will
be compared with that of non-magnetic Cu-MOFs to evaluate the effect of magnetic
modification on the catalytic activity.
 3.7. Investigation of reusability and stability
The reusability and stability of the magnetic Cu-MOF catalysts will be investigated by
conducting multiple cycles of cellulose hydrolysis using the same catalysts. The catalysts will
be recovered by magnetic separation and washed with solvents to remove any remaining
residues. The recovered catalysts will be characterized using various techniques to determine
their stability and changes in physical and chemical properties after each cycle of catalysis.
The catalytic performance of the recovered catalysts will be evaluated in subsequent cycles of
biomass hydrolysis to determine their reusability. In general, the synthesis of magnetically
modified Cu-MOFs for biomass hydrolysis is a promising area of research with significant
potential for industrial applications. The proposed research aims to synthesize and
characterize magnetic Cu-MOFs and evaluate their catalytic performance and reusability in
biomass hydrolysis. The expected outcomes of this research can contribute to the
development of efficient and sustainable processes for the production of biofuels and
chemicals from lignocellulosic biomass.
4. Budget Breakdown
Table 1: Total expenses

Activities Cost (Birr) Remark

Chemical and Materials cost 4000 Table 3
Printing and binding cost 9000 Table 4
Labor and Characterization costs 12,000 Table 5
Grand Total (Birr) 25,000

Table 2:Chemicals and materials cost

Item Unit Qty Unit Price (Birr) Sum of costs (Birr)

Terephthalic acid 100g 1 500 500
Thioglycolic acid 25ml 1 900 900
Cellulose powder 100g 1 600 600
Sodium Citrate 100g 1 400 400
Sodium Acetate 100g 1 300 300
Formic acid 100ml 2 1300 1300
Total Cost (Birr) 4000

Table 3:Printing and binding cost

Item Unit Qty Unit Price Sum of costs (Birr)

(Birr)
Printing Pages 1000 5 5000
Binding Number 3 600 2400
Flash 32 GB 1 600 600
CD 4 GB 10 100 1000
Total Cost (Birr) 9000

Table 4: Labor and Characterizations cost

 Item Fee No. Total fee (Birr)
(birr/day)
Labor and characterization 600 20 6000
Total Cost (Birr) 12,000

5. Work Plan
Table 5: Timeline for the proposed research work

Activities Time schedule

May. 2023 Jun.2023- Au.2023 Oct.2023- Dec. 2023
Jul.2023 Sep.2023 Nov.2023
Chemical and material
collection
Experimental work
Characterization part
Application Part
Result analysis, Manuscript
Preparation and Final report
preparation
Thesis Defence According to University schedule