Enamala2018 PDF

Renewable and Sustainable Energy Reviews 94 (2018) 49–68
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Renewable and Sustainable Energy Reviews

journal homepage: www.elsevier.com/locate/rser
Production of biofuels from microalgae - A review on cultivation, T

harvesting, lipid extraction, and numerous applications of microalgae
Manoj Kumar Enamalaa, Swapnika Enamalab, Murthy Chavalic, Jagadish Donepudid,
Rajasri Yadavallie, Bhulakshmi Kolapallia, Tirumala Vasu Aradhyulaf, Jeevitha Velpurig,
⁎
Chandrasekhar Kuppamh,
a
Acharya Nagarjuna University, Guntur, Andhra Pradesh, India
b
GSL Medical College, Rajahmundry, Andhra Pradesh, India
c
MCETRC, 20-26-136, Chiravuru, Tenali, 522201 Guntur, Andhra Pradesh, India
d
Mechanical Engineering Department, Narasaraopeta Engineering College, Narasaraopet, Guntur, Andhra Pradesh, India
e
Sreenidhi Institute of Science and Technology, Yamnampet, Ghatkesar, Telangana, India
f
Department of Mechanical Engineering, National Chung Cheng University, Taiwan
g
Environmental Science and Technology, JNTUH, Hyderabad, Telangana, India
h
Green Processing, Bioremediation and Alternative Energies Research Group, Faculty of Environment and Labour Safety, Ton Duc Thang University, Ho Chi Minh City,
Vietnam
A R T I C LE I N FO A B S T R A C T
Keywords: The concern regarding alternate sources of energy is mounting day-by-day due to the effect of pollution that is
Bioenergy damaging the environment. Algae are a diverse group of aquatic organisms have an efficiency and ability in
Algae biomass mitigating carbon dioxide emissions and produce oil with a high productivity which has a lot of potential
Harvesting procedures applications in producing biofuel, otherwise known as the third-generation biofuel. These third generation
Lipid
biofuels are the best alternative to the present situation since they have the perspective to eliminate most of the
Biodiesel
Third generation biofuel
ecological problems created by the use of conventional fossil fuels. These organisms are responsible for closely
50% of the photosynthesis process taking place on the planet and are distributed predominantly in many of the
aquatic systems. The huge interest in utilizing these organisms as a potential source of energy lies in converting
the primary as well as secondary metabolites into useful products. Algae are considered to be the most prominent
resource for the upcoming generations as the most suitable and sustainable feedstock. The key process limita-
tions in microalgal biofuel production are inexpensive and effective harvesting of biomass and extraction of
lipids. The major objective of this article is to provide a comprehensive review on various methods of both
biomass harvesting and lipid extraction from microalgae available, so far, besides to discuss their advantages and
disadvantages. This article also deals with various conditions that are favourable for lipid accumulation as well
as the yield from different species.
1. Introduction major disadvantage [3,4]. The greenhouse gas (GHG) levels in the en-
vironment have increased at an alarming rate in the post-in-
The energy crisis is increasing globally due to the heavy industrial dustrialization era by 25% of the total [5]. Natural causes, as well as
development and exponentially growing population. Sources like human activities, have been mentioned as the major causes of this rise
petrol, diesel, natural gas, coal which were considered to be the basic in temperature leading to global warming [1]. The major contributors
sources for fuelling the life are getting exhausted due to extensive usage include carbon dioxide (CO2), methane (CH4), nitrous oxide (N2O), and
[1,2]. Moreover, these fossil fuels release a lot of toxic and harmful other fluoro-hydrocarbons. Among them, the major pollutant which
gases into the atmosphere and pollute the environment which is the damages the environment is CO2 [1,6]. The above-mentioned gasses are
Abbreviations: GHG, Greenhouse gas; N2O, Nitrous oxide; TCA, Tricarboxylic acid; ETC, Electron transport chain; ATP, Adenosine triphosphate; Glu-6-P, Glucose-6-phosphate; PPP,
Pentose phosphate pathway; NH4+, Ammonium; NH3, Ammonia; O-U, Ornithine-urea; WC, Water column; WB, Water bodies; NaSO4, Sodium sulphate; DIC, Dissolved inorganic carbon;
FeCl3, Ferric chloride; Fe2SO43, Ferric sulphate; CaOH2, Calcium hydroxide; MgOH2, Magnesium hydroxide; R-NH2, Amine groups; Al2SO43, Aluminium sulphate; bio-CH4, Bio-methane;
bio-H2, Biohydrogen; FA, Fatty acids
⁎
Corresponding author.
E-mail address: [email protected] (C. Kuppam).
https://doi.org/10.1016/j.rser.2018.05.012
Received 9 May 2017; Received in revised form 12 May 2018; Accepted 13 May 2018
1364-0321/ © 2018 Published by Elsevier Ltd.
M.K. Enamala et al. Renewable and Sustainable Energy Reviews 94 (2018) 49–68
Fig. 1. Classification of algae explained in a simple way.
present in the atmosphere at a normal rate but due to the emissions The reasons for algae being the preferred source over plant sources
from the vehicles, their concentration has increased over the past few
decades [7]. Owing to all this, there has been a change in climatic (a) The microalgae have a high efficiency for photosynthesis with an
conditions over the globe, which has become a topic of debate [8]. At adaptability to a wide range of light and temperature variations
this time, replacing fossil fuels with other alternative sources especially [16].
those that benefit the environment is the best solution [1,4]. These (b) The microalgae can grow in water with different levels of nutrients
microalgae sources act as solar driven energy cell factories and are and can adjust to the change in the growth characteristics and
capable of converting CO2 to oxygen (O2) and thus reducing the toxic nutrient uptake ability [16].
substances and chemicals in the environment. Hence these organisms
are very promising in this aspect [9]. These organisms have a larger surface to volume ratio, which en-
The working machinery of these organisms is the same as that of the ables them to grow very efficiently. Fixation of CO2 at different water
plants, as both are photosynthetic. These utilize the sunlight from the levels is achieved very easily. But the major challenge would be the
atmosphere for the photosynthesis process and other essential nutrients cultivation of microalgae on a large-scale, harvesting and finally con-
from the surroundings for their growth [10]. There are also many food verting into useful fuels which are beneficial for the human society and
crops available which are used for the production of fuel apart from as well have an economic impact [12].
algae. Much of the study is being carried out on industrial production of In this review article, we have discussed the state-of-the-art in
biodiesel from plant sources. Apart from soya bean oil, jatropha, left- biofuel production from microalgae. The distinctiveness of this review
over cooking oil, canola, corn, and animal fats etc., are also being tried is in its coverage of numerous harvesting procedures, extraction
as fuel sources [11]. However, these sources must also meet the re- methods and parameters which are involved in growth and the lipid
quirement for the food for human beings. Upon extensive usage of these extraction techniques. In this paper, we have given a tabular column
sources for oil production, there may arise a scarcity in providing food and illustrated various conditions that favor lipid accumulation as well
for human beings [11]. Production of biofuels from the plant sources as the yield from different species. We also discuss various maximum
was criticized by many scientific communities as well as local farmers growth rate values, lipid percentage accumulated in their cells of in-
and the general public since the growth of these plants needs an ex- dividual species, numerous methods regarding biofuel and co-products
tensive usage of land, leading to a crisis in food grain production. recovery, carbon dioxide mitigation and wastewater treatment.
The biofuels are divided into three generations depending on the
source from which they are obtained [12]. 1.1. General characteristics of algae
• First generation biofuels derived from plant sources. The green algae and the cyanobacteria together called the blue-
• Second generation biofuels derived from agricultural wastes, lumber green algae consist of a huge group of photosynthetic organisms, the
wastes etc. most efficient organisms reported to date. Unlike other microorganisms,
• Third generation biofuels derived from microalgae. these have abundant chlorophyll inside the cells, with a well-defined
nucleus, cell wall, and pigments [8].
Researchers have turned their interest towards fuel production from
one of the oldest living creatures on the earth, microalgae. These are 1.2. Forms of algae
utilized not only in producing fuels but also in capturing the CO2 from
the atmosphere which helps in cleaning the environment and producing The various forms of algae which exist are:
better air to breathe [13,14]. There are two different classes of algae
known as macroalgae and microalgae. These photosynthetic organisms • Colonial, Capsoid, Coccoid, Palmelloid, Filamentous, Parenchymatous.
are mainly found in aquatic habitats both freshwater and marine. These
are microscopic and have very amazing and fascinating structures [15]. The cell walls of diatoms comprise polymerized silica known as a
50
Fig. 2. Food web of algae which shows the relation between humans and other species for its survival.
Table 1
Advantages and disadvantages of microalgae.
Advantages Disadvantages
• Very short doubling time • They can easily grow in any aquariums
• Cheap media can be used (including wastewater) • When algae are attached to a system it produces methane which gets mixed with the water sources.
• Can be supplied as a food for aquaculture • Risk in culturing pure cultures of algae due to the bacterial contamination
• Absorbs CO as it grows and helps in cleaning of the environment • High temperatures are a serious threat to open pond cultivation system
• Can • Due
2
be grown in an non-arable land on a large scale and on small to the large growth of algae on the surface of lakes, ponds they obstruct the light to reach the
scale can be grown in our own houses aquatic plant's fishes and other aquatic species which are deep under water.
• The great decrease of competency for food vs fuel.
• water).
New source of fuel can be obtained (capability to produce H from
2
• Algae contain a high amount of iron (Fe) which is advantageous if

consumed by the pregnant women.
• Wastewater can be purified
• Contains rich lipid profile which can be used for improving health
• Can be grown in open pond cultivation system to several
kilometers
• They are considered as tough competitors for biofuel production
• Solar conversion increases by 10 folds when compared to plants
• Cyanobacteria can fix N from the atmosphere
2
frustule. The diatoms often accumulate oils and chrysolaminarin [16]. encapsulated vacuoles when dissolved in water and is stored in the cells
The green algae are particularly rich in fresh water (Fig. 1). They pro- [18]. The detailed information regarding advantages and disadvantages
duce starch as major chief storage compound by the photosynthesis of microalgae are provided in Table 1.
mechanism. However, they can also produce fats and oils. The freshwater
green algae Haematococcus pluvialis are a freshwater species of chlor- 2. Metabolism of algae
ophyte, which is a chief source of strong antioxidant ‘astaxanthin’, which
is very significant in aquaculture, and cosmetics industry. Hence it is The metabolic reaction process is almost same in all the photo-
having high commercial importance [16]. The Chrysolaminarin consists synthetic organisms. The most important factor is the nutrition uptake
of a linear polymer arranged in the chains of β(1−3) also β(1−6) linked from the surroundings through various biochemical and transportation
glucose molecules (in 11:1 ratio), which was earlier well-known as leu- process [17]. The carbon (C) and nitrogen (N) are considered to be the
cosin. The Chrysolaminarin is considered as storage polysaccharide as important elements in the photosynthetic metabolic pathways. The
well as the most common biopolymer in the world [17]. It is also utilized major changes which occur during the metabolic pathways are the mass
as a reserve food by organisms such as Bacillariophyta which is similar to of the cells, volume, densities, protein, chlorophyll, RNA, and vitamin
the laminarin of brown algae (Fig. 2). The Chrysolaminarin exists as an contents [19,20].
51
2.1. Carbon metabolism the C-rock formation and for generating the reserves for the fossil fuels
[28]. When compared with the ancient algae, the modern algae produce
The C metabolism starts with the incorporation of glucose into the half of the oxygen like which the plants and are also responsible for
algal cells and the addition of phosphate group to hexose which yields cycling of major elements such as sulfur (S), phosphorous (P), C, N and
glucose-6-phosphate (Glu-6-P), which is easily accessible for the sto- other trace elements. Hence in nature also these elements play a vital
rage, growth, and respiration in the cells [21]. In the darker conditions, role in various interactions and controlling the atmospheric conditions
algae cannot metabolize glucose because a short supply of energy is [26].
expelled through dissimilation of glucose [21]. However, the in- Like many of the other microorganism’s algae have also played a
sufficient amount of the enzyme lactate dehydrogenase will be slowed major role in shaping up with the earth's biogeochemistry and in the
down this process. Nielsen and Lewin have shown that only Embedded coming days these activities would be similar or may be higher and can
Mayerhof and Pentose phosphate pathway (PPP) have been shown in be compared with the human activities [29]. Moreover 99.9% of the
the algal cells [22]. These algal cells utilize almost the entire glucose total biomass of the algae is accounted by 6 main elements such as C, N,
present in the environment and only free glucose of about 1% is S, P, O2, and H2 plus sodium (Na), potassium (K), calcium (Ca), mag-
available. Most of the glucose is converted into oligosaccharides, nesium (Mg), iron (Fe), chlorine (Cl), and silicon (Si) [25]. The leftover
polysaccharides etc. When compared with other pathways the PPP elements come about essentially in the form of trace elements for the
pathway has a higher flux than compared with the other [22]. reason that they are necessary for minor quantities. However, these
elements also included into the organic matter and are ultimately used
2.2. Nitrogen Metabolism but they are done on a dissimilar time scale [29].
Nitrogen is one of the most plentiful sources available in nature next

to C, hydrogen (H2), and O2 and is also the main contributor to the dry 4. Factors involved in the growth of algae
weight of algae. The metabolism of C and N are interconnected in mi-
croalgae [21]. The ammonium (NH4+) which is available freely is 4.1. Cultivation parameters
combined with the inorganic form, which then forms amino acids.
These amino acids require a C skeleton which can form keto acids and There are several factors which are required to measure the culti-
the energy in the form of ATP molecules will be released, which is vation of algal biomass. Some of those factors include light (based upon
necessary for the synthesis of the various amino acids like glutamate, intensity), C source and nutrient sources such as nitrates, phosphates,
aspartate, glutamine [21]. The metabolism of nitrogen sources is cat- carbohydrates and other trace elements like manganese, cobalt, zinc,
alyzed by an enzyme known as glutamine synthase. This is considered molybdenum etc. [29]. The other parameters involved are the optimal
to have a high affinity towards ammonia and can easily incorporate it temperature, optimal pH, fine mixing in the photo-reactor, removal of
into the cells [23]. O2 and uptake of CO2 in equal proportion [30]. The light, temperature,
N, and P have a close association with growth rate and lipid content of
2.3. Urea metabolism the microalgae [31]. Hence these parameters should be maintained and
controlled to be effective in reproducing the desired set of results.
Some of the algal species utilize only the nitrogen sources which are
available. Before it assimilates into the cells it is first converted into
ammonia (NH3) and bicarbonate. The two major enzymes which can 4.2. Temperature
utilize urea are urease and urea amidolyase [5]. Most of the species lack
this urease enzyme and they metabolize urea by an enzyme known as Temperature is also considered as a significant factor as well as a
UALase. The metabolic pathway in which UALase is followed is that the problematic parameter to optimize in large-scale outdoor culture sys-
allophanate lyase catalyzes the hydrolysis process of allophanate, tems such as the photo-bioreactor systems and the open pond cultiva-
which results in hydrolysis process of urea to NH3 and then to bi- tion system. Daily variations in the temperature can lead to significant
carbonate [5]. During the metabolomics analyses, the results indicate decrease in the algal lipid efficiency [29]. These algae also show a
that intermediate products in the ornithine-urea (O-U) cycle are mainly decrease in cell volume with an increase in temperature. The optimal
exhausted. With the help of the O-U cycle intermediates, a direct re- growth temperatures generally vary in between 20ºC to 30ºC [32,33].
lation can be made between both the tricarboxylic acid and the gluta- When the light intensity changes the medium and high temperatures
mine/glutamate synthase cycles. The O-U cycle consequently signifies are an environmental aspect which ultimately affects the growth of
an important metabolic pathway for anaplerotic C fixation into ni- microalgae [34]. Numerous algal species can withstand the tempera-
trogenous compounds which play an important role in the algae growth tures up to 15ºC lesser than their best, with decreased growth rates, the
[24]. temperatures higher than few degrees can lead to the death of an or-
ganism [33]. However, low evening and low seasonal temperatures
3. Biogeochemical role of algae significantly reduce the biomass productivity [35].
The acceptance choice of temperature varies with species. In the
At present, the working of the biogeochemical cycle is not totally case of freshwater microalgae, for instance, Scenedesmus and Chlorella,
misplaced. They are able to stay detained in one place for an extended are capable of adapting to the temperatures in the range of 5–35°C
period, and this habitation is known as a reservoir [25]. The study of (ideal temperature range is 25–30°C), which must be brought back into
chemical interactions between the atmosphere, hydrosphere (aquatic an ideal temperature range in the course of mass cultivation [31]. If the
systems), lithosphere (crustal minerals), and biosphere (living organ- temperature is not maintained optimally, the biochemical pathways
isms) is called Biogeochemistry [26]. In the plants and animals, the C is inside the cells may lead to damage and there will be no proper accu-
detained for a moderately small period of time in comparison with the mulation of lipids inside the cells [36]. According to study conducted
coal deposits [27]. Among the various microorganisms, algae have by Singh et al [37] some species such as Chlorella, Nannochloropsis,
played a significant function in Earth’s biogeochemistry for billions of Neochloris, Scenedesmus, Spirogyra, Chlamydomonas, Botrycoccus, Hae-
years and continue to do so today (Fig. 3). Among all the known algal matococcus, Ulva species few red algae, brown algae and blue-green
species the cyanobacteria are considered as one of the ancient organism algae can grow in a temperature range of 20ºC-30ºC with the light in-
living on Earth's crust and it generates the required amount of oxygen in tensity in the range of 33–400 mmol/m2/s [38].
the Earth. They were also responsible for generating the fossil fuels by
52
Fig. 3. Relation between algae and other organisms which are benefitted to the environmental process.
4.3. Salinity, nutrients, and pH The metabolic processes of microalgae will be significantly influenced
by the phosphate, hydrogen phosphate. Under nutrient-rich conditions,
The requirements of various factors like salinity, nutrients, and pH the mixotrophic Chlorophyceae members will show higher growth rate.
are always dependent upon the type of organisms selected. For micro- On the whole, algae require very tiny quantity of P which is available in
algal growth, the chief nutritional necessities are N and P. Certain the system and can result in ~ 30% of P which is remaining as a residue
diatoms require Si [39]. Salinity also affects the growth of algae. They in the culture [43]. In broad, algal growth has an undesirable associa-
have their own systems in adjusting the salinity array. In general, tion with lipid accumulation. Jacob-Lopes et al suggested that to avoid
seawater microalgae are capable of tolerating higher salinity conditions this concern, N famine cultivation state, and two-step cultivation has to
when compared to the freshwater microalgae. Some studies have shown be executed [44]. Prior to the experiment, one should be carefully
that algae need optimal salinity for growth. For instance, when the aware of the various growth parameters which play a significant role in
culture is provided with low salinity growth conditions, the situation the accumulation of lipids which is the prime factor for increasing the
will be supportive for the growth of algal by the addition of sodium lipid productivity. The optimum conditions which were considered
chloride (NaCl) and sodium sulphate (NaSO4). However, high salinity during the experimental procedure that favored lipid accumulation, as
(> 6 g/L) will show the adverse effect and also inhibits the growth rate well as the yield from different species, are given in Table 2.
of microalgae [40].
The pH plays a major role in the growth of algae. Under alkaline
conditions, microalgae will easily capture the CO2 from the atmosphere 4.4. Nutritional mode
and yield additional biomass [41]. The pH gradually increases to basic
as the algal growth ensues and an instantaneous increase in photo- The most common mode of nutrition for many algal species is the
synthesis and aggregation of OH- ions occurs [42]. Under acidic pH sunlight, CO2 from the environment and glucose from the nutrient
conditions (when the pH is < 5), the mainstream of the dissolved in- source. Organisms of this kind are called photoautotrophs [26]. Some
organic carbon (DIC) is CO2. On the other hand, change in pH can also species of algae can utilize pure carbon (glucose) for their growth and
impact the penetrability of the algae cell and the hydronium forms of are called the heterotrophs or the mixotrophic. The main benefit of
the inorganic salt, and continuously effect the amalgamation of the employing an organic C as the feed is that it reduces the reliance on the
inorganic salts [31]. light provision, allowing growth of conservative fermenters in the dark.
For the growth of algae, nutrients are very important such as C, O2, The most favourable growth factors should be maintained, to reach
H2, N, K, Mg, Ca, Fe, S, P, and trace minerals. The key nutrients are C, higher cell concentrations as well as to increase the volumetric pro-
O2, H2, N, P, and K. The initial three, namely C, O2, and H2 are obtained ductivity. The biomass and lipid productivities have been increased in
from water and air and the last three, namely N, P, K have to be taken the case of heterotrophic organisms when compared with the auto-
from the culture medium [29]. Throughout the farming, N, and P turn trophic [45]. Under mixotrophic circumstances, cell number will be
into the restrictive factors. They together participate in governing the amplified very quickly [46]. CO2 is also one of the controlling factors
lipid production and growth rate of microalgae. The growth, re- and the reactant factor in the photosynthesis of microalgae and plants.
production and further functional events of microalgae are strongly Increasing CO2 levels will improve the photosynthetic efficacy which
influenced by the N, which is one among the essential element. The P is leads to higher biomass yield.
one more necessary constituent aimed at the farming of microalgae.
53
Table 2
Different values of various growth parameters important and required for Algae.
Strain Habitat Nutrients Biomass Lipid Ref.
Biomass yield Lipid production Total lipid

production (g/L) (g/L/day) extracted
(g/L/day) (wt% of biomass
P. carterae Marine water Modified f/2 medium 0.22 n.a 0.072 n.a [129]
D. salina Marine water j/2 medium n.a n.a 0.116 n.a [129]
Porphyridium cruentum Marine water n.a 0.37 n.a 0.034 9.5 [130]
Tetraselmis suecica (F&M-M33) Marine water n.a 0.32 n.a 0.027 8.5 [130]
Tetraselmis sp. (F&M-M34) Marine water n.a 0.3 n.a 0.043 14.7 [130]
Tetraselmis. Suecica (F&M-M35) Marine water n.a 0.28 n.a 0.036 12.9 [130]
Phaeodactylum tricornutum (F&M-M40) Marine water n.a 0.24 n.a 0.044 18.7 [130]
Nannochloropsis sp. (F&M-M26) Marine water n.a 0.21 n.a 0.061 29.6 [130]
Ellipsoidion sp. (F&M-M31) Marine water n.a 0.17 n.a 0.047 27.4 [130]
Nannochloropsis (CS 246) Marine water n.a 0.17 n.a 0.049 29.2 [130]
Isochrysis sp.(CS 177) Marine water n.a 0.17 n.a 0.037 22.4 [130]
Pavlova salina (CS 49) Marine water n.a 0.16 n.a 0.049 30.9 [130]
Pavlova lutheri (CS 182) Marine water n.a 0.14 n.a 0.052 35.5 [130]
Isochrysis sp. (F&M-M37) Marine water n.a 0.14 n.a 0.037 27.4 [130]
Skeletonema sp. (CS 252) Marine water n.a 0.09 n.a 0.027 31.8 [130]
Thalassiosira pseudonana (CS 173) Marine water n.a 0.08 n.a 0.017 20.6 [130]
Skeletonema costatum (CS 181) Marine water n.a 0.08 n.a 0.017 21.1 [130]
Chaetoceros muelleri (F&M-M43) Marine water – 0.07 n.a 0.021 33.6 [130]
Chaetoceros calcitrans (CS 178) Marine water n.a 0.04 n.a 0.017 39.8 [130]
Chlorococcum sp. (UMACC 112) Fresh water n.a 0.28 n.a 0.053 19.3 [130]
Scenedesmus sp. Fresh water n.a 0.26 n.a 0.053 21.1 [130]
Chlorella sorokiniana (IAM-212) Fresh water n.a 0.23 n.a 0.044 19.3 [130]
Chlorella sp. (F&M-M48) Fresh water n.a 0.23 n.a 0.042 18.7 [130]
Scenedesmus sp. (F&M-M19) Fresh water n.a 0.21 n.a 0.04 19.6 [130]
Chlorella vulgaris (F&M-M49) Fresh water n.a 0.20 n.a 0.369 18.4 [130]
Scenedesmus quadricauda Fresh water n.a 0.19 n.a 0.351 18.4 [130]
Monodus subterraneus (UTEX 151) Fresh water n.a 0.19 n.a 0.03 16.1 [130]
Chlorella vulgaris (CCAP 211/11b) Fresh water n.a 0.17 n.a 0.032 19.2 [130]
Porphyridium cruentum Marine water n.a 1.5 1.90 n.a n.a [131]
Chlorella vulgaris Fresh water BBM 0.020 3.2 0.002 27 [132]
Aphanothece Fresh water BGN 0.031 5.0 0.005 8 [132]
Dunaliella Marine water ESAW 0.015 2.4 0.002 17.1 [132]
Phaeodactylum Marine water Ukeles 0.0003 0.150 0.000 6.1 [132]
Phormidium BGN 0.017 2.8 0.002 11.7 [132]
Scenedesmus Fresh water BBM 0.027 4.3 0.003 14.1 [132]
Neochloris oleabundans Fresh water Bristol medium 0.15 0.09 0.038 56 [133]
Chlorella sp. Marine water Walne’s nutrient medium n.a 1.42 0.139 26 [134]
Chlorella vulgaris Fresh water Basal medium 0.254 1.69 0.054 38 [135,136]
B. braunii Fresh water modified 0.026 n.a 0.005 25.7 [137]
chu13 medium
C. vulgaris Fresh water BG11 medium 0.104 n.a 0.006 11.9 [137]
Scenedesmus sp. Fresh water BG11 medium 0.217 n.a 0.020 11.9 [137]
Scenedesmus obliquus Fresh water N-deficient culture medium 0.09 2.0 n.a 35 [133]
Chlorella vulgaris Fresh water N-deficient culture medium 0.18 3.0 n.a 40 [133]
Neochloris oleoabundans Fresh water N-deficient culture medium, 0.09 2.1 n.a 35 [133]
Spirulina maxima Fresh water N-deficient culture nedium 0.21 3.1 n.a 9 [133]
N. oculata (NCTU-3) Marine water modified 0.48 n.a 0.142 29.7 [138]
f/2 medium
N. oleoabundans Fresh water Bristol medium 0.19 1.96 0.004 16.5 [5]
S. obliquus Fresh water Bristol medium 0.26 1.87 0.03 12.5 [5]
Chaetoceros muelleri (F&M-M43) Marine water n.a 0.07 n.a 0.021 33.6 [139]
Chaetoceros calcitrans (CS 178) Marine water n.a 0.04 n.a 0.017 39.8 [139]
P. tricornutum (F&M-M 40) Marine water n.a 0.24 n.a 0.044 18.7 [139]
Skeletonomacostatum (CS 181) Marine water n.a 0.08 n.a 0.017 21 [139]
Skeletonoma sp.(CS 252) Marine water n.a 0.09 n.a 0.027 31.8 [139]
Thalassioria pseudonana (CS 173) Marine water n.a 0.08 n.a 0.017 20.6 [139]
C. vulgaris (F&M-M49) Fresh water n.a 0.20 n.a 0.036 18.4 [139]
Scenedemus quadricauda n.a n.a 0.19 n.a 0.035 18.4 [139]
Scenedemus (F&M-M19) Fresh water n.a 0.21 n.a 0.040 19.6 [139]
Scenedemus sp. DM Fresh water n.a 0.26 n.a 0.053 21.1 [139]
T. suecica (F&M-M33) Marine water n.a 0.32 n.a 0.027 8.5 [139]
(continued on next page)
54
Table 2 (continued)

Monodus subterraneus (UTEX 151) Freshwater n.a 0.19 n.a 0.030 16.1 [139]
Nannochloropsis sp. (CS 246) Marine water n.a 0.17 n.a 0.049 29.2 [139]
Isochrysissp. ((T-ISO) CS 177) Marine water n.a 0.17 n.a 0.037 22.4 [139]
Isochrysissp. (F&M-M37) Marine water n.a 0.14 n.a 0.037 27.4 [139]
Chaetoceros muelleri Marine water n.a 0.07 n.a 0.021 33.6 [30]
Chaetoceros calcitrans Marine water n.a 0.04 n.a 0.017 16.4 [30]
Chlorella emersonii Fresh water n.a 0.041 n.a 0.050 25 [30]
Chlorella protothecoides Fresh water n.a 7 n.a 1.24 57 [30]
Chlorella sorokiniana Fresh water n.a 1.47 n.a 0.044 22 [30]
Chlorella vulgaris Fresh water n.a 0.20 n.a 0.04 58 [30]
Chlorella sp. Fresh water n.a 2.5 n.a 0.042 48 [30]
Chlorococcum sp. Fresh water n.a 0.28 n.a 0.53 19.3 [30]
Dunaliella salina Marine water n.a 0.34 n.a 0.116 25 [30]
Ellipsoidion sp. n.a n.a 0.17 n.a 0.047 27.4 [30]
Isochrysis sp. Marine water n.a 0.17 n.a 0.037 33 [30]
Monodus subterraneus Fresh water n.a 0.19 n.a 0.030 16 [30]
Nannochloris sp. Marine/fresh n.a 0.51 n.a 0.076 56 [30]
Water/brackish
Nannochloropsis oculata Brackish water n.a 0.48 n.a 0.142 29.7 [30]
Nannochloropsis sp. Marine/fresh n.a 1.43 n.a 0.090 53 [30]
Water/brackish
Pavlova salina Marine water n.a 0.16 n.a 0.049 30.9 [30]
Pavlova lutheri Marine water n.a 0.14 n.a 0.040 35.5 [30]
Phaeodactylum tricornutum Marine water n.a 1.9 n.a 0.044 57 [30]
Porphyridium cruentum Marine water n.a 1.50 n.a 0.034 60 [30]
Scenedesmus sp. Fresh water n.a 0.26 n.a 0.053 21.1 [30]
Skeletonema sp Marine water n.a 0.09 n.a 0.027 31.8 [30]
Skeletonema costatum Marine water n.a 0.08 n.a 0.017 51.3 [30]
Thalassiosira pseudonana Marine water n.a 0.08 n.a 0.017 20.6 [30]
Tetraselmis suecica Marine water n.a 0.32 n.a 0.036 23 [30]
Tetraselmis sp. Marine water n.a 0.30 n.a 0.043 14.7 [30]
Scenedesmus sp. Fresh water BG 11 0.217 0.003 0.002 n.a [138]
Botryococcus braunii Fresh water Modified chu13 0.026 n.a 0.005 0.005 [138]
Chlorella vulgaris Fresh water BG 11 0.104 n.a n.a 0.020 [138]
Botryococcus sp. Fresh water n.a 0.035 n.a 0.011 n.a [140]
Chlorella vulgaris Fresh water n.a 0.074 n.a 0.011 n.a [140]
Scenedesmus sp Fresh water n.a 0.071 n.a 0.009 n.a [140]
Scenedesmus sp. Fresh water 50% BG 11 0.11 n.a 0.008 31–33 [141]
Botryococcus braunii Fresh water BG 11 n.a 0.037 n.a 13.5 [142]
Chlorella saccharophila Fresh water BG 11 n.a 0.002 n.a 18.10 [142]
Dunaliella tertiolecta Marine water Modified BG 11 n.a 0.038 n.a 15.20 [142]
Pleurochrysis carterae Marine water Modified BG 11 n.a 0.037 n.a 12 [142]
Consortium Fresh water BG 11 n.a 0.041 n.a 12.20 [142]
Chlorella Fresh water Artificial wastewater n.a 0.69 0.147 42 [143]
Vulgaris medium
Chlorella sp Fresh water Tris–acetate–phosphorus 0.92 1.07 0.200 n.a [41]
Botryococcus braunii Fresh water Secondary domestic 0.034 0.48 n.a 36.14 [13]
wastewater
T. suecica Marine water f/2 media 0.064 0.58 n.a n.a [38]
P. tricornutum Marine water f/2 media 0.018 0.26 n.a n.a [38]
C. calcitrans Marine water f/2 media 0.044 0.48 n.a n.a [38]
I. galbana Marine water f/2 media 0.024 0.57 n.a n.a [38]
N. oculata Marine water f/2 media 0.020 0.57 n.a n.a [38]
Chaetoceros muelleri (F&M-M43) Marine water n.a 0.07 n.a 0.021 33.6 [144]
Chaetoceros calcitrans Marine water n.a 0.04 n.a 0.017 39.8 [144]
P. tricornutum (F&M-M40) Marine water n.a 0.24 n.a 0.044 18.7 [144]
Skeletonema costatum (CS 181) Marine water n.a 0.08 n.a 0.017 21.0 [144]
Skeletonema sp. (CS 252) Marine water n.a 0.09 n.a 0.027 31.8 [144]
Thalassiosira pseudonana (CS 173) Marine water n.a 0.08 n.a 0.017 20.6 [144]
55
Table 2 (continued)

C. vulgaris (F&M-M49) Fresh water n.a 0.20 n.a 0.036 18.4 [144]
Scenedesmus (F&M-M19) Fresh water n.a 0.21 n.a 0.040 19.6 [144]
Scenedesmus sp. DM Fresh water n.a 0.26 n.a 0.053 21.1 [144]
Tetraselmis suecica (F&M-M33) Marine water n.a 0.32 n.a 0.027 8.5 [144]
Monodus subterraneus Fresh water n.a 0.19 n.a 0.030 16.1 [144]
Nannochloropsis sp. (CS 246) Marine water n.a 0.17 n.a 0.049 29.2 [144]
Nannochloropsis sp. (F&M-M280 Marine water n.a 0.17 n.a 0.060 35.7 [144]
Isochrysis sp. (T-ISO) CS 177) Marine water n.a 0.17 n.a 0.037 22.4 [144]
Isochrysis sp. (F&M-M37) Marine water n.a 0.14 n.a 0.037 27.4 [144]
Scenedesmus sp. (LX1) Fresh water BG11 medium n.a 313a 112b n.a [141]
Chlorella emersonii Terrestrial M7 medium 3.7 37.4 n.a n.a [145]
Botrycoccus braunii Fresh water M7 medium 4.6 36.1 n.a n.a [145]
S. obliquus (YSL02) Fresh water Bold basal medium 1.84 n.a 0.53 29 [146]
Chla. Pitschmannii (YSL03) Fresh water Bold basal medium 1.04 n.a 0.54 51 [146]
C. vulgaris (YSL04) Fresh water Bold basal medium 1.65 n.a 0.44 26 [146]
S. obliquus (YSL05) Fresh water Bold basal medium 1.71 n.a 0.48 28 [146]
Chla. Mexicana (YSL07) Fresh water Bold basal medium 1.53 n.a 0.45 29 [146]
C. vulgaris (2714) Fresh water Modified culture medium 0.39 1.17 0.16 40 [147]
Chlorella vulgaris Fresh water N11 medium n.a 0.31 0.171 55 [148]
Chlamydomonas reinhardtii Fresh water n.a 2.0 n.a 0.505 25.25 [113]
Scenedesmus obliquus Fresh water n.a 0.026 n.a 0.008 31.14 [113]
Botryococcus braunii Fresh water n.a 0.345 n.a 0.062 17 [113]
Chlorella vulgaris Fresh water Kessler and czygan 0.16 n.a 0.034 30 [149]
Scenedesmus obliquus Fresh water Kessler and czygan 0.25 n.a 0.041 60 [149]
Ellipsoidion Parvum Fresh water Kessler and czygan 0.09 n.a 0.111 n.a [149]
C. oleofaciens n.a BG11 medium 0.20 n.a 0.035 n.a [149]
H. pluvialis Fresh water BG11 medium 0.06 n.a 0.020 n.a [149]
B. braunii Fresh water BG11 medium 0.03 n.a 0.43 40 [149]
Scenedesmussp Fresh water TAP media 0.080 n.a 0.030 n.a [150]
Chlamydomonas debaryana Fresh water/ B3NV media 0.051 n.a 0.005 n.a [150]
Terrestrial
Chlorella sorokiniana-(FGP5) Fresh water n.a 0.030 n.a 0.003 n.a [150]
Nannochloropsis sp Marine/ n.a 0.21 n.a 0.061 29.6 [151]
Freshwater
Chlorella vulgaris Fresh water Thin stillage (TS) 2.5 9.8 1.1 43 [152]
Chlorella vulgaris Fresh water Soy whey (SW) 1.6 6.3 0.2 11 [152]
Chlorella vulgaris Fresh water Modified basal medium 2.0 8.0 0.6 27 [152]
B. braunii (AP103) Fresh water Modified 0.114 1.8 n.a 19 [153]
chu13 medium
T. variabilis Brackish water BG110 0.040 0.020 n.a 12.1 [154]
A. augstumalis n.a BG110 0.017 0.008 n.a 3.2 [154]
P. autumnale n.a Modified BG11 0.055 0.027 n.a 2.4 [154]
Nannochloropsis Oculata Marine water n.a 0.004 0.002 n.a 15.1 [154]
Spirulina sp Fresh water Zarrouk’s medium 1.37 n.a 0.66 20 [155]
Chlorella sp. Fresh water BBM medium 1.65 n.a 0.74 26 [155]
Amphora sp. Marine water n.a 0.16 n.a 0.037 24 [156]
Chlorella vulgaris Fresh water n.a 0.46 n.a 0.079 17.3 [156]
Chlorella salina Marine water n.a 0.17 n.a 0.0182 11 [156]
Chlorella protothecoides Terrestrial n.a 0.25 n.a 0.045 18 [156]
Chlorella emersonii Terrestrial n.a 0.29 n.a 0.054 18.6 [156]
Scenedesmus sp. Fresh water n.a 0.10 n.a 0.015 16 [156]
Ankistrodesmus sp. n.a n.a 0.09 n.a 0.015 17.5 [156]
Chlamydomonas reinhardtii Fresh water n.a 0.05 n.a 0.009 18.9 [156]
D. salina (Shariati) Marine water n.a 0.05 n.a 0.010 18.9 [156]
Dunaliellasp Marine water n.a 0.12 n.a 0.025 22 [156]
D. salina (UTEX 200) Marine water n.a 0.15 n.a 0.036 24 [156]
Chlorella pyrenoidosa Fresh water Bold’s basal medium 0.106 n.a 0.019 29.68 [157]
56
Table 2 (continued)

Isochrysis galbana Marine Enriched artificial 0.51 1.51 0.070 16.47 [158]
seawater with f/2 medium
Kirchneriella lunaris Fresh water BG11 medium 0.293 n.a 0.008 n.a [159]
Selenastrum capricornutum Fresh water BG11 medium 0.097 n.a 0.006 n.a [159]
Staursatrum sp. Fresh water BG11 medium 0.078 n.a 0.000 n.a [159]
Chlorella vulgaris Fresh water BG11 medium 0.225 n.a 0.007 n.a [159]
Scenedesmus obliqnus Fresh water BG11 medium 0.206 n.a 0.006 n.a [159]
Navicula sp. Fresh water D1 medium 0.071 n.a 0.003 n.a [159]
Phaeodactylum tricornutum Fresh water f/2 medium 0.256 n.a 0.026 61.43 [159]
Batrachospermum Sirodotia Fresh water BG11 medium 0.049 n.a 0.001 n.a [159]
Lyngbya kuetzingii Fresh water BG11 medium 0.234 n.a 0.007 n.a [159]
Isochrysis sphacrica Fresh water f/2 medium 0.255 n.a 0.008 n.a [159]
Microcystis aeruginosa (NPCD-1) Fresh water n.a 0.046 13.1 0.013 28 [160]
Synechococcus sp. (PCC7942) Marine water n.a 0.052 n.a 0.014 26.9 [160]
Trichormus sp. (CENA77) Soil, subglacial n.a 0.030 n.a 0.007 23.7 [160]
Chlorella protothecoides Terrestrial Bristol’s and Wu’s Media 8.7 43.3 4.32 42.3 [161]
culture medium
Ettlia sp. (YC001) Fresh water BG11 Agar medium 0.19 3.10 0.080 42 [162]
Aurantiochytrium sp. (KRS101) Marine water Defined medium 6.69 31.8 n.a 38.1 [163]
Chlorella protothecoides Terrestrial Basal culture medium 0.5 0.015 27 [164]
Endogenous Chlorella sp n.a Brewery wastewater n.a 2.7 0.052 23 [14]
Chlorella vulgaris (UTEX-265) Fresh water TAP medium n.a 3.5 0.108 42 [14]
Ettliatexensis Fresh water Bold’s basal medium 0.92 0.459 0.322 35 [15]
Synechococcus sp. (PCC7942) Marine water BG-11 liquid medium 0.124 n.a 0.35 29 [165]
Chlorella sp. (KMN1) Marine water Bold’s basal medium 0.022 0.96 0.26 27.11 [166]
Scenedesmus sp. (KMN4) Marine water Bold’s basal medium 0.023 0.92 0.25 28.63 [166]
Monoraphidium sp. (KMN5) Marine water Bold’s basal medium 0.013 0.65 0.23 34.93 [166]
Chlorococcum sp. (IMMTCC-1) Fresh water Bold basal medium n.a n.a 2.095 8.7 [167]
Chlorella sp. (IMMTCC-2) Fresh water Bold basal medium n.a n.a 4.071 22.7 [167]
Scenedesmus sp. (IMMTCC-3) Fresh water Bold basal medium n.a n.a 2.429 11.04 [167]
Scenedesmus sp. (IMMTCC-7) Fresh water Bold basal medium n.a n.a 2.786 14.6 [167]
Micractinium sp. (ME05) Fresh water BG-11 medium 0.47 0.93 0.05 10.7 [168]
H. tetrachotoma (ME03) n.a BG-11 medium 0.04 0.31 0.01 8.7 [168]
Scenedesmus sp. (ME02) Fresh water BG-11 medium 0.06 0.12 0.004 12.3 [168]
Ettlia sp. Fresh water Sugar factory wastewater n.a 8.02 0.96 42 [46]
I. galbana Marine water f/2 medium n.a 0.001 0.013 21 [36]
P. tricornutum Marine water f/2 medium n.a 0.001 0.017 28 [36]
Note: n.a – not available.

a
– per g biomass.
b
– per g lipid.
5. Techniques involved in the processing of algae higher yield of biomass, with less operating cost [50]. However, while
designing an efficient harvesting technique a few points should be given
5.1. Harvesting techniques importance.
Harvesting is defined as a sequence of process for eliminating water • The choice of microalgae and the desired products
content from the growth culture of algae with the help of various • A complete cell separation process for efficient recycling, that con-
downstream techniques available [47]. It can be also defined as diluting tributes to the low cost of down streaming processing.
the concentrated microalgal culture or suspension to slurry or paste. • The chosen technique should have minimal impact on the further
Keeping in mind the cost of extraction, the downstream processes must processes [51].
be reduced for an efficient extraction process [48]. The basic process for
harvesting is the collection of individual algae cells or the medium upon The harvesting process involves two steps
which they are grown which are analyzed [23]. Generally, the most
widespread harvesting processes include screening, coagulation, floc- a) Bulk harvesting: A bulk suspension of biomass undergoes sedi-
culation, flotation, sedimentation, filtration, and centrifugation [23]. mentation.
On the other hand, there are also other techniques like electrophoresis, b) Thickening: This process is done to separate the biomass and to
electroflotation, and ultrasound which are of less importance [49]. concentrate the slurry matter with the help of filtration and cen-
Hence the selection of harvesting procedure should be based on energy trifugation.
efficiency and cost factor.
An efficient harvesting technique should take into consideration Many of the current harvesting techniques have numerous draw-
various parameters of algae like size and density, in order to achieve a backs which have an impact on the cost and quality of the products.
57
Harvesting is a challenging task when biofuel production is done on a cells active. Hence microalgae can be used in the treatment of
commercial scale [52]. The reason for high cost for biofuel production wastewater for the removal of excess phosphate [59]. When the pH is
is mainly due to these harvesting techniques. They account for about high flocculation is caused due to the formation of inorganic
20–30% of the total cost of algal biomass [52]. Though there are many precipitates, thus after harvesting biomass consists of the excess
harvesting techniques, till date no specific technique has been re- amount of minerals. This process is performed by the addition of
commended for filamentous algae owing to their structural organisa- metallic salts, an alkaline compound, or polyelectrolytes. The alkaline
tion. compounds such as sodium hydroxide (NaOH), potassium hydroxide
(KOH), calcium hydroxide (Ca(OH)2), or magnesium hydroxide (Mg
5.1.1. Centrifugation (OH)2) cause biomass accumulation [60]. However, the flocculation is
Centrifugation is a method in which separation of two immiscible reliant on the microalgae cell density logarithm. It is not linearly
liquids takes place with the help of centripetal force. The particle size related to the quantity of algae biomass [61]. During the process, the
and density are two crucial factors in a centrifugation process. So much pH possibly will vary and will influence the further downstream of the
research was done on centrifugation techniques an interesting result biomass processing.
was found [34]. Heasman et al [49] stated that 90–100% harvesting The metal ions like Mg2+ and Ca2+ play an important role in
effectiveness can be attained via centrifugation. Sim et al [53] reported flocculation process after increasing the pH [58]. During the growth of
that centrifugation is the most efficient technique but highly costly the species in the medium, it was found that the negatively charged
when used for production on a commercial scale. Generally, this type of bodies were hydrolysed into positive precipitates by the process of
technique is applied for the production of secondary metabolites [52]. sweeping flocculation. In contrast to an important role in flocculation
When the algal culture is separated by centrifugation, high gravita- process by increasing pH, there is also a possibility that the ions have
tional force and shear stresses are applied in the process that might played a crucial part in decreasing pH [62].
damage the cell structure. The mechanism of flocculation is dependent upon the physico-
The various types of centrifugation techniques or systems available chemical properties of microalgae cells. Since the surface of the mi-
are as follows: croalgae cells is negatively charged, the zeta potential of microalgae is
explored during the flocculation process [58]. The zeta potentials show
• Hydro-cyclone a quick rise from a range of pH 6.5–4.0 and also the equivalent floc-
• Solid bowl decanter culation efficacies also increase to the highest with the drop in pH [60].
• Nozzle type When the pH is greater than 6.0, the surface charge of the microalgae
• Solid ejecting disc cells is subjected to the neutral amine groups (R-NH2) and the nega-
tively charged ions (carboxylate) [60]. The carboxylate ions would
5.1.2. Flocculation accept the protons (H+), at that moment the surface charge is reduced
It is a method in which scattered units are collected together to form and the algae cells become unstable and are clotted to form big flocks
huge units and made to settle down. This method is an extensively used [55]. The flocculation mechanisms are at the maximum when the sur-
technique in diverse activities ranging from brewing to wastewater face charge of the algae cells is completely neutralized. As soon as the
treatment and mining etc. [54]. In recent years’ various flocculation pH values drop from 4.0 to 1.5, the zeta potential constantly increases
techniques have been explored ranging from chemical flocculation to with equivalent flocculation [60]. In the exponential growth phase, the
the latest bio-flocculation. In chemical flocculation process, several biomass is high whereas in lag phase it remains low [62]. In stationary
multivalent metal particles like ferric chloride (FeCl3), ferric sulphate phase as well as the exponential phase the cells form clumps and
(Fe2(SO4)3), aluminium chloride and aluminium sulphate which is clusters. Since the surfaces of the cells are neutralized the heavier cells
commonly known as alums are present which interfere with the har- settle easily when compared to the single cells. Hence the flocculation
vesting procedures [23,55]. During the process, these particles stay on efficiencies are greater with the rise in biomass concentration [62].
the surface of the biomass and interfere with the extraction of lipids. Some of the prominent flocculants and their optimum pH are listed in
When compared with the synthetic flocculants, natural flocculants are Table 3. The Flocculation process varies with different inorganic and
safer to act together with the negative surface of the cells [56]. A good organic salts and is classified based on their chemical composition.
example for flocculant is chitosan which is very effective but works only
at very acidic pH. Therefore, cationic starch which is independent of pH
and charge is considered as a substitute for chitosan [57]. 5.1.2.3. Inorganic flocculation. As we know that the cells of the
microalgae are negatively charged and the ions in the chemicals
5.1.2.1. Bioflocculation. This process occurs in the lakes or ponds interact with these, hence disrupting the algae cells resulting in
spontaneously because of the extracellular polymer substance. successful harvesting [60]. The flocculation of the microalgae occurs
However, this particular mechanism is poorly understood and a lot of at a considerably low pH. The flocculants with high charge density are
extensive research needs to be carried out [58]. This method is cost and considered to be the best flocculants. Among this Alum is the best
energy efficient alternative harvesting method. This is usually used in flocculant in operation during the wastewater treatment, but the only
the process of treating wastewaters [55]. Flocculation can also be disadvantage is that it may hinder the impurities during the lipid
influenced by the rapid increase in the pH, temperature or nutrient extraction stage [55,56]. To avoid this problem any negatively charged
depletion, and changes in dissolved oxygen. Generally, these techniques
Table 3
are not used for pre-harvesting. When this process of flocculation is
Some of the prominent flocculants and their optimum pH.
carried along with the mixed bacterial source then an extra energy
should be invested in supplying nutrients otherwise there could be S. No Flocculant Type of ion Optimal pH Water References
contamination [56,58]. system
1. Alum Polyvalent 5.3–5.6 Wastewater [169]

5.1.2.2. Auto flocculation. This process automatically occurs at basic Metal Ion system
pH due to CO2 depletion. The major precipitates which are formed in 2. Lime Positively 10.5–11.5 Wastewater [170]
the auto-flocculation process are Ca and Mg precipitates [58]. Since Treatment charged systems [169]
Precipitation metal
these Ca surfaces are positively charged they can interact easily with
hydroxide
the negatively charged surfaces of the algal cells which results in the precipitates
reserves for the phosphate sources and making the surfaces of the algal
58
surfaces such as Iron or Aluminium are added to the surface of the • Height
microalgal cells to neutralise the medium [63]. • Time of settling
• The specific gravity suspension and
5.1.2.4. Organic flocculation. The organic flocculants are otherwise • The specific gravity of particles [69].
known as polyelectrolytes. The strength of these polymers is
dependent on certain properties like the charge of the cells, pH, the 5.1.4. Filtration
concentration of biomass. The high concentration of biomass will help It is a mechanical or physical process which is used for separating
in the flocculation process. Mixing of the biomass at low concentrations solids from liquids or gasses by interposing a medium through which
brings the cells together, but if the shear forces are high it can disrupt fluid can only pass [23]. For this process filters of particular pore size
the flocs [63]. Apart from these the functional charges on the algal cell called micro, strainers are used [54,70]. These are used as rotating
walls play an important role in the formation of the negative charge filters with frequent backwash. This filtration technique is usually used
centers on the cell surface. The size of the floc in an organic in filtering largely sized algae such as filamentous species. This process
sedimentation should be greater than 100 µm for efficient is not appropriate for microalgae like Chlorella, Dunaliella, Scenedesmus
flocculation. Pushparaj et al [64] have shown that the aluminium etc. Several kinds of filtration processes, such as vacuum filtration,
sulphate (Al2(SO4)3) was effective, meant for successful flocculation for micro-filtration, ultra-filtration, dead-end filtration, pressure filtration,
algal harvesting. The most successful flocculants for the revival of algal and tangential flow filtration (TFF) [71]. According to the recent stu-
species are the cationic flocculants since they would be attracted to the dies, it is shown that TFF and pressure filtration can be used effectively
cells of algae which are negatively charged. Chitosan is considered as since they consume less energy. The major drawback of this filtration is
the best biodegradable bioflocculant which is used in the water the frequent change of filters and membranes which makes the process
purification process. Since this Chitosan is too expensive, the economically not viable [70].
economically feasible polymeric flocculants are generally used in
flocculating the algae species [63]. 5.1.5. Microstrainers
Microstrainers are widely used to separate algae from water streams
of reservoirs before treating the water. Typically, a centrifugal micro
5.1.2.5. Combined flocculation. The combined flocculation process
strainer consists of a sealed tubular container with a tubular screen
involves the combination of two or more different flocculants.
attached, with together revolving about an empty tube making a shared
Sukenik et al [65] studied that marine algae have found two existing
central axis [50]. The solid-liquid mixture is passed through the tube
different methods for inducing two different flocculation methods.
and is operated centrifugally. The mesh size range is 15–64 µm which
Different low concentrations of chitosans in combination with
allow only very fine particles. The suspended particles which are very
inorganic flocculants like (FeCl3, Alum etc.) are used. During the
large in size cannot pass through, remain on its external side.
experiment, flocculant dosages were reduced. The Ferric chloride
The major advantages of these micro strainers micro strainers are:
dosage was reduced from 100 mg/L to 50 mg/L without/with
• The simplicity of manufacturing and functioning

addition (2.5 mg/L) of chitosan. This was observed in species of
• Ease of operation.
Isochrysis galbana. In the same way for C. stigmatophora, the
• Low investment.
concentration was also reduced to 50% upon addition of FeCl3.
Chitosan played a key role in decreasing the algal flocculation. When
• Low energy consumption
• High filtration ratios.
Chitosan in combination with FeCl3 did not improve algal removal. The
reason for this is that Chitosan molecules fail to bridge between these
particles at a high ionic strength which reduces the charge, which also
The major problems encountered while using these micro strainers
reduces the effectiveness dose [65].
are:
5.1.2.6. Electrolytic process. In this process, the movement of

• Solids are not removed completely.
microalgae is subject to the movement of charges from the anode to
• Handling of solids is difficult during fluctuations.
cathode and they form aggregates [66,67]. This process basically
involves three different mechanisms:
• Algae and bacterial films forms on the surface of the micro fabric.
During the filtration process if the concentration of algae is very
• Coagulants production by the electrolytic oxidation of the sacrificial high then these concentrated algae can block the screen whereas the
electrode. smaller algae can be run off through the screen very easily [50]. Pet-
• Deterioration of particulate suspension and breaking of the emul- ruvski et al [71] have recovered a maximum of 70–90% of the fresh-
sion. water algae through the tangential flow filtration. The main benefit of
• Destabilized phase accumulation which can form flocs. this filtration process is that it retains the structure, motility, and
properties of the collected algae.
Based on the existing literature [66], it was shown that when there
is electrolytic flocculation applied there is an approximately 80–95% of 5.1.6. Electrophoresis process
algae is removed [67]. It is defined as the movement of charged particles from one end to
the other end under the influence of an electric field, which pushes the
5.1.3. Gravity sedimentation charged algae to shift out of the tray. This technique is another method
Technically this is a method where the heavier algal particles settle which efficiently utilizes and separates the algae without adding any
down to the bottom. Hence we can easily detect the lighter and the chemicals. The major advantages of this technique are environmental
heavier algal particles separately. The major factors which decide and compatibility, adaptability, energy effectiveness, safety, selectivity and
influence the gravity sedimentation are the radius and density of the price effectiveness [72].
algal cells. This technique is generally used in separating algae in The key factors for separation of the particles in the electrophoresis
wastewater applications. Edzwald [68] revealed the fact that the low include:
mass microalgal units do not resolve well and are not efficiently sepa-
rated by the settling process. Some of the important factors which affect • Sample
the settling of algal particles are • Electric Field
59
• Medium handling of huge quantity of samples is the key advantage of this

• Buffer method. Various other advantages and disadvantages are listed in the
Table 4.
Generally, the algal particles are negatively charged. Throughout
the process when a tray of electrophoresis consists of the growth 5.2.1.2. Bligh and Dyer method. This technique is quite similar to the
medium an electric current is passed. Pearsall et al. [73] conducted a folch method but majorly varies in the ratio of solvents to the tissues
comparable kind of experiment and proved that the effects of electro- [80]. In this method, the proteins from the sample are precipitated
phoresis are complex due to its fluid motion. During the fluid move- between the two lipid phases, both lipid extraction and partitioning are
ment, it appears that the algal cell and liquid activity can be adequately accomplished simultaneously. The combination of the solvent can also
strong which can pressurize the fluid movement or lead the perfor- change based on the polarity of the lipids present in the species of algae.
mance of the fluid. Sandbank et al. [74] have shown that extensive Apart from the main technique by the Bligh and Dyer, many researchers
variety of microalgae species were harvested by the electroflotation globally have also modified depending on the situation and the
method with 5% of solids harvested in algae. condition of the laboratories. The most common method used for
modification was done by the [81]. They have shown that they have
5.1.7. Ultra-Sonication replaced water with 1 M NaCl so that it can evade attachment of acidic-
It is defined as an act of employing energy associated with the vi- lipids to denatured-lipids [82]. From the previous extraction
bration of matter (well known as sound energy) to disturb particles in a procedures, it was also known that by adding a salt solution, the
sample. The frequency of > 20 kHz is generally used. Bosma et al. [75] yield of lipid improves with a shorter separation time. In the same way,
successfully applied this technique to demonstrate that the ultra- adding 0.5% of CH3COOH to the water-phase improved the retrieval of
sonication method is an advantageous method. In this, the efficiencies acidic-phospholipids. Among the above processes, Hajra [81] method
were higher than 90% and the flow rate varied between 4 and 6 L/day. was found to be the most efficient process which has been demonstrated
Li et al. [41] have shown that ultrasound waves could improve the till now.
removal by the coagulation of one of the toxic algae namely Microcystis Regarding various advantages and disadvantages of the above
aeruginosa. The major process involved in the destruction of algal cells methods, many results were discussed for Folch and Bligh & Dyer
in this process is the ultrasonic irradiation of gas vacuoles. The algal methods. Many of them published results, which were neither validated
cells perform as “nuclei” for acoustic cavitation and fall down in the nor described. Many researchers even indicated the modifications
course of “bubble crush” period, which resulted in the settlement. which were made. However, the research conducted by Iverson [83]
Collection and absorption of microalgae biomass cost highly to the have shown that Bligh and Dyer's procedure is more effective and re-
overall procedure of cultivation. Therefore, much more resourceful and liable. The results obtained from Bligh and Dyer had identical lipid
profitable harvesting technologies have to be developed to improve the samples when compared with the Folch method where the tissue
commercial as well as for the industrial capacity [76]. sample lipid was taken (< 2%). After many experiments conducted by
many researchers, they have concluded that both the methods em-
5.2. Lipid extraction techniques ployed gave or produced accurate results out of which some of them
have used high sample/solvent ratio, which could not alter the lipid
In recent years, microalgae are treated as the most potential and content significantly. If the samples contained a greater amount of lipid
valuable feedstock for the source of biofuels. Various products such as then in such samples an increase solvent/sample ratio is necessary for
bio-ethanol, biodiesel, bio-butanol, bio-methane (bio-CH4), jet fuel, quantitative lipid evaluation [83]. Various other advantages and dis-
biohydrogen (bio-H2), and thermo chemical transformation products advantages are listed in the Table 4.
such as bio-oil, biocrude, and syngas are feasibly produced from mi- Later many researchers have tried to modify the solvents used in the
croalgae [23]. Globally huge amount of study is being carried out in procedure of Bligh and Dyer. Considering the Chloroform solvent as a
improving the lipid yield of microalgae. This lipid extraction is con- toxic substance [84] were the first to substitute (or) change the solvent
sidered as one of the most important processes for extracting biodiesel system to hexane-isopropanol. This result showed the lower efficiency
from microalgae. In the development of algal lipids, there are many of extracted substance (lipid). Many years later after a careful ex-
techniques available for the extraction of algae. We will discuss those amination of the solvent few researchers replaced the chloroform-me-
techniques in brief with more emphasis on the cost-effective methods. thanol to heptane and ethyl acetate mixture [85]. In the point of en-
In practical, the extraction of lipids from the algal sources has been vironmental protection, these hazardous chemical combinations should
divided into two major parts like be replaced by bio solvents [77]. The basic method involves the addi-
tion of 1 M NaCl instead of water which helps in preventing the binding
a) Lipid extraction by chemicals and solvents. of acidic lipids to denatured lipids. Furthermore, the addition of 0.2 M
b) Lipid extraction by a mechanical process. phosphoric acid and hydrochloric acid. Similarly, the addition of 0.5%
acetic acid to water increases the recovery of acidic phospholipids. Fi-
5.2.1. Chemical method of extraction nally, they have concluded that Hajras method was the most efficient
5.2.1.1. Folch method. This is the most popular extraction technique, in procedure for extraction.
which solvents are used to homogenize the cells [77]. Several organic
solvents or mixture of different solvents have been recommended to 5.2.2. Mechanical method of extraction
selectively extract lipids from microalgae. The most common 5.2.2.1. Expeller press. The expeller press is considered as the oldest
combination used is chloroform and methanol in the ratio of 2:1 to a way for extraction of oil from algae. This is easy yet efficient
final volume twenty times (10 g sample in the 200 ml of solvent mechanical crushing process. The main principle involved in this
mixture) the volume of the algae cell for the extraction of lipids [78]. technique is, by applying high pressure the oil content in the dried or
After the addition of sample to the solvent mixture, the whole mixture wet biomass is out by breaking the cells. The application of the pressure
is exposed to gentle agitation (around 20–25 min) followed by should not be that much high, otherwise, the lipid quality, as well as the
centrifugation (2000 rpm) or filtration (with a funnel and folded filter quantity, gets decreased [84]. The expeller press works in the following
paper) to recover the liquid phase [78]. In some conditions, way; A screw type machine was used which presses the dried biomass.
dichloromethane also used instead of chloroform [79]. Fast and easy The dried biomass entered through one side and the product was exited
60
Table 4
Advantages and disadvantages of Folch and Bligh & Dyer methods.
Method Advantages Disadvantages
Folch Method • ItWellis aestablished

standard method. • Adverse effects of chloroform on the environment.
• Standard and simple method to determine lipids. • ItAdverse
is a laborious process/method.
Bligh and Dyer Method • It can determine totalmethod. • It is a laborious
effects of chloroform on the environment.
• Samples can be analyzedlipids. • process/method.
• directly without pre drying.
from the other side of the press. Continuous pressure and friction were is generated inside the vessel which results in the disruption of the cell.
applied for compressing the algae dried biomass. Upon applying the This will, in turn, enhance or open up the cell membranes of the
high pressure, the oil flows through the small openings which allow organisms with the electroporation effect. This technique involves
only the oil to flow through instead of allowing other components. quick generation of heat and high pressure inside the vessel resulting
During the application of a high-pressure temperature of 60–100ºC is in a good class of the extracts.
reached [84]. Hemwwimon et al. [91] and Šoštarič et al. [92] have revealed that
microwaves are the best pick nowadays due to short response period,
5.2.2.2. Bead beating. Bead beating method is another technique where less operational expenses, and capable extraction of algal oils. The re-
the algal cells are broken by the high-speed mechanism techniques vival time of biodiesel from the reaction blend is very less compared
[86]. The most common instruments used in this technique are the with the long process involved in the heating method which takes ap-
vibrating containers and agitated beads [87]. In the primary method proximately 6 h for completion of oil extraction [93].
(vibrating or shaking containers), the cells get damaged by shaking in
the complete container. Another technique followed in the bead beating 5.2.2.5. Electroporation technique. Hu et al. [94] explained that the
is the agitated bead method where the entire culture gets agitated [86]. lipid extraction efficiency from the cell walls of algae did not affect the
During this process, there is a lot of generation of heat which makes the composition and quality of extracted products such as fatty acids (FA).
instrument to release more heat. To reduce such heat, the instrument is From the algal biomass, maximum of 92% of the total lipid was
surrounded by the cooling jackets, hence it protects the sensitive extracted in a single electroporation treatment. On the other hand,
biomolecules during the process. Lee et al. [87] have shown that the only 62% was managed without the electroporation technique.
material of the bead inside the vessel also affects the lipid productivity
of the oil produced. Hence he had shown that optimal size for the cells
of the algae as well as the beads are in the size of 0.5 mm can increase 5.2.2.6. Osmotic technique. This is the best and effective technique
their hardness and density [87]. when compared to other extraction techniques. This technique can
disturb both the exterior as well as the interior of the algal cells in a
5.2.2.3. Ultra-sonication extraction. This is an alternative method for very fast and effective way [95]. Generally, in an osmotic press, there
the extraction which can be easily used. This process is furthermore are two different techniques which can be incorporated for extraction of
economical as well eco-friendly because it can be finished within a lipids.
shorter phase of time with high reproducibility [86]. The
Ultrasonication process involves the passing of sound waves which a) Hyperosmotic
propagate into the liquid which results in the alternating low and high b) Hypo-osmotic.
pressure generated or created. During this process, the bubbles
generated attain a specific size and collapse violently. This process is Basically hyperosmotic implies the concentration of the salt is
known as cavitation [88]. During the cavitation process, microbubbles higher outside the cells which cause damage to the cells. Conversely
are produced hence an ultrasound is applied which breaks the cell hypo-osmotic refers to the condition when the concentration of the salt
components of the algae. The key benefit of the ultra-sonication would content is low on the exterior and the fluid can flow easily into the cells
be that it utilizes low temperature in breaking the cell walls when to balance the osmotic pressure and the cells burst open if the stress is
compared with the heat generating equipment’s like autoclave, very high [95]. Among these two techniques, hypo-osmotic is fre-
microwave oven etc. [88]. Further, this technique does not involve quently used by many researchers to extract the inside components of
the chemicals or any other kind of stuff which in general are to be the cell. According to researchers, the osmotic pressure technique is the
removed later. The removal of these components is again a burden most simple, easy and most efficient way to extract the lipids from the
involving high cost as well as high labour. One of the recent microalgae species [96]. Several freshwater strains like Botryococcus
developments in the field of ultrasound extraction is the “ultrasound sp., Chlorella vulgaris, and Scenedesmus sp. have been tested and were
assisted extraction”. Generally, a frequency of 20–100 kHz is used for found successful, whereas research has to be conducted on the marine
common ultrasound processing. This frequency can be used to produce species and also this technique has yet to be tested in the pilot scale,
high acoustic pressure or acoustic cavitation. Different algal species where it is found to be unsuccessful [86].
differ in both their resistance to disruption and cell size [86].
5.2.2.7. Enzymatic assisted extraction. The cell wall structure of Algae is
5.2.2.4. Microwave extraction. The microwave extraction was first easily affected by the process of enzymolysis. It is a novel method of
demonstrated by Ganzler et al. [89]. They established a new method extraction of lipids from the walls of algal species. The enzymes like
for extracting the lipids from the food, algae, seeds etc. The application cellulose as well as trypsin are added to microalgal biomass [86]. The
process of this technique is quite simple when compared with other function of these enzymes is to break the tough cell wall of the algae
techniques. Amarni et al. [90] have explained the methodology of and extract the cellular components of them. The major disadvantage of
microwave extraction after a long time since Ganzler et al. [89] had this technique is that it should be conducted at a very low temperature
explained. In this process, a polar material is hosted in an electric field and the process is also very costly [97]. The extraction process can
which is oscillating very rapidly and produces microwaves which combine several enzymes which can degrade efficiently. The research
generate heat from the frictional forces which arise from the inter group had conducted an experiment where they had extracted oil from
and intramolecular movements. During this heating process, the vapour Scenedesmus species with the help of enzyme assisted hydrolysis [86].
61
Fig. 4. Different Products of algae through various extraction processes.
5.2.2.8. Lipid extraction by single step procedure. Axelsson and Gentili opportunity to increase the lipid inside the tiny algae for conversion of
[77] have proposed a method in which the algal material was made into oil and utilizing for biofuel production [99]. Researchers also con-
a paste and was suspended in 1:2 ratio of methanol and chloroform. centrate on the nutrient medium which is being used for growth culture
Later the tube was shaken vigorously up to the entire biomass was of algae [23]. The most researched species to date are Chlamydomonas
suspended in the solvent medium, 0.73% water and sodium chloride reinhardtii and Chlorella vulgaris. Now researchers also shifted con-
mixture were added to form a ratio of1:2:0.8 system of centration on the locally available species and those which do not have
methanol:chloroform: water respectively [77]. This technique was the lipid content. Genetic engineering and molecular biotechnology
effective in detecting that the one-step process which was found techniques are also being applied to such kind of species for increasing
suitable for extraction of total lipid and this process can be applied in the lipid productivity. By doing this the production of biofuel on a large
the selection of algae for qualitative and quantitative analyses of total scale would be much easier since there will be a mixed consortium of
FAs. Various extraction methods for different products have been algal species in the container or tank [23].
illustrated in Fig. 4.
The three main criteria which are considered in extraction steps in 6.2. Pollution control
improving the FA profile:
Nowadays pollution is a major threat from the harmful gasses
• Direct trans-methylation of lipids. emitted from the exhaust of vehicles as well as from the chimneys and
• Preliminary extraction steps were eliminated various power plants. These gasses are responsible for heating up the
• Fatty acid methyl esters (FAME) were generated by single step de- surface as well damaging the Ozone layer which makes the sun rays to
rivatization which helped to denature the protein fraction. directly pass through the atmosphere and causes severe damage to
human health [1,9]. Along with many plant species, algae also take part
6. Applications of microalgae in a significant role in cleaning the earth's atmosphere by absorbing the
CO2 for their growth. Microalgae function similar to plants and are
The usage of microalgae by humans was dated some 2000 years ago capable of photosynthesis. Using the energy from the sunlight they turn
by the Chinese people during the famine situation in their country. CO2 and water into sugar. Algae have a special enzyme in their cells
Nowadays there is a variety of commercial as well as industrial appli- with an ability to detoxify the nitric acid which helps the dangerous
cations of the algae (Fig. 5). With the genetic modification of the genes nitrate to get reduced and thus cleans the environment [101].
in potent organisms, we can achieve new products [98]. Other than
biofuel, microalgae have also been proven to produce important pro- 6.3. Wastewater treatment
ducts like pigments, polyunsaturated FAs, antioxidants, carbohydrates,
pharmaceuticals, natural colourants for cosmetics lipids, proteins and Nowadays along with pollution, wastewater released into many
too as an animal feed [12,23]. Algae is considered to be one of the most water bodies is also a major concern [102–106]. The water is not being
potential sources in each and every application of life sciences, some of purified properly and many human beings are affected by consuming
which include biofuel, bio fertilizer, bioelectricity, essential food sup- the partially purified water [107–112]. There are many techniques in-
plement, stabilizers, pollution control, wastewater treatment, biohy- volved in cleaning the wastewater with the help of algae. The most
drogen etc. [23,99,100]. prominent technique is the CO2 enhanced wastewater ponds, stabili-
zation ponds, anaerobic ponds etc. Research in this particular area has
6.1. Fuel sources proved that algae can survive the high level of toxic compounds in the
wastewater [113]. These toxic substances are utilized by algae for their
There is much amount of research done and still going on the bio- growth purpose and in turn, these magnificent species have shown that
fuel production from algae. Many researchers are utilizing this biomass and lipid content in the algal cells is increased, thereby giving
62
Fig. 6. Schematic design of single chamber microalgae catalyzed MFC.

Fig. 5. Schematic illustration of various potential applications of algae.
under most favourable parameters.
dual benefits. Further research should be done for the improvisation of
the techniques used in the treatment of wastewater [114,115]. Once the 6.7. Biomaterials and bioproducts
wastewater treatment infrastructure is developed in a full-fledged
manner, it provides an opportunity for biofilm technologies which is The macro-algae have their own kind of gel-formation and have the
also another key factor in the treatment of wastewater [116]. The capacity of dissolving in water because of which they are used in in-
biomass formed for the duration of the treatment of wastewater could dustries. Agar is a by-product of macro-algae, which has glutamic acid
be used as a potential source for the human as well as animal feed. which is a valuable chemical inside the cells [124].
6.4. Bioelectricity
6.8. Algae in cosmetics
Nowadays there is a great crisis of power supply affecting the daily
The extracts from microalgae are being used in cosmetic products
activity. More power is needed to empower gas turbines in generating
like anti-ageing and anti-irritants. They are also found in sunscreen
the required current [117]. Due to the continuous generation of current
lotions and hair care products. Few classic examples of commercially
from these sources, there is a heavy noise pollution and a lot of heat
accessible products are Arthospira which are rich in protein and are
generation during the process. Since these microalgae are very efficient
responsible for the repair of early skin ageing, tightening effect and
converters of solar power they can be used in producing the electricity
avoid stretch marks formation [124]. Chlorella vulgaris contains col-
through the biotic system as well as enhancing biomass [9]. Since it is
lagen which can stimulate collagen synthesis in the skin, which sup-
still an emerging process more exploration needs to be carried out in
ports the rebirth of the soft-tissue and reduce wrinkle formation. The
the field of bioelectricity to make a full-fledged algal-based microbial
component from D. Salina stimulates the cell propagation and enhances
fuel cell system to cut short the power crisis for the generations to come
the energy metabolism of the skin tissue [124].
[9,118–120] (Fig. 6).
6.5. Algal biohydrogen 6.9. Algae in human nutrition
Bio-H2 is a potential alternative source of energy. Researchers have Nowadays algae are being manufactured in the form of tablets,
considered microalgae as the easily and cheaply available source for the gums, capsules, syrups etc. Since they have a wide variety of chemical
production of bio-H2 [1]. Hans Graffon and his research team from composition they can be used in a variety of food products like pasta,
Germany have observed that the species Chlamydomonas reinhardtii snacks, gums, and drinks [23]. The four promising strains which are
which they were studying had frequently switched from the production used for this purpose are Arthrospira, Chlorella, D. salina and Aphani-
of O2 into H2 [121]. Later after some research, it was seen that the zomenon flos-aquae. The major reason for which Arthrospira is being
enzyme known as hydrogenase is responsible for the production of this used in a human diet is that it has an excellent nutritive value [124].
bio-H2. When the algal species are deprived of S they exchange on the
formation of H2 from O2 [122]. A less number of species are identified 6.10. Generation of stable isotopes by microalgae
for producing the bio-H2 [123].
Since algae are known to possess phototrophic growth capability,
6.6. Algal biogas they can be used for generation of precise molecules/substances which
are collectively labelled with 13C, 15N or 2H. The compounds from the
The major obstacle in generating the biogas from algal source is the above reaction can be utilized in generating many different products
feedstock related issues and also that most of them have seasonal var- [125]. Generally, drugs are developed depending on the interaction in
iation and needs to adjust to the ecological conditions [32]. The algal the middle of the receptor and ligand. The drug manufacturers study
biogas production has not been much explored and the various factors the interaction between the molecules among themselves and design
which could be considered are the pre-treatment of algal biomass [31]. the chemical structure of the drug. Hence the complex formation be-
It is also essential to investigate the effects of diverse pre-treatments tween these two is really advantageous in designing the drug.
63
Table 5
Different kinds of components synthesized by microalgae.
S.No Components Products Applications
Pigments/carotenoids Β-carotene Beta-carotene is a red-orange pigment, used as a precursor for vitamin-A

(retinol).
Astaxanthin Astaxanthin is a reddish pigment used for treating Alzheimer's disease.
Lutein and Zeaxanthin Lutein and Zeaxanthin are yellow to red pigments used to prevent eye
diseases.
Canthaxanthin It is a keto-carotenoid pigment which is widely used as lipid-soluble
antioxidant.
Chlorophyll Chlorophyll is a green coloured pigment which is used as a food additive
(colorant).
Phycocyanin It is a pigment-protein which is used as an antioxidant and anti-inflammatory
agent.
Polyunsaturated fatty acids Docosahexaenoic acid (DHA) (C22:6) DHA is a long-chain omega-3 fatty acid which is essential for visual and
(PUFA) neurological development
Eicosapentaenoic acid (EPA) (C20:5) EPA is a long-chain omega-3 fatty acid. It is a part of healthy diet
(recommended to reduce the risk of heart disease).
Arachidonic acid (AA) (C20:4) AA is omega-6 fatty acid which is a precursor molecule for the synthesis of
both the prostaglandins and leukotrienes in humans.
γ-Linolenic acid (GAL) (C18:3) GAL is omega-6 fatty acids used for the treatment of diabetics.
Vitamins Vitamin-A (retinol) Fat soluble and strong antioxidant. It is essential for immune function, vision,
reproduction, and cellular communication
Vitamin-B1 (thiamine) It is a part of an enzyme which help the body convert food into fuel.
Vitamin-B6 (pyridoxine) It is a part of an enzyme which help the body for protein metabolism and to
generate red blood cells.
Vitamin-B12 (cobalamin) It is a part of an enzyme which is vital for nerve function.
Vitamin-C (ascorbic acid) It is a part of enzyme required for protein metabolism and iron absorption.
Vitamin-E It functions as antioxidant.
Biotin It is a part of enzyme needed for energy generation.
Vitamin-B2 (riboflavin) It is essential for energy generation.
Vitamin-3 (nicotinic acid) It is used to prevent and treat of pellagra.
Vitamin-B5 (pantothenate) It is required for the synthesis of coenzyme-A (CoA).
Vitamin-9 (folic acid) It is a used to prevent and treat liver deseases, ulcerative colitis.
Antioxidants Catalases, polyphenols, superoxide dismutase, These are used as antioxidant supplementation.
tocopherols
Others Antimicrobial, antifungal, antiviral agents, toxins, These are having several medical, pharmaceutical, agriculture and industrial
amino acids, proteins, sterols. applications.
6.11. Breath test diagnostics distinctive features of marine algae in the presence of abundant
sulphated-polysaccharides in their cell walls. Another product from the
The family of microalgae may offer a means to build up a series of macro-algae which can be considered is the hydrocolloids. These can be
tests which are related to the gastrointestinal tract which are not only found in various red and brown seaweeds [124].
minimally invasive but also simple to quantify the sum of 13C in breath
CO2 [126]. These diagnostics tests are used as a replacement therapy
for the high-risk procedures (endoscopies and colonoscopies) [126]. 6.13.1. Ulvans
These 13C breath tests include the evaluation of the13C:12C ratio pre- These are the compounds extracted from the family Ulvales gen-
sent in breath CO2 when ingestion of a nutrient or supplementar- erally belonging to the genera Ulva and Enteromorpha. These are ex-
ymealhaving13C [127]. Some of the precise cases include the usage of tracted from water which contains a cation chelator from the cells of
labelled-galactose to observer liver function and the usage of labelled- green weeds [128]. The structure of these ulvans consists of various
xylose to conclude the range of microbial/bacteriological overgrowth of reiterating arrangements of rhamnose, glucuronic acid, iduronic acid,
the small intestine. Different kinds of components synthesized by mi- xylose, and sulphate [128].
croalgae are listed in Table 5.
Table 6
UV Screening compounds from microalgae.
6.12. High-value molecules
S. No UV-screening Species of algae
Pure molecules can be found in high concentration in many valu- compound
able products like fatty acids, pigments and sterols (Table 6). Since the
1. Sporopollenin Characium Terrestre, Coelastrum microporum, Enallax
plants and higher animals lack the system to generate or synthesize coelastroides, Scenedesmus Sp., Scotiella chlorelloidea,
polyunsaturated FAs [124]. Thus these FAs must be generated from the Scotiellopsis Rubescens, Spongiochloris Spongiosa,
food which we consume. Generally, fish and fish oils are taken to have Dunaliella salina and Chlorella Fusca
2. Scytonemin Chlorogloeopsis Sp., Calothrix Sp., Scytonema Sp.,
these FAs in the body, but due to the safety regulations and rules for the
Rivularia Sp., and Nostoc commune Lyngbya cf.
accumulation of toxins [23]. aestuarii Chroococcidiopsis Sp., Nostoc Punctiforme
3. Mycosporine-Like Ankistrodesmus spiralis, Chlorella minutissima,
6.13. Small molecules Amino Acids Chlorella sorokiniana, Dunaliella tertiolecta, Scotiella
Chlorelloidea, Isochrysis Sp., Pavlova gyrans, Corethron
criophilum, Thalassiosira tumida, Porosirapseudo
Since many years’ macro-algae has been employed for the produc-
Denticulata, Stellarimamicrotrias, Thalassiosira
tion of alginates, carrageenans or agars, which are either located in cell weissflogii, Alexandrium catenella
walls or within the cells and serve as storing materials [23]. One of the
64
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Renewable and Sustainable Energy Reviews 94 (2018) 49–68

Contents lists available at ScienceDirect

Renewable and Sustainable Energy Reviews

Production of biofuels from microalgae - A review on cultivation, T

Fig. 1. Classiﬁcation of algae explained in a simple way.

• Algae contain a high amount of iron (Fe) which is advantageous if

Nitrogen is one of the most plentiful sources available in nature next

Biomass yield Lipid production Total lipid

Strain Habitat Nutrients Biomass Lipid Ref.

Biomass yield Lipid production Total lipid

Strain Habitat Nutrients Biomass Lipid Ref.

Biomass yield Lipid production Total lipid

Strain Habitat Nutrients Biomass Lipid Ref.

Biomass yield Lipid production Total lipid

Note: n.a – not available.

1. Alum Polyvalent 5.3–5.6 Wastewater [169]

• The simplicity of manufacturing and functioning

5.1.2.6. Electrolytic process. In this process, the movement of

• Medium handling of huge quantity of samples is the key advantage of this

Folch Method • ItWellis aestablished

Fig. 4. Diﬀerent Products of algae through various extraction processes.

Fig. 6. Schematic design of single chamber microalgae catalyzed MFC.

6.5. Algal biohydrogen 6.9. Algae in human nutrition

Pigments/carotenoids Β-carotene Beta-carotene is a red-orange pigment, used as a precursor for vitamin-A

7. Conclusion extraction and evaluation of single-step biodiesel production. Eng J

• Production of algal biodiesel necessitates huge-scale cultivation and

Oceanogr 1978;23:283–95. of electro-coagulation-ﬂocculation for harvesting marine and freshwater micro-

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