Biocatalysis and Agricultural Biotechnology 25 (2020) 101580

Contents lists available at ScienceDirect

Biocatalysis and Agricultural Biotechnology

Microalgae – A green multi-product biorefinery for future industrial

prospects
Munna Bhattacharya, Saswata Goswami *
Center of Innovative and Applied Bioprocessing, Knowledge City, Sector-81, Mohali, Punjab, 140306, India

ARTICLE INFO ABSTRACT

Keywords: Microalgae offer fascinating highlights to be enable them as elective raw material for different applications
Microalgae through bio-refining process. They have special capacities to use them for biotechnology-oriented applications.
Biorefinery
In this way, this review examines to grow the extent of incorporating microalgae along with different other
Value-added products
biorefinery applications to upgrade their maintainability. The utilization of microalgae as animal and fish
Techno-economic analysis
Opportunities and constraints
feed, manures, pharmaceutical, cosmeceutical, environmental and other biotechnological applications are alto-
gether assessed. It additionally features the boundaries, openings, improvements, what's more, probabilities of
expanding the extent of microalgae through techno-economic analysis. This review infers that subsidizing sup-
ported research and a move of microalgal research objective from biofuels generation to bio-refinery co-items
can ensure them as valuable resource. In addition, innovation joining is inescapable to bypass the expense of
biomass handling of microalgae. It is anticipated that this review would be useful to decide the future job of
microalgae in bio-refinery applications.

1. Introduction and are particularly interesting for pharmaceutical industries to de-

Microalgae contain high measure of proteins, lipids and complex
sugars that can be utilized as feedstock for generation of various bio-
products. They are developed in open lakes or culture ponds in closed
systems using just water and environmental CO2, which might be ac-
cessible at least expense. In any case, a culture medium enriched with
supplements and salts that is essential for microalgae culture develop-
ment (Baicha et al., 2016), yet the processes are less energy consum-
ing, hence the main focus is maximum utilization of microalgae with
least energy consumption (Vanthoor-Koopmans et al., 2013). Other
than that, biomass of microalgae likewise has high rates of photosyn-
thetic activities, which when combined with bioproduct generation
frameworks, has the capacity to be an economical pathway to get sus-
tainable energy generation hotspots for the future as reported by
Khoo et al. (2013). Separated lipids from microalgae can be utilized
as a vital resource for generating biodiesel whereas the carbohydrate
portions can be transformed to bioethanol in brewing industries, so
that the plant based customary carbohydrate sources like treated lig-
nocellulosic biomass or simple sugars can be replaced. Besides, long
chain unsaturated fats, microalgae derived proteins and pigments
have remarkable abilities as health promoting nutritional supplements
velop products for wellbeing sustenance and to treat several illnesses
(Yen et al., 2013). The essential job of microalgae that proves its sig-
nificance to be taken into account as an alternate option in contrast to
numerous regular segments and sources from plant origin are the dual
advantage of production of biofuels and several value-added bio-
based products. What's more, microalgae can biosequester CO2 from
environmentally hazardous gases produced from power generation in-
dustries and this will add to significant decrease of ozone harming
substances discharges thus preventing global warming (Cheah et al.,
2015).
Microalgal biomass has as of late turned out to be progressively
huge as an elective hotspot for sustainable energy generation. The
scan for sustainable energizes has picked up consideration because of
the increasing demand of energy and expanding total world populace
over decades. Some methodologies such as hydrothermal liquefaction
have been recommended for extraction of oils utilizing microalgal
biomass which could enlarge the market possibilities of microalgae
based bioproducts and also attracts further potential outcomes from
dual benefit of coupled production of algal biofuels along with value
added products. Therefore, increased consideration inclined to the us-
age of microalgae in different areas comprising food, energy, chemical

* Corresponding author.
E-mail address: saswatagoswami2015@gmail.com (S. Goswami).

https://doi.org/10.1016/j.bcab.2020.101580
Received 3 December 2019; Received in revised form 18 March 2020; Accepted 19 March 2020
Available online 23 March 2020
1878-8181/© 2020 Elsevier Ltd. All rights reserved.
M. Bhattacharya and S. Goswami
feed and pharmaceuticals are currently in progress. Phototrophic mi-
croalgae are at present utilized at industrial scale for the generation of
a few value-added or specialty products. Instances of these products
are lipids or oils as fuel; pigments, for example, astaxanthin and beta-
carotene; and dissolvable proteins, for example, phycobilliproteins
(Guo et al., 2013), which are presently developed commercially using
a few number of microalgal species including Haematococcus pluvialis,
Chlorella vulgaris, Dunaliella salina, and the cyanobacterium Spirulina
platensis (Yen et al., 2013). Nonetheless, right now just a single ex-
plicit compound (e.g., astaxanthin, b-carotene) is separated, while
other conceivably significant parts (e.g., proteins, carbohydrates) are
disposed off, otherwise underestimated.
The idea of biorefinery is like conventional oil refinery, with the
end goal that biomass is changed over into profitable fuels, chemicals
or compounds as reported by Pérez et al. (2017). The fundamental
contrast among oil refinery and biorefinery is as far as the crude feed-
stocks such as unrefined petroleum or biomass are concerned along
with the utilized technology (Moncada et al., 2015). By generating
both bulk and high value compounds from the developed algae, the
complete monetary gain can be increased from algae (Moncada et al.,
2015). Closely resembling an oil/petrochemical refinery, the microal-
gal biomass ought to be actualized for complete fractionation and val-
orization and the idea is conceptualized as microalgae associated
multi-product biorefinery (Jacob-Lopes et al., 2015). Such a facility
commonly comprises several steps such as culturing, harvesting and
disruption of cells, and product isolation steps. Additionally, separa-
tion or extraction steps must be completed to fractionate or purify the
product portions such as, segregation of the carbohydrates from the
aqueous stage to separate out water soluble proteins.
This review expects to outline the latest data about the standards
and learning on biorefinery of microalgae for the high value product
generation. The essential standards and basics about biorefinery of
microalgae are checked on and both the upsides and downsides of
these procedures are assessed. The product profile investigated in cur-
rent survey incorporate proteins, carbohydrates, lipids, pigments, an-
tioxidants, polyunsaturated unsaturated fats, vitamins, nanocellulose
and bioplastics. This survey additionally considered the probability
Fig. 1. Flow diagram of a microalgae biorefinery in summarized form.
2
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
for biorefinery of microalgae regarding monetary assessment and
maintainability as well as opportunities and constraints.
2. Microalgae as a multi-product biorefinery
Microalgae are delegated potential candidates in biorefinery forms
since they are able of delivering a handful of valuable products
(Sharma et al., 2017). They are considered as sustainable resource of
biomass, which is useful as far as fast development, diminished spe-
cific challenge regarding with food supply chain. The fractions of oil,
minerals, starch and proteins could be utilized in generation of high
value products such as biochemicals, biofuels, feed, biogas and other
important bioproducts. In process digestates, for example, glycerine
can likewise be changed into value added products also. Proteins and
carbohydrates part that are relatively low value segments may have
the ability to be changed into energy through consolidated heat and
power generation in the biorefinery. The overall biorefinery process
can be summarized as shown in Fig. 1.
The transformation innovations used in for microalgae biomass
can be separated into four classifications, to be specific conversion
through thermochemical reactions, conversion through biochemical
reactions, conversion through transesterification and photosynthetic
conversion in microbial fuel cell (Baicha et al., 2016; Mathimani and
Pugazhendi, 2019). Thermochemical conversion employs the standard
of heat deterioration of natural compounds present in biomass to ex-
tricate fuel items. Instances of thermochemical transformation forms
incorporate gasification (production of syngas), thermal liquefaction
(production of bio-oil), pyrolysis (production of bio-oil, syngas, char-
coal etc.) and direct burning or combustion (generation of heat and
electricity). The biochemical transformation represents the natural
bioprocessing of biomass for transforming energy into biofuels. In-
stances of biochemical transformation forms incorporate anaerobic di-
gestion (production of biogas), alcoholic fermentation (production of
bioethanol) and photobiological hydrogen generation (production of
biohydrogen). Biodiesel can be obtained through transesterification
reaction in which the triglycerides of algal lipid constituents react
with alcohol within the sight of a catalyst. This procedure can deliver
fatty acid methyl esters (FAME) and glycerol by utilizing methanol
M. Bhattacharya and S. Goswami Biocatalysis and Agricultural Biotechnology 25 (2020) 101580

and ethanol and the responses of triglycerides to FAME and glycerol
are typically catalysed by the reaction of an acid or base. For total
lipid conversion to biodiesel, generally a homogeneous or heteroge-
neous synergist process or even super critical carbon-dioxide extrac-
tion process have been utilized as per literature. Another important
aspect are the microbial fuel cells which are basically bio-
electrochemical gadgets that are capable for producing power or bio-
electricity from the biodeterioration of natural compounds under
anoxic conditions. The microalgae incorporation in microbial fuel cell
have found immense potential in oxygen production too, due to the
expulsion of CO2 by photosynthesis process through green algal
growth (Pradhan et al., 2017; Soares et al., 2013).The initial parame-
ters influencing the decision of process of transformation are the
amount and kind of feedstock raw biomass, monetary contemplations,
detail of tasks and the desired product prototype.
Biorefineries from microalgae have built up the advancement of
change of biomass into fuels, beauty care products, synthetic chemi-
cals, food, feed what's more, high value products (Wang et al., 2015a,
b; Vanthoor-Koopmans et al., 2013). The microalgae-based carbohy-
drates comprise for the most part of cellulose and starch with very
less lignin or without lignin, which acts as promptly accessible carbon
source for the brewing industry just as biobutanol and bioethanol
preparations. A few microalgae, for example, Nannochloropsis sp,
Tetraselmis sp, Isochrysis sp, Thalassiosira sp. what's more, Chaetoceros
sp, can likewise deliver long chain unsaturated fatty acids, for exam-
ple, DHA and EPA, which are profitable as nutritional supplements.
What's more, microalgae derived proteins and pigments have signifi-
cant imminent for different medicinal applications.
Microalgae show incredible potential for energy generation from
sustainable resources by means of biochemical forms without compro-
mising the food security and agricultural issues. However, this makes
the requirement for revenue generation for each region of land con-
trasted with different yields, less water utilization, maximum oil con-
tent and the capacity to be developed in arid lands also. Significant
intrigue has been raised for microalgae based biofuel generation just
as synthetic chemicals, food and pharma industries (Jacob-Lopes et
al., 2015). Additionally, ongoing investigations have shown that the
present advancements in economic situations what's more, innovation
in biochemical technology have made the generation of microalgae
biofuels financially practical.
3. High value products from microalgae and their industrial
applications
Microalgae are commonly unicellular autotrophic organisms nor-
mally can be traced in fresh water and sea water both. Complex com-
pounds are being produced for example, lipids, carbohydrates, and
proteins, utilizing any carbon rich substances found in environment.
They utilize normally the carbon-di-oxide from air for their growth
and energy production. There are some microalgae which can utilize
organic carbon source to produce biomass also (Hochman and
Zilberman, 2014). According to the taxonomic data, there are more
than 300,000 species of microalgae out of which around 30,000 are
archived (Richmond and Hu, 2013). They prefer to inhabit in complex
natural habitat and can adjust quickly in outrageous conditions such
as change in salinity, temperature, nutrients, light intensity etc. Sub-
sequently, they can produce an extraordinary assortment of intriguing
secondary metabolites as bioactive molecules having complex confor-
mations with biological efficacy that are for the most part not found
in different organisms (Herrero et al., 2013; Anbuchezhian et al.,
2015). The commercial development of microalgae has started five
decades prior and Japan was the first country to present business ap-
plications of microalgal species Chlorella in 1960s. The worldwide
market estimation of small scale algal growth is assessed to be around
US$6.5 billion, out of which about US$2.5 billion is generated by the
health food segment, US$1.5 billion by the generation of DHA and
US$700 million by aquaculture. The yearly productioin of microalge
is around 7.5 million tons (Sathasivam et al., 2019). Some valuable
bio-products that have been produced by microalgae are carotenoids
(particularly β-carotene), astaxanthin, docosahexaenoic acid (DHA),
eicosapentaenoic acid (EPA), bioactive and natural pigments, natural
dyes, polysaccharides, cancer prevention agents and algal concen-
trates. In the course of the most recent 20 years, the applications of
microalgae are pointed towards the use of four noteworthy microal-
gae: a) Spirulina (Arthrospira), b) Chlorella vulgaris, c) Dunaliella salina,
and d) Haematococcus pluvialis. Table 1 features on a few promising
microalgal species, their high-esteem items and application.
3.1. Lipids
Microalgae amass various lipids, that typically contributes up to 3–
20% of their dried biomass under ideal development conditions. As
indicated by the carbon chain length, lipids are grouped into two
sorts: (i) fatty acids having chain length of 14–19 carbon atoms, that
would changed over into biodiesel; and (ii) PUFAs with in excess

Table 1
Some major microalgae species, their compositional analysis, products and applications (Adapted from Koyande et al., 2019).

Species/group Composition (% dry matter) Product Areas of application

Protein Lipid Carbohydrate

Arthrospira (Spirulina) platensis 46–63 4–9 8–14 Phycocyanin, biomass Health food, cosmetics
Arthrospira (Spirulina maima) 60–71 6–7 13–16 Protein, Vitamin B12 Antioxidant capsule, immune system
Aphanizomenon flos- aquae 62 3 23 Protein, essential fatty acids, β- carotene Health food, food supplement,
Chlorella spp. 57 2 26 Biomass, Carbohydrate extract Animal nutrition, health drinks, food supplement
Chlorella vulgaris 51–58 14–22 12–17 Biomass, Carbohydrate extract Health food, food supplement, feeds
Dunaliella salina 57 6 32 Carotenoids, β- carotene Health food, food supplement, feeds
Haematococcus pluvialis 48 15 27 Carotenoids, astaxanthin Health food, pharmaceuticals, feeds
Odontella aurita 39–61 22–38 14–18 Fatty acids, EPA Pharmaceuticals, cosmetics, anti- inflammatory
Porphyridium cruentum 28–39 9–14 40–57 Polysaccharides Pharmaceuticals, cosmetics
Isochrysis galbana 50–56 12–14 10–17 Fatty acids Animal nutrition
Phaedactylum tricomutum 38–42 18–20 20–25 Lipids, fatty acids Nutrition, fuel production
Lyngbya majuscule 25–33 11–13 28–37 Immune modulators Pharmaceuticals, nutrition
Scennedesmas spp. 47 1–9 21–52 Protein Aquaculture, human nutrition
Schizochytrium sp. 40–46 22–24 10–14 DHA and EPA Food, beverage and food supplement
Crypthecodinium cohnii 28–45 22–38 25–33 DHA Brain development, Infant health and nutrition
Nannochloropsis oculata 63 11 15 Biomass Food for larval and juvenile marine Fish
Nannochloropsis sp 42–50 12–16 28–30 EPA Food supplement and pharmaceuticals

3
M. Bhattacharya and S. Goswami
chain length of 19 carbon atoms, which are most widely recognized as
microalgal PUFAs for eg. eicosapentaenoic acid (EPA) and docosa-
hexaenoic acid (DHA). These ω– 3 lipids are of specific enthusiasm for
application as wellbeing sustenance supplements because they are
viewed as fundamental in human nutrition (Lai (2015). EPA/DHA
capsules which contains lipids extracted from fish oil and can be pur-
chased in drugstores and pharmacies for a couple of US dollar. How-
ever, while fish oil prices are continually expanding and are antici-
pated to increment further (http://www.oecd.org/site/oecd-
faoagriculturaloutlook/highlights-2013-EN.pdf), worldwide interest is
likewise developing. In this manner, PUFAs derived from microalgae
bestows a practical and vegetarian elective to substitute fish oil and
may fill a market-specialty in the medium-value product segments.
Besides, it can't be rejected that algal oils by and large can enhance
the vegetable oil market to a high degree. Notwithstanding PUFAs,
other lipids from microalgae can be utilized for industrial purpose as
chemical compounds for example, waxes for the cosmetics industry
(Michalak et al., 2015), ointments or on the other hand fabricating lu-
bricating components for polymers.
3.2. Proteins
The development of the worldwide populace alongside expanding
interest for poultry and dairy items will increase the pressure on ani-
mal feed for protein supply. The Food and Agriculture Association of
the United Nations predicts an ascent of worldwide meat utilization to
300 million tons in 2020 (from 233 million tons in 2000). So to reach
out the needs in future, researchers take up the challenge to sort out
alternate options to the available sources of protein. In such manner,
microalgal proteins provide fantastic wholesome quality by conveying
every single amino acid, comprising all basic amino acids (Lum et al.,
2013). In the long haul, microalgae could be utilized for human nutri-
tion just as animal feed.
3.3. Carbohydrates
Carbohydrates are the primary component that can contribute
over half of complete dry weight of biomass and for the most part fill
in as storage and structure molecules (Arad and Levy-Ontman 2013).
The microalgal carbohydrates are for the most part starch and cellu-
lose, which can provide the source of feedstock for bioethanol produc-
tion. Starch derived from microalgae could likewise be used as renew-
able raw material for fermentation instead molasses derived from sug-
arcane, for that reason and subsequently contribute to sparing agricul-
tural land and protection of environment. In comparison to plants, mi-
croalgae does not have complex structures, for example, their cell
wall does not comprise of hemicellulose or cellulose fibrils bound
with lignin, however microalgae contains a wide range of and increas-
ingly complex intracellular structure and exhibit fascinating extracel-
lular polysaccharides, particularly sulphated polysaccharides, which
are in charge of quite certain biological efficacies. Sulphated polysac-
charides are highly bioactive, which enables them for the generation
of bioactive materials, beauty care products, nutritional and pharma-
ceutical applications (Arad and Levy-Ontman 2013). Besides, water-
soluble lubricants and thickening agents like agar can also be synthe-
sized utilizing microalgal polysaccharides (Sathasivam et al., 2019).
3.4. Pigments
Three fundamental types of common microalgal pigments ought to
be accentuated are carotenoids, chlorophylls and phycobiliproteins.
Most of such pigments have been recognized as vitamin precursors in
food and nutrition but also extends their application to animal feed,
food additives, cosmetics, pharmaceuticals, food coloring agents and
biomaterials (Sharma et al., 2017; Nobre et al., 2013). Among the mi-
4
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
croalgal pigments, carotenoid pigments are lipid-soluble pigments that
offer coloring to different plant parts and also known as accessory pig-
ments. Generally, extraction strategies for microalgae derived
carotenoids utilizes Soxhlet extractions with organic solvents (Chen et
al., 2016), but super critical fluid extraction is now the most popular
where the determination of solvent is huge to decide the level of affin-
ity and speed up the carotenoid extraction. Astaxanthins are one of
the pigments, which have potential antioxidant activities such as anti-
aging, anti-inflammatory and immune enhancing effects that can be
utilized for food and nutraceutical industries (Cheng et al., 2016). The
other pigment, chlorophylls are also soluble to lipids having low ex-
tremity, that are generally present in vegetables and natural products
as essential pigments for photosynthesis. Like carotenoids, the con-
ventional technique for removing chlorophyll requires a few extrac-
tion steps with organic solvents. A number of variables in solvent ex-
traction may influence the pigment yield, to be specific the sort of sol-
vent utilized, procedure for cell wall disruption, extraction time and
utilization of various empirical correlations (Assemany et al., 2015).
In microalgae, phycobiliproteins are the abundant accessory pigments
that helps in photosynthesis process. The significant uses of phyco-
biliproteins incorporate characteristic colors in food commodities and
pharmaceuticals items. It displays cancer prevention agent, antiviral,
antioxidants, anti-allergic, hostile to hypersensitivity, anti-
inflammatory and neuro-defensive properties which shows its poten-
tial in healthcare related applications (Chen et al., 2013; Chang et al.,
2015). Centrifugation, drying, homogenization, repeated freeze thaw
process are continuously used to extract phycobiliproteins from
miroalgae (Tiwari et al., 2019).
3.5. Vitamins
In contrast with the commodity food items, elevated amounts of
fundamental vitamins and trace elements are found in microalgae,
which also shows substantial bioavailability. The bioavailability stud-
ies on these compounds are frequently needed contrasted with the
quality of the feed, so that the impact of these segments when brought
into the body might be assessed. Microalgae needs nitrogen (N) acces-
sibility for the production of essential vitamins. They found that the
cyanobacteria delivered less quantity of vitamin B12 per cell in low-N
conditions contrasted with N-loaded conditions. The percentage of vit-
amin yield was likewise influenced by the nitrogen source and also its
concentration in the culture medium. Riboflavin is another vitamin,
that are also can be derived from microalgae, that serves as a nutrient
fundamental for maricultured life forms.
3.6. Polyunsaturated fatty acids
Polyunsaturated fatty acids (PUFA) are broadly perceived as fun-
damental health promoting supplements that aid the counteractive ac-
tion of different heart ailments. These compounds can be recuperated
from sea fish, but now has been discouraged by the quick consump-
tion of marine resources, where expanding demands for PUFAs have
spurred the revelation of alternative resources of EPA and DHA. In
this regard, microalgae are naturally equipped for PUFA production
which are critical for human nutrition (Wang et al., 2015a,b). Ad-
vances in microalgae innovation can be implemented for the genera-
tion of PUFAs. This is because microalgae is a sustainable source and
can be cultured throughout the entire year with high oil substance
and vast biomass generation. A few techniques incorporated Bligh and
Dryer extraction, solvent extraction and sonication, direct saponifica-
tion furthermore, supercritical fluid extraction (SFE) for fatty acid
analysis in marine microalga with comprehensive results (Li et al.,
2014).
M. Bhattacharya and S. Goswami
4. Valuable by-products from microalgae biorefinery
In the contemporary world, the weather change as a result of mas-
sive consumption of petroleum derived products has praised the con-
sumers to employ algae as source for generating bioenergy. The aim
of the work is to enhance the energy production and increase the ac-
cess of energy resources to every mankind. The term biofuel signify to
any solid, liquid, or gaseous material that can be used for energy pro-
duction, derived from living or dead biomass. However, the total
amount of energy generated relies on nature of biomass, its quantity
used as substrate and procedure applied for the production of the en-
ergy (Vigani et al., 2015); Jacob-Lopes et al., 2015). The agricultural
crops are regarded as the first generation biofuels being the source of
their utilization for food or feed. Nevertheless, the type of food mater-
ial stuff determines the quantity of fuel produced over time. In con-
trast with biofuels derived from agriculture residue feedstock, the al-
gal biomass production does not involve farming. The algal biomass
production efficiency can be improved via selection of suitable algae,
use of cheaper source like wastewater treated products and, altering
the chemical structure and oil content of algal biomass.
4.1. Bio-oil
Bio-oils also termed as biocrude oil, pyrolysis oil, obtained by ther-
mal treatment of dried biomass in the absence of oxygen at a temper-
ature about 500 °C and then subsequently subjecting it for cooling.
They are similar to petroleum derived oils in terms of generating en-
ergy and thus can be employed as its substitutes (Zhang et al., 2015).
The production of bio-oil can be classified into two stages: pyrolysis
and thermo-chemical liquefaction (Tommaso et al., 2015). Pyrolysis is
generally performed at high temperature (350–530 °C) to convert
dried biomass into liquid, solid and gas. The liquid part consist of bio-
oil or tar and biomass. Further it contains different natural com-
pounds including lipids, proteins and carbohydrates. However, the
amount of biomass production is proportional to lipid content in the
present in the algae. Many microalgae have been explored for the pro-
duction of bio-oil by the process of pyrolysis or thermal liquefaction
method (Pan et al., 2018). Some of them includes Spirulina (up to
41%), Scenedesmus (24%–45%), Dunaliella (about 37%), Desmodesmus
(up to 49%) (Godwin et al., 2017).
4.2. Biodiesel
In past few years, oil seed crops like soybean oil, palm oil and
rapeseed oil have been reported to be used as source for the biodiesel
production (Godwin et al., 2017). The production cost of biodiesel
mainly depends on the sort of crude material utilized representing 50–
85% of total fuel cost. However, for the cost-effective biodiesel pro-
duction, utilization of feedstock is crucial parameter that could be em-
ployed (Selvaratnam et al., 2015). Usually, the biodiesel id produced
by transesterification reaction that involves the conversion of tria-
cyleglycerols or free unsaturated fats into non-toxic products consist-
ing. The refinement of algal oil is of prime importance due to conver-
sion of high molecular weight oil into low molecular weight com-
pounds such as fatty acid alkyl esters. The two algal species Chlorella
vulgaris and Chlorella protothecoides have been reported by many re-
searcher for the presence of high oil content in their cell. Gülyurt et
al. (2016) assessed the capability of Chlorella protothecoides for
biodiesel production by microwave-assisted transesterification
process. The reaction was carried by using oil produced from Chlorella
protothecoides and methanol. Potassium hydroxide was used as a cata-
lyst. The proportion of methanol and oil, and catalyst and oil, the re-
action time were used as the procedure parameters that influenced the
yield of methyl ester. From the carried experiments, 9:1 methanol:oil
molar proportion, 1.5% KOH catalyst:oil proportion and 10 min were
5
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
ideal conditions for biodiesel production. However, the biodiesel pro-
duced from microalgae is generally carried from unsaturated fats
(Sharma and Sharma., 2017). The algal biomass derived from waste-
water contains a mixture of different alga and subsequently unique
fatty acid profiles can be acquired (de Souza et al., 2019) reported the
production of biodiesel from Chlorella sp., Euglena sp., Spirogyra sp.,
Scenedesmus sp., Desmodesmus sp., Pseudokirchneriella sp., Phormidium
sp., Nitzschia sp..
4.3. Biohydrogen
The need for renewable energy sources raised attention towards
bio-hydrogen production. Despite of many efforts, the production of
bio-hydrogen on large scale did not get success because of high pro-
duction cost, involvement of sophisticated machineries, and low bio-
mass fixation. Many researchers have reported the production of
biodiesel by exposing certain green algal species to different stress
conditions such as deprivation of light that could trigger apparent
yield of hydrogen gas. To study the process and parameters utilized
for hydrogen gas production from algal biomass, numerous reports
were studied (Saifuddin and Priatharsini, 2016; Show et al., 2018).
There are three main pathways for generation of hydrogen- (1) Direct
photolysis, (2) Indirect photolysis, and (3) ATP driven pathway. Algae
use photosynthetic capability or direct photolysis to convert water
into oxygen and hydrogen, resulting in the. However, the simultane-
ous production of hydrogen and oxygen may results in more security
dangers expanding the total cost of production process involves. Fur-
thermore, the reaction is catalysed by hydrogenase enzymes which is
prone to get damaged in the presence of oxygen as it is oxygen sensi-
tive. Because of these disadvantages, indirect photolysis is more rec-
ommended over direct photolysis method for hydrogen production.
Many reports are present to reveal the generation of hydrogen under
anaerobic and sulphur restricted condition, where the carbohydrates
present in the algal cell wall changes into hydrogen. In the majority of
the studies, cyanobacteria are the principle producers of biohydrogen
through biologically driven ATP pathway, utilizing hydrogenase and
nitrogenase enzymes as catalysts (Saifuddin and Priatharsini. 2016).
4.4. Biomethane production
Now-a-days, the production of biogas from biomass is getting im-
portance due to need of consumers and with awareness among the
users for the use of eco-friendly sources of energy. The biogas produc-
tion can be carried out in an anaerobic digester containing synergistic
microbial population that convert algal biomass compounds (lipid,
protein, sugar) to methane and carbon dioxide. Then methane can be
efficiently utilized for fuel as well as for chemical feedstock (Leu and
Boussiba, 2014). In a report of Uggetti et al., (2016), where they had
examined the capability of various feedstocks, for instance algae,
wood, grass and solid waste, for biomethane production. They ob-
served that green algal biomass served as a potential contender for
generation of biomethane. The biomass productivity of algae is gener-
ally higher than the terrestrial plants however, the development is
limited due to many factors. Recently, Wang et al. (2016) studied a
method to enhance biomethane productivity from microalgae -
Chlorella sp, by pre-treating with heat. The biomethane production in-
creased from 37% to 48% as the temperature is increased form 70 °C–
90 °C for 0.5 h as compared with control algal biomass.
4.5. Bioethanol production
The utilization of algal biomass for the production of bioethanol is
acquiring attention because of many factors including high biomass
efficiency, better variety of algae, presence of extensive composition
of starch and carbohydrate, thin cell wall structures, and have high
M. Bhattacharya and S. Goswami
photosynthetic rates of the organisms. Normally, production of
bioethanol from algae involves two methods-fermentation and gasifi-
cation (Uggetti et al., 2016). In different countries, the production is
done by large scale fermentation of sugary and starchy agricultural
crops. The agricultural biomass is usually crushed or grinded, and
then the starch present in the crops get converted into sugars with the
help of different enzymes, yeast (mostly Saccharomyces cerevisiae),
acid and alkalis. Audu et al. (2018) had investigated the capability of
marine green alga Chlorococcum littorale to produce ethanol under
dark fermentation. They found that about 27% of starch present in al-
gal cells was utilized within 24 h at 250 °C for the bioethanol produc-
tion. Sharma at al. (2017) evaluated the feedstock capability of algal
biomass to deliver bioethanol and to encourage its use as sustainable
biofuel for giving legitimate choice. Many microalgae are used for
bioethanol production such as Dunaliella, Chlorella, Chlamydomonas,
Arthrospira, Sargassum, Spirulina, Gracilaria, Prymnesium parvum, Eu-
glena gracilis and Scenedesmus. Among the different types of algae such
as brown, green, red algae, brown algae is considered to be potential
stuff for the bioethanol production due to the presence of high carbo-
hydrate content in the algal cell and ease of culture development
(Jung et al., 2013). The fermentation of laminarin (produced from
Laminaria hyperborean) by the yeast-Pichia angophorae produced ade-
quate mount of bioethanol (Meinita et al., 2013). The red algae have
also been reported for the bioethanol production using acid hydroly-
sis, but the yield was very low (approximately 45%). The seaweeds
are also reported to produce bioethanol in the range of 0.08 and
0.12 kg per kg of dry seaweed, depending upon the depending upon
the genera of algae and different methods for pre-treatment and hy-
drolysis (Shimakawa et al., 2016). Another group of workers pro-
duced a potential yield of ethanol roughly about 90 L per tonne of
dried macroalgae, however the yield was low as compared with land
based harvests for biofuel generationtechnology is microfiltration,
which is a low energy . Puschner (2018) from their investigation have
gone to a point that creation of ethanol from dark colored ocean
growth needs higher ethanol yields for modern procedure and utiliza-
tion of all elements of kelp for non fuel based items moreover. In
Brazil, bioethanol is commercially utilized as pure ethanol or blend of
ethanol with oil used in about 86% of sold vehicles. However, at the
same time, there are many demerits in the production process as they
have low vapour pressure, low energy density and low fire retardancy
(Puschner, 2018).
4.6. Biobutanol production
In many countries of Asia, Europe and South America, algal cul-
ture development is being carried out for bioethanol and biogas gen-
eration, though in USA, they are used for biobutanol production. For
past 100 years, butanol has been utilized as a fuel in transportation
and has been suggested as a potential source of biofuel, not exclu-
sively to extemporize, however to substitute of ethanol as petroleum
additive substance because of its low vapour pressure and high energy
density (Cho et al., 2017). The microbes used for butanol production
uses starch, sugars, cellulose present in algal biomass as substarte;
consequently butanol production could be as monetary profitable as
ethanol (Gao et al., 2016). By anaerobic fermentation, numerous
Clostridium sp. are capable for the production of acetic acid, butanol
and ethanol (ABE) by using the two hexoses and pentoses sugars by a
procedure called as ABE fermentation (Wal Hetty et al., 2013). How-
ever, the butanol inhibits fermentation process prompting less yield
and productivity. Clostridium used Ulva lactuca as substrate material
for the butanol production by fermentation. The substrate was ini-
tially utilized pre-treated with hot water, then subjected for enzymatic
hydrolysis by commercial cellulases to produce acetic acid, butanol
and ethanol (ABE). The final yield of 0.35 g ABE per gm of carbohy-
drate content was obtained (Wal Hetty et al., 2013). Hence these re-
6
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
sults showed the capability of Ulva lactuca as a feedstock for ABE fer-
mentation. However, in between the process, there was generation of
1, 2 propanediol (propylene glycol) as a by-product limiting the total
volume of the butanol. Macrolagae Ulva extricated from Jamaica Bay,
New York City, was utilized for productioin of butanol on a pilot scale
from saccharification of sugars (Cho et al., 2017).
4.7. Algal bioplastics
Algal biomass can be used for the production of different forms of
plastics such as-hybrid plastics, cellulose-based plastics, polylactic
acid (PLA), and bio-polyethylene. Hybrid plastics are synthesized by
mixing algal biomass as filler material with petroleum derived plas-
tics. These materials have biodegradability properties and further
more decrease the quantity of petroleum derivative for plastic produc-
tion. The cellulose based plastics are mainly derived from cellulose re-
mains in algal biomass residue after extraction of algal oil. The
process is cost-effective, and algal biomass residue serves as ideal
feedstock for cellulose-based plastic production (Haniewicz et al.,
2018). Another form of bioplastics are PLA, which are produced from
polymerization of naturally produced lactic acid from fermentation of
natural feedstock. Algal biomass could serve as a better substrate for
lactic acid production via bacterial fermentation (Rhim, 2013). Ethyl-
ene monomer polymerizes to produce bio-polyethylene. The major
production of ethylene is carried out by cracking and dehydration of
ethanol, hence algal biomass could be used to produce ethanol as an
alternative substrate for bio-polyethylene production (Noreen et al.,
2016).
4.8. Nanocellulose
Compared to terrestrial plants, algae has gain popularity as a valu-
able source for nanocellulose production near future. Few reports are
available till date on successful extraction of nanocellulose fibers from
green filamentous algae such as Cladophora, Chaetomorpha, Microdyc-
tion, Rhizoclonium and members of Siphonocladales. (Liao et al., 2015).
Previously overlooked, however, the cellulose present in algal biomass
residue after oil extraction can be converted to produce nanocellulose,
an industrially important biopolymer having unique features such as,
renewability, environmentally benign nature along with its mechani-
cal properties. The main advantages of using algae for the biopolymer
production is very less content of lignin compared to the cell wall of
woody biomass, thus it involves mild chemical treatment for delignifi-
cation and reduced timing for high energy consuming mechanical
treatment procedures for yielding high degree of crystallinity in the
produced nanocellulosic material (Lee et al., 2017; Guo et al., 2017).
The market demand for nanocellulose has increased exponentially ow-
ing to its wide range of possible applications in various industries
such as food, pharmaceutical, agriculture, water treatment etc.
(Menon et al., 2017).
4.9. Algae derived biocomposites
Since past few years, there have been focused research interests on
production of microbial biodiesel which is significantly flourishing
due to capability of some microalgae to deliver high amounts of lipid
under controlled culture conditions (Chandra et al., 2019). Although
there is lack of studies regarding effective utilization of the microalgal
residues, which is left after lipid extraction but still enriched with sig-
nificant amount of proteins, carbohydrates or cellulose. Hence, it can
be used as an economic and environmental friendly filler alternative
for production of bioplastics. Recently, some researchers like Toro et
al. (2013) have reported production of bioplastics having optimized
composition of poly (butylene succinate) (PBS) and microalgal
residues after oil extraction in the range of 20–30%. Similar reports
M. Bhattacharya and S. Goswami
were demonstrated by Torres et al. (2015), who have used poly (buty-
lene adipate-co-terephthalate) (PBAT) and microalgal residuesin 4:1
ratio for preparation of biocomposites. Nonetheless, Tran et al. (2018)
have most recently reported about the improvement of mechanical
and thermal properties of prepared bioplastics fabricated with 20% of
Nannochloropsis salina residues along with PVA (Tran et al., 2018).
5. Bioprocessing of microalgae
Numerous studies have effectively used microalgae for bioproduct
generation. For a successful biorefinery primary phase include up-
stream and downstream processing among which, the productivity of
upstream processing includes four critical elements, to be specific:
strain of microalgae, carbon dioxide (CO2) supply, nutritional supple-
ments, for example, sources of phosphorus and nitrogen, and intensity
of light (Vanthoor-Koopmans et al., 2013). For microalgae culture de-
velopment, the sources of supplements are crucial as they give the vi-
tal parameters for culturing microalgae. Moreover, it was accounted
for that light intensity and the light source was more noteworthy as it
significantly affect the photosynthesis rate of microalgae hence the
development rate (Aravantinou and Manariotis, 2016).
Regular downstream processing unit forms that happen inside the
photobioreactor are the extraction and filtration strategies to get im-
portant mixes from microalgae. Ordinary techniques such as homoge-
nizers, bead beating, high pressure heating and synthetic compounds,
can acquire staggering expense of generation and the absence of a
conservative procedure has instigated the requirement for the combi-
nation of numerous means (Jacob-Lopes et al., 2015). The collecting
of biomass pursued by biorefinery systems has helped much in the
combination of biomass change forms. The utilization of mild tech-
niques of separation, which don't include solvents and high pressure,
are huge for the generation of the ideal compounds without harming
different parts (Vanthoor-Koopmans et al., 2013). Choice of proper
advancements for microalgae biorefineries relies upon the required vi-
tality input and the accessibility of recent innovation. The combina-
tioiin of upgrade technologies into downstream processing ought to
likewise enhance the procedures regarding financial, straightforward-
ness and simplicity of resulting preparing steps.
Be that as it may, when the products are focused for restorative
biotechnology reason, the development of microalgae ought to be
conducted under all around standardized conditions to maintain a
strategic distance from microbial tainting or the nearness of polluting
influences to meet the administrative necessity. From this viewpoint,
open development frameworks, for example, open lake or raceway
lake, may not be reasonable for developing microalgae that would be
utilized for restorative or pharmaceutical applications, regardless of
that they are all the more financially practical for substantial scale de-
velopment (Milano et al., 2016). Conversely, closed culture frame-
works (e.g., photobioreactors) with extensive control on the develop-
ment conditions (e.g., temperature, pH and CO2 focus etc) would be
increasingly plausible for this reason, though the capital and working
cost will increase (Matsui et al., 2019).
5.1. Algae culturing
Large-scale cultivation of microalgae can resolutely aid in the
emergence of renewable resource based industries for producing cost
efficient valuable products alongside biomass generation. Large scale
cultivation potential can be found from several species of microalgae
however there's scarcity of data for commercial trial runs. As such for
the case of bioethanol production, a large quantity of algal biomass is
needed to contend with alternative feedstocks like agricultural bio-
mass, thus for microalgae culturing technologies, it have to be com-
pelled to produce high yield of biomass, so that the application of
agricultural biomass for bioethanol production becomes relatively less
7
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
enticing. For algal culture to be feasible, the biomass production
should reach above 30 g/m2 a day (http://www.energy.gov/sites/
prod/files/2014/07/f17/mypp_july_2014.pdf.).
In various condition, different strategies can be used for cultiva-
tion of microalgae. With the help of light as associate energy supply
they convert water and environmental CO2 into biomass via biochemi-
cal process i. e. photosynthesis (Matsui et al., 2019). Synthesized
products are stored in numerous forms, like cell elements or storage
food materials, and vary from 20 to 50% of total biomass. Algae con-
jointly prefers nitrogen and phosphorus as major nutrients, that com-
prises of10–20% of alga biomass (Nagao et al., 2019). Alternative ne-
cessities for growth of microalgae includes the macronutrients in the
form of salts of Mg, Ca, and K; micronutrients, such as Mo, Mn, B, Co,
Fe, and Zn; and alternative trace elements. The preference of growth
media components varies according to different species. However, the
key necessities remain similar for nearly all species and embody es-
sential nutrients (N, P, K etc.), associate organic or inorganic carbon
supply, temperature (20–30 °C) pH (6–8) and salinity, oxygen, light
source (16 h light/8 h dark) and CO2 (Nagao et al., 2019). Waste wa-
ter from food and agricultural industries could also be a sensible sup-
ply of the specified nutrients for microalgae culturing technologies.
Another foremost vital factor for culturing algae is the usage of
different types of bioreactor. Bioreactor designing could be done fol-
lowing the species and also according to the product profile. On an
outsized scale, algae are cultivated in open ponds. This is well suited
for the species of algae that raise in heterotrophic culture (Morales-
Sánchez et al., 2017). Open culture systems are relatively cheap, how-
ever contaminate easily. Otherwise the options are to use continuous
or batch bioreactor facilities (Tardu et al., 2016). For industrial culti-
vation it's possible to cultivate waste water treatment plant with mi-
croalgae to urge twin benefits of remediation of waste water and pro-
duction of biomass. Marine water is additionally a decent variety for
industrial scale culturing of microalgae and ocean cultivation system
is gaining popularity among researchers and industrialists due to
some advantages such as natural mixing of culture by ocean waves,
exposure to dissolved nutrients on a large surface area etc. Exploita-
tion of sea water rather than pure water for culturing of microalgae
can cut back the value of production. Sea water could be a sensible
choice of media for microalgae cultivation to decrease the production
media costs and increasing product yield including lipids and alterna-
tive high value products in microalgae biomass (Park et al., 2018).
5.2. Harvesting
For open culture systems, harvesting microalgae from open culture
systems is associated with energy intensive and costly method of cen-
trifugation (Vandamme et al., 2013), which includes 20–25% of the
overall culture prices (Tsunemitsu et al., 2018), and thus different and
cheap harvest technologies are most well-liked research interests in
this field. For open culture systems (e.g., open ponds), flocculation is
taken into account to be a cheap harvesting technology for pre-
concentration of the culture (Vandamme et al., 2013). Inorganic floc-
culants, cationic polymers, bioflocculants, and additional recently, the
employment of nanoparticles as flocculants are some of the examples
used by several researchers (König et al., 2014; Roselet et al., 2015;
Alam et al., 2016; (Tsunemitsu et al., 2018), ‘t Lam et al., 2014). But,
the downside of flocculation technique is that the presence of floccu-
lant residues within the harvested algal cells also as within the
processed cell free medium, which can lower the worth of the har-
vested biomass and that of the extracted product significantly. Addi-
tionally, to resolve the issue, development of food grade or recyclable
flocculant is needed which will add extra cost to the culture price
(Eyley et al., 2015).
The second most important harvesting technology is microfiltra-
tion, which is a low energy consuming procedure and consumes en-
M. Bhattacharya and S. Goswami
ergy below 1 kWh/m3 as reported in recent literatures (Gerardo et al.,
2015; Ruiz et al., 2016). Bilad et al. (2012) have worked with micro-
filtration technology for pre-concentrating algal biomass from open
culture systems employing submerged membranes at an energy input
of solely 0.27–0.41 kWh/m3. While the step of pre-concentration is
necessary for the microalgae cultured in open ponds before to scale
back the prices of harvest microalgae, but it is not essential for har-
vesting microalgae if they were cultured in an additional targeted cell
suspension system bioreactor like PBR. In this case, for large scale cul-
tivation, centrifugation is the method of choice for harvesting mi-
croalgae, as the pre-concentration step is not required and will ac-
count for less than 5–7% of the overall cultivation prices as reported
within the techno-economic analysis reports by various researchers
(Ruiz et al., 2016; Chauton et al., 2015). This makes harvesting of mi-
croalgae much more preferrable from closed cultivation systems.
Microalgae harvest technology could be a newly developing field
with newer innovations incorporating filtration strategies projected
for harvest microalgae from open culture systems. At this point of
view, At recent techno-economic evaluations studies predict that har-
vesting algal biomass from closed PBRs is turning more economically
profittable. But, further analysis is required as a result of existing in-
formation on separate harvesting techniques are scanty. It's vital to
comprehend, however, that a selected harvest technology could be ap-
plied otherwise for various microalgal species irrespective of the ex-
pected specifications (type of PBR, species of microalge, etc.) however
conjointly biological aspects specific to strain (Vandamme et al.,
2016).
5.3. Cell disruption
Microalgal cell disruption could be a necessity for the entire biore-
finery as a result of the plasma membrane and membrane represent a
barrier to the discharge of intra-cellular compounds. There are three
techniques for cell disruption process of microalgae likewise biochem-
ical cell disruption (e.g., protein or alkali or heat treatment), mechani-
cal technique for cell disruption (e.g., bead beating and high pressure
homogenization), and physical process for cell disruption [i.e., Pulsed
electric field (PEF) or ultrasound treatment]. Cell disruption ought to
rather be performed beneath delicate conditions. These conditions are
imortant to take care of the original conformation of components of
individual cell (Günerken et al., 2015). Particularly, soluble proteins
are sensitive to high temperature and pH and ought to stay intact as a
result of they will have high price due to their techno-functional prop-
erties (e.g., emulsifying and foaming properties).
Mechanical processes don't need elevated temperatures or high
pH, and therefore they are a decent selection for delicate cell disrup-
tion (Günerken et al., 2015). Recent studies reported, that native algal
proteins were free when bead beating was done, however the energy
consumption of those processes would be high (Coons et al., 2014). In
contrary, some literature reports also reported that there is decrease
in consumption of energy by mechanical disruption processes signifi-
cantly by the systematic improvement of operational conditions. For
example, in studies held recently by Postma et al. (2017), reported re-
duced energy consumption during bead beating from 0.8 to 10 kWh/
kgDW to 0.5 kWh/kgDW by optimizing bead size and time of treat-
ment (i. e., energy input) for yielding the targeted proteins from algal
biomass. However, the major drawback of bead beating process is
that, the cells are completely disintegrated into very minute frag-
ments, which forms a stable emulsion having heterogeneous mixture
of cell components, from which separation and extraction of targeted
compound is a troublesome process (Günerken et al., 2016;
Montalescot et al., 2015).
Among physical cell disruption technologies, the PEF procedure
could be a low-shear and low-temperature technology and will en-
hance the extraction performance of hydrophobic components from
8
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
the biomass (Goettel et al., 2013; Luengo et al., 2014). For a whole
biorefinery that conjointly aims at getting practical and soluble pro-
teins, a dual stage approach was projected during which plasma mem-
brane is removed first then the inclined cell membranes are PEF-
treated (‘t Lam et al., 2017). However, due to the sturdy outer plasma
membrane, several microalgae don't seem to be at risk of PEF treat-
ment, consisting of the discharge of enormous intracellular com-
pounds (‘t Lam et al., 2017; Postma et al., 2016). Therefore, no yields
just like those deduced when mechanical cell disruption have been re-
ported, and also the overall method becomes additional advanced.
The selective removal of the plasma membrane barrier needs a more
robust comprehension of the chemical composition and also the com-
ponent of the plasma membrane of the microalgae (Gerken et al.,
2013; Scholz et al., 2014). Once the plasma membrane physiology is
known, one might easily design acceptable techniques of delicate cell
disruption.
5.4. Extraction
Extraction of different valuable components from microalgae are
typically done by organic solvent extraction, by which the hydropho-
bic lipids/pigments can be extracted, but the hydrophilic components
remains as residual solid fraction (Mubarak et al., 2015). Hence, it is
generally combined with a preliminary drying step to increase the ex-
traction yields of lipids (Du et al., 2015). However, this drying step
can be bypassed if amines or amidines (primary, secondary or ter-
tiary) were used as solvents which have inducible polarity i. e. these
solvents can switch reversibly to a polar or associate apolar solvent
when sparged with either CO2 or N2 (Du et al., 2015). This property
enables the solvents recovery (‘backextraction’), creating it a compre-
hensive technology for wet lipid extraction. In case of a complete
biorefinery, to valorize all the fractions of biomass, it is needed to ob-
tain alternatives for replacement of organic solvents because proteins
have tendency to denature throughout extraction with polar or am-
phiphilic organic solvents (Desai et al., 2016). It's convenient to use a
two stage extraction method during which removal of soluble proteins
occurs prior to the extraction of lipids from the biomass. Recently, a
new technique for the extraction process of soluble proteins was de-
veloping utilizing systems that are aqueous and biphasic. Future
analysis ought to concentrate on more characterization of those sys-
tems, permitting a additional rational style of delicate high value mol-
ecule extraction processes (Desai et al. 2014, 2016).
In recent times, another promising approach is now gaining inter-
ests for downstream processing eg microalgal biomass is to utilize sys-
tems supported with ionic liquids (ILs) (Orr et al., 2016). ILs are kind
of organic salts that are non-volatile and in liquid state at room tem-
perature. Their non-volatile nature will offer them benefits over vari-
ous organic solvents, but the problem arises if the extractable product
also have low vapour pressure at room temperature, as a result of this
may hamper back extraction (Desai et al., 2016; Orr et al., 2016).
Mostly ILs are imidazolium-based and employed with success for ex-
traction of lipids from wet algal biomass (Orr et al., 2016). Appar-
ently, IL based systems (Iolylite PG/sodium potassium phosphate)
may also be used for the hydrophilic native proteins extraction from
harvested biomass, as reported by Desai associated colleagues (Desai
et al., 2014). Although there are many convincing techniques for the
extraction of each fraction from biomass of microalgae, this approach
of single-component can be in demand for an exceedingly multi-
product biorefinery.
6. Product portfolio from current method
There are plenty of literature reports written on present under-
standing of microalgae derived products (Bharathiraja et al., 2015;
Koller et al., 2014; Lu et al., 2014; Bastiaens et al., 2017, Voort et al.,
M. Bhattacharya and S. Goswami
2015). Herein this section, we have tried to provide up-to date data
about the microalgae biorefinery practices for high value products
with respect to their future industrial applications. Information about
commercial production of microalgae are though scanty, but estima-
tion of approximate data of worldwide production volumes and their
prices for bulk cultivated biomass are roughly 10,000 tons/year and
$10,000/ton for Spirulina, 4000 tons/year and $20,000/ton for
Chlorella, 1000 tons/year and $20,000/ton for Dunaliella, and 200
tons/year and $100,000/ton for Haematococcus (Voort et al., 2015;
Benneman, 2013). Alternative species like Tetraselmis and Nan-
nochloropsis also are cultivated commercially, however the large scale
production in tons have not achieved yet.
Most of the algal biomass products are sold as whole dried powder
which contains a wide selection of claimed activities related to human
health such as enhanced immunity, preventing cardiac problems, obe-
sity and hypertension, and anticarcinogenic effects also (Spolaore et
al. 2016). However, human consumption of microalgal/cyanobacterial
biomass remains restricted to a really few taxa, as an example, Spir-
ulina, Chlorella, Dunaliella, Tetraselmis, Nostoc, and Aphanizomenon
(Spolaore et al. 2016; Voort et al., 2015; Kovac et al., 2013). The
combination of commercial food products with whole dried microal-
gal biomass still have not possible except only minor amounts (<1%),
as most of alga species tend to possess a strong green color, fishy
odor, and a powdery consistency (Kovac et al., 2013).
Majority of alga ingredients which represent high-value com-
pounds in the form of colorants/pigments and PUFAs, are presently
sold in markets for the food, pharmaceutical and (aqua-)feed sectors.
At present, the sole pigments on the market are β-carotene from
Dunaliella salina and astaxanthin from Haematococcus pluvialis
(Borowitzka, 2013; Voort et al., 2015). The extracted carotenoids are
employed in a range of applications, varied from being a source of
natural pigmentation supply for cultivation to food coloring functions
such as in chewing gums, dairy products jellies, etc. The other major
uses of pigments are in cosmetics like lipstick and eyeliners mostly in
Japan, Thailand, and China (Kumar et al., 2014). Alternative main
components embody lipids, which might be utilized in energy sector
as a supply for biofuels, in chemical industries as a supporting mater-
ial, in food industries as edible oil, in cosmetics and pharmaceutical
industries as antioxidants/glycolipids/phospholipids (Draaisma et al.,
2013; Cuellar-Bermudez et al., 2015; Voort et al., 2015; Leu et al.,
2014). The other component carbohydrates are commonly consist a
minor portion which needs to be explored from a couple of algae
species (García-Martínez et al., 2016). The advanced (excreted) poly-
saccharides may need health useful properties, a motivating niche
within the healthcare sector, whereas the carbohydrates within the
cell (e.g., starch and hemicellulose) is used for manufacturing
bioethanol or bioplastics and chemicals like nanocellulose (García-
Martínez et al., 2016).
The overwhelming yearly alga biomass production (Arthrospira
and Chlorella) is presently depended on the whole dried biomass to
produce powders, pills, and tablets (García-Martínez et al., 2016.
Hence, majority of alga process solely contains a prior harvesting step
and an ulterior drying step, mostly done by spray drying process as
per the commercial websites of earthrise (http://earthrise.com/about/
our-farm/), cyanotech (http://www.cyanotech.com/company/
process05.html), and sunchlorella (http://www.sunchlorella.com/
corporate-activity/manufacturing-process.html)). Alternative products
on the market are pigments/carotenoids like β-carotene and astaxan-
thin, that are made from Dunaliella salina and Haematococcus pluvialis,
severally. The extraction steps are mostly remains same except for
Dunaliella that lacks a robust plasma membrane, bypassing the re-
quirement for cell-disruption, however for Haematococcus, which con-
tains a sporopollenin plasma membrane, making it compulsory to ap-
ply a cell disruption step before the astaxanthin extraction protocol.
9
Biocatalysis and Agricultural Biotechnology 25 (2020) 101580
For pilot scale production, automatic cell disruption was done with
bead edge or expeller pressing.
Hydrophobic components extraction (carotenoids or lipids) are
done through exploitation of organic and sub- or supercritical sol-
vents. For industrial scale production, there's a niche for supercritical
carbon dioxide gas extraction technology, which offers the very best
yield for antioxidant recovery, however, it has the disadvantage of re-
quirement of that it's operating high pressures and energy that the
method needs. But, for high value applications like nutraceutical
product development, this technology appears as the foremost eco-
nomical answer. However, the measurability apart from the nutraceu-
tical market appears rather restricted. Shrimp feeding trials are done
with the help of defatted Haematoccocus pluvialis genus to interchange
part fish meals in cultivation, that failed to have severe effects on
growth performance and nutritionary composition, and gave vital pig-
mentation of the shrimp. Many approaches are done to purify C-
phycocyanin, betting on the required purity (Minkova, 2013). It al-
ways involves a mixture of ammonium ion salt precipitation, organic
solvent extraction process, and gel filtration natural process (based on
the specified purity). No literature may be obtained on the pertinence
of the remaining biomass, though it's still made in lipids (e.g., γ-
linolenic acid), proteins (membrane proteins), and carbohydrates.
7. Industries involved in algal technologies
Currently there are many researchers, research organizations, in-
stitutes and companies, working throughout the world to obtain mi-
croalgal biofuel and other value-added products in a more economical
and sustainable way. However, there are few companies working on
microalgal biorefineries which are established and have a remarkable
presence in global market (Ritala et al., 2017; Chandra et al., 2019).
To name a few, Astaxanthin pigment which is used as biopharmaceu-
tical is being produced from algal source and marketed by Algatech-
mologies, BioReal Inc. and Cyanotech Corporation. Astaxanthin from
Haematococcus pluvialis has also been produced by a marine biotech-
nology company Mera Pharmaceuticals Inc. Cyanotech Corporation
has been known as world’s leading producer of Spirulina as food in-
gredient which is FDA approved and has been sold in more than 30
countries worldwide including US. Proteus Corporation which is
founded in 1976 and progenitor of Earthrise, are also second largest
producer of Spirulina with GRAS status having GMP and Food Safety
System Certification (FSSC). Parry Nutraceuticals Division from E.I.D
Parry Ltd. is part of Murugappa Group, also involves in microalgal
production of Spirulina and Chlorella as nutraceuticals products,
which are sold in more than 40 countries including North America,
Europe, South East Asia, and the Far East. In USA, Algeon Inc. has
been founded in 2011 and established as renowned producer Spir-
ulina and Chlorella. Their products are mainly based on b-glucan and
whole cell products from the photosynthetic protist Euglena gracillis.
Amongst other companies, TerraViaHoldings, Inc. is a publicly held
American company which focuses on providing food ingredients from
eukaryotic algae. In Germany also, there is a leading microalgal
biotechnology company, BlueBioTech Int. GmbH, which has operated
for more than 10 years, producing large quantities of Spirulina and
Chlorella. In Japan, Euglena Co. Ltd. Founded in 2005, is developing
de-fatted Euglena as a source of protein-rich animal feed derived from
Euglena gracillis. Table 2 summarizes various companies which are
working from identifying and optimizing specific strains of algae to
develop a sustainable biorefinery involved in the production of biofu-
els, protein feedstock, and nutraceuticals.
8. Opportunities and constratints of microalgae biorefinery
In spite of the good promise observed by the algal biorefinery, the
present drawbacks have to be compelled to be addressed. The overall
M. Bhattacharya and S. Goswami Biocatalysis and Agricultural Biotechnology 25 (2020) 101580

Table 2 be reviewed here intimately. Secondly, the biorefinery structure relies

Worldwide companies involve in microalgae research, their products and type on the ratio of the cell composition. Composition of algal cell isn't a
of cultivation method (Adapted from Chandra et al., 2019). set property, however relies on the strain also because the culture
conditions, specifically nutrients, temperature and, light (Juneja et al.,
Company Product Cultivation system References
2013). So ratio is tailored with regard to process options like energy
Algatechnologies Astaxanthin Closed and semi- Panis and requirement and productivity on the one hand and to promote de-
(Israel) closed bioreactors Carreon mands on the opposite. In alternative divisions of chemical engineer-
under high light (2016)
intensity
ing this seems to be a drag for several stoichiometrically coupled co-
BioReal Inc (USA) Astaxanthin Indoor Shah et al. products. Glycerine formation from oilseed primarily based biodiesel
photobioreactor (2016) production could also be given here as associate example. Typically,
Cyanotech Astaxanthin, Spirulina Raceway pond and Panis and shifting microalgal metabolism in favor of the formation of a high-
(Hawaii) pacifica as food ingredient photobioreactors Carreon
value product, e.g. by high light intensity, leads so to the next produc-
(2016)
Mera Astaxanthin from Raceway pond Brennan
tivity however at the expense of energetic potency of the whole
Pharmaceuticals Haematococcus pluvialis and method. This is often as a result of illumination within the saturation
Incorporation Owende vary doesn't cause the next energy turnover at the cellular level, how-
(2010) ever only promotes product formation. An extended light exposure so
Algenol (USA) Bioethanol, and pigments Raceway pond, ElMekawy
ends up in additional product however leads to reduction of overall
Closed and semi- et al.
closed bioreactors (2016) potency of energy.
Sapphire Energy Biofuels and Uses non- potable Maity It is not solely the distribution of various high value products for
Inc. (USA) water, seawater (2015) various markets that must be thought of. The educts like nutrients,
Solazyme Inc. Biodiesel and biojet fuel Heterotrophic Menetrez water, carbon dioxide gas and light ought to be employed in clever
(USA) cultivation and (2012)
ways in which to form the biorefinery successful. This suggests that as
photobioreactor
Sea6 Energy Food Additives, biofuel, Sea water Wei et al. several energy flows and material side-streams as attainable ought to
(India) bioplastic, animal feed (2013) be closed the plant itself. The requirement for closed material and en-
Muradel Pty Ltd. Biofuels, oleochemicals, Raceway pond Duong et ergy cycles offers a third side in an exceedingly broader sense of
(Australia) biofertilizers, animal feed al. (2012) biorefinery.
and building materials
Cellana (USA) PUFAs, animal feed, Open- pond bioreactor Menetrez
biodiesel and bio jet fuel (2012) 9. Techno-economic analysis and life cycle assessment (LCA)
Solix Algadrients Astaxanthin and DHA Enclosed Radakovits
Inc. (USA) photobioreactors et al. Microalgae have shown several features like yielding high lipid
(2012) content, possibility to grow in arid lands and water, and utilization of
carbon di-oxide sources. These characteristics attracts attention
amount of microalgal biomass made currently in industrial processes among researchers in view of the enhancements of the process steps
is substantially low (around 15000 tons per year) and processes of for the assembly of microalgae, that might produce high value prod-
production are sometimes economically not viable. High-value prod- ucts and biofuels. Currently, studies are centered on the progression
ucts from microalgae are sometimes considerably costlier than their and upscaling of the process and to find industrial feasibleness of fuel
chemical counterparts and serve niche biomarkets. Bulk products like production through microalgae-based feedstock. The techno-economic
biofuels are situated at the lower finish of the price scale, although assessment (TEA) and life cycle assessment (LCA) are typically uti-
biofuels should be comparable with low cost fossil fuels, but have to lized fundamental analysis tools for examining the feasibleness of mi-
be compelled to need a lot of less auxiliary energy that is finally cer- croalgae biofuels and other high value products. These tools enable to
tain within the biomass. This demand has not been satisfactorily analyze the choice of process technologies and to design production
achieved. Within the field of medium-value products for feed, e.g. in process that concentrate on the purpose and analysis efforts to realize
cultivation, food or for chemical building blocks, a breakthrough industrial usability (Quinn and Davis, 2015).
could also be on the horizon. Technologies for algae cultivation like open pool system (OP), and
How will the concept of the biorefinery facilitate to beat the pre- photobioreactors (flat panel, vertical and horizontal) are applied and
sent economic limitations? The fundamental concept is to assemble evaluated for the industrial feasibility of the systems of biorefinery.
high- or medium-value product with the use of the residual biomass. For a successful biorefinery system, with great economic and environ-
Compared to plants, microalgae haven't any roots or stems and lack mental performances ought to utilize residual biomass as animal feed
extremely resistant plasma membrane elements like lignocelluloses, substitute and the lipids for generation of vegetable oils severally. On
that are terribly troublesome to degrade and to with chemicals exploit the other hand, volumes of petroleum derived energy consumption
(Zeng et al., 2014). This greatly simplifies the combination of the use and emission of greenhouse gases are observed in such scenarios
of products from algal and residual biomass. The foremost obvious wherever the oil-free residue is anaerobically digestible to produce
possibility is to use it dynamically, from algae is a wonderful substrate biogas and also the lipids are regenerated to biodiesel (Posada et al.,
for the biogas fermentation (Wang et al., 2013) or thermal conversion 2016). In contrast with photobioreactors (PBR), OPs contains less
processes (López Barreiro et al., 2013). Alternatively, it's attainable to complicated construction and operations, resulting in lower capital
extract and sell secondary products or simply dry the residual biomass and operation prices. But, the advantages of using PBRs are more in
as a whole for animal feed functions. terms of productivity of microalgae, potency of uptake of nutrient,
Other possibility is recycling of the cellular components at inter- culture density and controllability.
vals between the process. as an example, nucleic acids from residual Thomassen et al. (2016) performed a techno-economic assessment
biomass would supply source of nitrogen to come up with contempo- steered that employing a specialised filtration membrane for recycling
rary, product forming biomass. To that extent, drawbacks of the mi- of medium and open pool cultivation of alga were the foremost
croalgal production processes that have to overpower depends on techno economically feasible and profitable possibility. PBR integra-
many basic aspects. Firstly, technical suggests that for acceptable sep- tion to the system was observed to extend capital investment price in
aration, extraction routes and conversion routes ought to be devel- spite lowering the necessity of medium (Thomassen et al., 2016). The
oped and are presently beneath investigation. The state-of the-art process of land occupation conjointly remains a confrontation as envi-
technology (Grima et al., 2013; Bahadar and Bilal Khan, 2013) won't

10
M. Bhattacharya and S. Goswami Biocatalysis and Agricultural Biotechnology 25 (2020) 101580

ronmental impacts have to be compelled to be taken into account for Table 3

land use. Consumption of excessive water also will cause water Some recent patents on microalgae biorefinery and related products.
scarceness which can have an effect on the water prices, and addition
Patent number Publication Assignee/Inventor Title
of water treatment method might needed to get ambient water quality date (s)
for applications in food. What is more, the uncertainties related to the
process scale-up of the emerging algal biofuel business produce an ab- US10119947B2 2018- 11- 06 Corbion Biotech, Inc. Protein- rich microalgal
biomass compositions of
sence of public agreement in light of the short term economical viabil- optimized sensory quality
ity of algal biofuels. US10098371B2 2018- 10- 16 Solazyme Roquette Microalgal flour
The major factor for high production cost of microalgae is the Nutritionals, LLC
process for growing and harvesting the algal biomass. The growth WO2017033188A1 2017- 03- 02 Yeda Research And Algal oil and biofuel and
Development Co. methods of producing
condition of algae, whether in open pond or in the artificial culture
Ltd. same
condition, also determines the total production content. During anaer- US9719114B2 2017- 08- 01 Terravia Holdings, Tailored oils
obic digestion of feedstock biomass, the growth of algae depends Inc.
upon the amount of water since nutrient are recycled during anaero- CN105861379 (A) 2016- 08- 17 Jiangnan U.; Yang Method for preparing
bic digestion and carbon-di-oxide is freely accessible in the atmos- H., Ping S., Wang T., algae powder by drying
Wang W., Zhang L. peculiar smell- free blue
phere. Despite the many advanced technologies, the efficiency of bio-
algae
fuel produced from microalgae is much lower than the fossil fuel de- WO2016092583 2016- 06- 16 Lavanga V.; Lavanga Method for growing
rived products (Manganaro et al., 2015). However, lipid production (A2) V. and Farne S. microalgae, and device
from microalgae had shown many improvements, such that microal- for implementing said
gae with high lipid content can be utilized for production (Eppink et method
CN105483014 (A) 2016- 04- 13 Qingdao Kehai Production technology
al., 2017). Furthermore, with the increasing cost of oil production and
Biological Co., Ltd.; for high- density culture
its subsequent consumption lead to the increase in the utilization of of Chlorella by utilizing
microalgae derived products for energy production. fermentation method
Life cycle assessments (LCA) for the analysis of impact of produc- US2016021923 2016- 01- 28 Solazyme Inc.; High- Protein Gelled Food
(A1) Paulsen S., Products Made Using
tion of algal biomass on environment is also very crucial. LCA studies
Klamczynska B., High- Protein Microalgae
analyze the impact of substrate, methods, by-products produced dur- Plasse K.,
ing biomass production, types of microalgal strains for particular Bowman C.
products and various other impact on environment production US9249252B2 2016- 02- 02 Solazyme, Inc. Low polyunsaturated
(Adesanya et al., 2014). These abstractions promote the use of mi- fatty acid oils and uses
thereof
croalgae in a suitable manner. Many studies have revealed that, re-
US2015374012 2015- 12- 31 Solazyme Inc.; High- Protein Food
covery of lipid combined with anaerobic digestion produces gas. The (A1) Klamczynska B., Products Made Using
combustion of these gases are used for power and heat generation. Echaniz A., Zhu R. High- Protein
Moreover, the analysis of environmental impact of microalgae de- Microalgae
pends upon the algal strains and the lipid concentration of the algal WO 2013086302 2013- 06- 13 Old Dominion U. Fractionation of proteins
(A1) Research and lipids from
cells.
Foundation; Kumar, microalgae
The LCA study for heat generation from anaerobic digestion had S., Hatcher, P. G.
shown that, generated heat is quite low as compared with the photo- US8187860B2 2012- 05- 29 Solazyme, Inc. Recombinant microalgae
bioreactors (Quinn et al., 2014). To boost the utilization of microal- cells producing novel oils
gae, as substrate for energy production, various additional modifica- WO2012038978A1 2012- 03- 29 Council Of Scientific Integrated process for the
& Industrial production of oil bearing
tions are needed to be done. A LCA study by Soh et al. (2014), found Research (An Indian chlorella variabilis for
three main factors affecting the environment-need for energy, emis- Registered Body lipid extraction utilizing
sion of greenhouse gases and presence of dissolved oxygen and min- Incorporated Under by- products of jatropha
eral content in the water. The study was done by screening numerous The Registration Of methyl ester (jme)
Societies Act production
algae from fresh and saline water, and growing them under different
US7977076B2 2011- 07- 12 Genifuel Corporation Integrated processes and
conditions of growth. It was observed that, increasing the algal bio- systems for production of
mass production did not shown a good impact on the environment. biofuels using algae
Out of the three environmental conditions mentioned above, only the US 20100303990 2010- 12- 02 Solazyme, Inc.; High Protein and High
presence of nutrients served to enhance the biomass content without (A1) Fiber Algal Food
Materials
showing any deleterious impact on the environment.
WO2010080377A1 2010- 07- 15 Alpha- J Research Optimization of algal
Be that as it may, the impacts on life cycle for cultivation of mi- Limited Partnership product production
croalgae are perceptive to parameters like availableness of renewable through uncoupling cell
sources and also the public demand for products based on microalgae. proliferation and algal
These framework might affect the LCA of the cycle of microalgae product production
process to biofuel and co-products. More analyses are expected to be
performed on the contents of lipid, production at stabile state and cli- component in feed mixture, which can be a good protein or nutrient
mate suitableness of sources of microalgal biomass to completely ap- substitute for farmed animals feed or fish feed. In recent years, though
praise the LCA. microalgae research has been conducted worldwide, but industries
and universities in China have been found most active in filing
10. Patent trends related to microalgae biorefinery products patents related to microalga biorefinery following USA. In current sce-
nario, there has been a surge to use microalgae as food supplement to
Microalgae biorefinery related patents filed in past 10 years has address food security problems worldwide. In this regard, production
been listed in Table 3. The filed patents are evenly distributed regard- of single cell protein have been reported by Ritala et al. (2017) by fer-
ing utilization of microalgae for biofuel production and other value- mentation of agricultural or food residues using mied population of
added products as reported by Talebi et al. (2018). There are also microalgae, bacteria and yeast. Thus, microalgae biorefinery can be
patents which discloses that microalgae biomass can be used as a simultaneously performed with waste bioremediation also. However,

11
M. Bhattacharya and S. Goswami
biofuels from microalgae are the most common attribute to generate
patents, but generation of value-added products along with continued
developments in microalgae cultivation methods are also attracting
attention following recent patent trends.
11. Conclusions
The overall production summary of commercialized products de-
rived from microalgae considers certain factors including the type of
algal strains, lipid composition of the cell, strategies for product for-
mation, ease of the overall process, use of substrate to carry out reac-
tions, amount of products and by-products formed and finally the im-
pact of overall process on the environment. The isolation strategies
from cells of algae again depend upon the way to disrupt cells, nature
of molecules-hydrophobic or hydrophilic, concentration and product
purification steps and its costs. However, the production is not so
easy, it seems that the factors like, high production costs, contamina-
tion by other microbes, low productivity, difficulty in down-streaming
process and many other factors limits its production. The biorefinery
of microalgae system for the assembly of biofuels and plenty of per-
ishable products, the ultimate produce are sometimes low because the
range of steps needed to obtain defined purity extent varies for every
industry, although method integration is accustomed to cut back the
amount of purification steps. Many issues that require to be addressed
are for comparatively small marketplace for totally varying products,
losses resulting from degradation of product, conditions of culture
growth and their effects also as long term stability studies of the algal
products. These studies are vital as compounds like pigments will sim-
ply detoriate because of light, temperature, and alternative microor-
ganisms. Lastly, development of a suitable method for easy and
cheaper production along with comprehensive studies of economic
and environmental conditions have to be compelled to conduct, con-
cerning the assembly of high-value products from microalgae, addi-
tionally to it, analysis of life cycle compounds of high-value from mi-
croalgae can be performed for the evaluation of the sustainability of
the procedure. Therefore, additional efforts ought to be performed to
scale back loss of product, minimize energy prices associated whereas
leading through a sustainable large scale downstream process for high
value compound extraction from microalgae and its subsequent con-
sumption declining the dependence on petroleum derived products.
Ethical approval
This article does not contain any studies with human participants
or animals or any microbial strains performed by any of the authors.
Declaration of competing interest
The authors declare they have no conflict of interest.
Acknowledgement
The authors want to acknowledge Center of Innovative and Ap-
plied Bioprocessing, Mohali, to carry out the review work. The au-
thors do not receive any funding for this work.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.bcab.2020.101580.
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