Environmental Chemistry Letters

https://doi.org/10.1007/s10311-020-00999-7

REVIEW

Biofuels, biodiesel and biohydrogen production using bioprocesses.

A review
Rajesh K. Srivastava1 · Nagaraj P. Shetti2 · Kakarla Raghava Reddy3 · Tejraj M. Aminabhavi4

Received: 6 January 2020 / Accepted: 24 March 2020

© Springer Nature Switzerland AG 2020

Abstract
Energy demands, pollution and global warming induced by globalization are rising, thus calling for alternative sources of
energies. In particular, biofuels are increasingly used for transportation, electric power and heat energy generation. Biofuels
can mitigate greenhouse gas emissions by up to 50%. Biofuels are produced from organic matter and waste such as dry
lignocellulose, algae, yeast, restaurant greases, food grain, non-food grain and animal fats. Biofuel from crop residues can
be promoted by government subsidies to reduce the fuel price and meet the requirement of industries, transportation and
agricultural sectors.

Keywords  Green environment · Biofuels · Organic waste · Fossil fuels · Environmental pollutants · Energy utilization

Abbreviations DOE Department of Energy

ACP Acyl carrier protein ER Endoplasmic reticulum
BTL Biomass to liquid EtOH Bioethanol
BTUs British thermal units eq/M.J Equivalent per megajoule
Cas CRISPR associated 2-EH 2-Ethylhexanol
CCS Carbon capture and storage FAs Fatty acids
CHP Carbon heat and power FT Fischer–Tropsch
CO2 Carbon dioxide g/g Gram by gram
CSLF Cellulose solvent-based lignocellulose Gt Gigatonnes
fractionation GHGs Greenhouse gases
CRISPR Clustered regularly interspaced short palin- H2 Biohydrogen
dromic repeats H2O Water
DME Dimethyl ether H2S Hydrogen sulfide
IBPGM Integrated biomass pyrolysis, gasification
and methanol synthesis
* Nagaraj P. Shetti KOH Potassium hydroxide
dr.npshetti@gmail.com LHV Lower heating value
* Kakarla Raghava Reddy MEC Microbial electrolysis cell
reddy.chem@gmail.com MeOH Methanol
1
Department of Biotechnology, GIT, Gandhi Institute MEF Microbial electrolysis cell
of Technology and Management (GITAM) (Deemed MFC Microbial fuel cell
to be University), Rushikonda, Visakhapatnam, Mo/Pt/WOx Molybdenum/platinum/tungsten oxide
Andhra Pradesh 530045, India MT Million tonnes
2
Department of Chemistry, Center for Electrochemical Mteo Million tonnes of oil equivalent
Science and Materials, K. L. E. Institute of Technology, NaOH Sodium hydroxide
Gokul, Hubballi, Karnataka 580030, India
NER Net energy ratio
3
School of Chemical and Biomolecular Engineering, The NOX Nitrogen oxides
University of Sydney, Sydney, NSW 2006, Australia
NMB National mission on biodiesel
4
Pharmaceutical Engineering, SET’s College of Pharmacy, OPEFB Oil palm empty fruit bunch
Dharwad, Karnataka 580 002, India

13
Vol.:(0123456789)

PAHs Polycyclic aromatic hydrocarbons
PBRs Photo-bioreactors
PEM Polymer electrolyte membrane
PS Photo-system
PoGA15A Starch-digesting glucoamylase
S. cerevisiae 
Saccharomyces cerevisiae
SNG Synthetic natural gas
SPSC01 Self-flocculating yeast strain
SO2 Sulfur dioxide
SO3 Sulfur trioxide
SSF Separate hydrolysis and fermentation
TAG​ Triacylglycerol
w/v Weight by volume
US United States
USA United States of America
XUSE Xylose-utilizing engineered S. cerevisiae
strain
v/v Volume by volume
Introduction
The development of renewable energy technologies for sus-
tainable biofuel synthesis can help us to provide a maximum
amount of our energy demands. Biofuel can operate in a
sustainable nature with more cost-competitive to existing
energy options such as fossil fuel. Increased fossil fuel util-
ity is caused by the population growth of many countries
for heating, lighting and transportation purposes due to
expanding the economy. Biofuel contribution can complete
the demand of 6% of total energy needs by about 2030 as
reported by the International Energy Agency expectation
(Shi et al. 2018; Kumari and Singh 2019).
Algal biofuel can be another best option to meet energy
needs, but it requires to solve many challenges such as
identification of and improvement in algal strain with more
quantity of oil contents and also crop protection, nutrient
resources, and its allocation as well as the production of
co-products with improved economy of the circular system.
It has many challenges for the economic scale-up of algal
biofuel and needs for improvement in the scientific commu-
nity to solve these challenges from promises to reality (Shi
et al. 2018; Yew et al. 2019; Yu et al. 2017).
Microalgal species with single-cell nature has shown its
potential for the development of liquid transportation fuels,
and these have shown the capacities for growth in marine or
freshwater resources with good carbon fixation capability for
the storage of high amount of oil contents in their biomasses
in bubble column photo-bioreactors that can be utilized for
the production of algal biodiesel development. Microalgal
species Botryococcus sp. has shown to store long-chain
hydrocarbon (50%) in their dry biomass, and other species
of algae have been identified to create ample opportunities
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Environmental Chemistry Letters
to discover good techniques (Kojima and Zhang 1999; Yu
et al. 2017).
Biodiesel is produced from vegetable oil, soy oil, canola
oils, cooking oils and animal fats. These oils are technically
known as acid methyl esters. Also, soy oil is the main or
common feedstock of the USA, and it is used to synthesize
biodiesel then known as soy methyl ester. Vegetable oil is a
triglyceride, which is utilized in the presence of a catalyst
under processes of transesterification for the synthesis of
diesel. Transesterification can break off glycerin component
via attaching each fatty acid to alcohol. This process starts
with the first step for breaking ester–glycerin bonds, and
NaOH and KOH have been added as the catalyst compo-
nents. It was found that this reaction is normally performed
in the presence of alcohols and free fatty acids. Biodiesel
has a chemically different nature compared to pure vegetable
oils via providing a substitute for diesel fuel (Rabin 2003;
Sahar et al. 2018). Producing sustainable transport nature
of biofuels from the microalgal species can help to develop
economic viability with eco-friendly sustainability via main-
taining the energy and carbon balancing with the environ-
mental impacts and reduced production prices. Microalgal
biofuel can be the basis for public support in the direction
of microalgal species or strain investigation in the current
research fields (Slade and Bauen 2013). In this review, we
will attempt to discuss biofuels production through various
bioprocesses and renewable resources for sustainable biofuel
development with their utility in different sectors worldwide.
Biofuels resources
Biofuels are synthesized from different types of biological
processes via the utilization of organic matters in living plant
sources, waste organic matters from plant or animal waste
residues. This energy fuel has no carbon emission in the
environment compared to petroleum or coal fuel burning to
meet our energy needs. It has helped us to achieve positive
energy balance that can be utilized by applying advanced
technical devices or information with the highly optimized
production, which can be more suitable for algal biofuel
development (Slade and Bauen 2013). Further, fertilizers as
well as energy requirements are used for drying and dewa-
tering. Limited sources of primary data for bioprocess and
scale-up of algal biofuel development are needed to advance
with highlighting of future uncertainties. Environmental
impacts of algal biofuel production are shown to manage
for safe handling of water resources and carbon dioxide as
well as nutrient supply as the major constraints of system
design and implementation options. The C ­ O2, nutrient and
water resources achieved at low cost can help in cost reduc-
tion in algal biofuel production by about 50% (Slade and
Environmental Chemistry Letters
Bauen 2013; Rawat et al. 2011; Barreiro et al. 2013; Roux
et al. 2017).
The net energy ratio (NER) can be applied for algal bio-
mass production by providing the idea of the total amount
of energy utilization for purposes of cultivation, harvesting
and drying processing with the division by the energy con-
tents of the dry biomass. Positive energy balance for algal
biofuel can thus be achieved from the utilization of culturing
systems such as raceway pond systems or photo-bioreactors
(PBRs) (Mutter 2019). Raceway pond systems are reported
as a closed-loop oval channel with a size of 0.25–0.40 m
deep, exposed to air with mixing facility (via paddle wheels)
for the circulation of water and prevention of sedimentation.
In the PBRs culturing system, media are found to enclose in
a transparent array of tubes or plates and broth of microalga
is found to circulate from a central reservoir. PBRs systems
can provide better control or regulation of the environment
of algal culture, but it is found to be more expensive than
the raceway culturing pond and it requires a high demand
for auxiliary energy (Mutter 2019; Yao et al. 2019; Chisti
2007; Narala et al. 2016).
South Korea has utilized the biofuel oils for the genera-
tion of electric power for their energy needs in various sec-
tors. Its ministry of trade, industry and energy has revised
the laws from January 2019 onward to utilize biofuel oil as a
source for cleaner energy production such as electric power
generation, which started in March 2019. From the biofuel
utility for electric power generation, South Korea has now
developed the capability for supplying electric power with
coal or nuclear power (non-renewable energy source). These
efforts can provide fuel oil power for small portions of this
country’s electricity needs (Tylor et al. 2016; Brigljević et al.
2019, Arifin et al. 2014). It was reported that utilization of
monster 250-m-long Whitechapel fatberg via conversion into
a more quantity of biodiesel (approximately 10,000 L) and
used for the utilization of congealed masses of fat, oil or
grease wastes (Water 2017).
The produced biodiesel was utilized as an eco-friendly
energy resource to power 350 buses of double-decker of
Routemaster. A third of the 130-tonne fatberg has hit the
headlines around the world, and these efforts were made
to remove them from the bowels of Whitechapel Road.
Whitechapel fatberg has shown its deserving capability for
waste conversion to power firm Argent Energy via trans-
forming into evil, gut wrenching and rancid blob into pure
green biofuels. Thames River Water was shown to be one
of the biggest producer companies for green energy inside
the M25 design, and they are continuously working for self-
generation of 33% of the electricity as renewable energy
sources by the year 2020. This company has reported for the
production of enough electricity to power more than 86,000
homes (Water 2017; Denton 2019). In this review, we will
also consider various bioprocesses and renewable resources
for sustainable biofuel development with their utility in dif-
ferent sectors worldwide.
Bioprocesses for the development of biofuels
Biofuels are available in solid, liquid or gaseous forms that
are biosynthesized for the utilization of organic matter from
fresh resources such as food grains, algal biomasses or non-
food materials as well as waste organic materials such as
agricultural residue wastes, milk–whey–sugar, wastewater
or any decaying organic materials as shown in Figs. 1 and
2. Biomasses as renewable resources can be utilized for the
biosynthesis of liquid biofuels with their applications in
transportation sector for their energy need to run the auto-
mobile engine (Pérez-Camacho et al. 2019; Baldino et al.
2018).
Common transport vehicles such as passenger vehicles,
trucks, ships and airplanes are dependent on gasoline, die-
sel or kerosene in their internal combustion engines, but
many efforts have been made to utilize sustainable biofuels
in the form of conventional or advanced technologies for
the production of ethanol and biodiesel. From the biofuel
utility, 80% of emission reductions in greenhouse gases can
be achieved compared to the fossil-fuel-powered engines.
Another utility of advanced biofuels can be as fuel require-
ment for heavy freight, shipping and aviation. Thus, there is
a need for the world organization to promote the commer-
cialization of advanced biofuels technology with enhanced
investments in biofuel production capacity. These efforts
may help to enhance the production of advanced biofuels
for energy requirements (Baldino et al. 2018; Moriarty et al.
2019).
Biofuels production is reported via the utilization of
heterologous aerobic nature pathways in yeast or bacte-
rial strain. It is required to control the aeration or cooling
processes at a large scale, and utilization of lignocellulosic
biomass has been reported via converting into fermenting
sugars. Biofuels synthesis requires the need for an effec-
tive biomass pretreatment for increasing the surface area or
decrystallized cellulose contents or components via facilitat-
ing enzymatic hydrolysis (Banja et al. 2019). Recovering of
microalgal strains and extraction of their intracellular lipids
have been reported as effective tools as an economic bottle-
neck in the area of algal biodiesel production. Cost-effective
deconstruction of biomass can thus be enhanced for the bio-
fuels production at a lesser cost. At any rate, algal biodiesel
production has some challenges such as cell harvesting and
lysis processing (Banja et al. 2019; Zhang et al. 2017).
By using bioprocesses, biomass conversion for the bio-
energy synthesis, Brazil agro-industry has reported their
great potentials as the feedstocks and raw materials utili-
zation. This bioprocess is especially used for the second-
or advanced-generation approach for ethanol and various
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Fig. 1  Agricultural waste utilization for sustainable bioenergy with
reduced emissions. This figure is organised in different steps. The first
is shown with solar energy, the second is green plants captured this
energy for their products (e.g., edible or non-edible nature), the third
explains the pretreatment process to partial loosen their plant product
for carbon source, the fourth shows loosed non-edible plant parts or
chemical syntheses. Earlier utilization of sugarcane juice
or its byproducts was done with the help of conventional
technology for the first-generation ethanol or other biofuel
production, but it has achieved around 23.64 billion liter
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Environmental Chemistry Letters
matter, the fifth shows the biological treatment to complete hydroly-
sis of plant matter, the sixth shows fermentable sugars, the seventh
shows microbial strain for fermentation, the eighth shows synthesized
biofuels, the ninth shows internal combustion, and the final or tenth
part shows least smoke formation from biofuel combustion
ethanol during 2012–2013 in Brazil (Gargalo et al. 2016;
Pfaffingera et al. 2019).
The recent advances in technological developments and
research for the most competitive ethanol production in the
Environmental Chemistry Letters

Fig. 2  Bioprocesses for biofuel
development. Here, the first hν~Solar Green plants Plants products Biofuel-processing step
section presents the use of solar energy Lipid content
energy by plants. The second
extraction and
section shows the first products
trans-esterification
of photosynthesis. The third
sector explains the details of
pretreatment processes for plant Microalgal species
product hydrolysis. The fourth
section gives examples of biofu-
els, and the fifth section shows Sun Pretreatment and
the utilization sectors
fermentation

Food grains

Biofuel
Agricultural wastes
Biofuels
Biofuel Utility
Transportation Ethanol
Green diesel
Energy Generation Biodiesel
Providing of Heat Biofuel Biogas
Wood, dried
Charging Electronics Utility
plant material,
Cooking
Lubricate
Removing paint and adhesive

hν~Solar
energy
1

1-Transportation
2. Charging electronics
3-Energy generation Algal 2.2a
4-Cooking Biofuel plants
5 Microalgal
5-Lubricating task utility 2.2 species
6-Removing
Biohydrogen
paint Biodiesel
Green 2.1:Higher
diesel plants
Ethanol 4
Butanol Biofuels Biogas
2.1a: Food
grains
Microbial fermentation
3.1-Lipid content extraction 2.1b: Agricultural
wastes
and transesterification

3.2-Pretreatment and Plants products with

Stored energy 2
fermentation
Processing steps for sugars
3
or monomers releasing for biofuel

world required a lot of quantity of sugarcane bagasse and Brazil was also reported with high-level research efforts
other plant biomasses such as from grasses, crop residues, involving the utilization of effective biochemical routes.
forest or wood industries for the production of second-gen- Earlier, diesel synthesis was achieved by the use of chemi-
eration ethanol or other chemicals. Biodiesel production in cal conversion routes, but the uses of microalgal species for

13

biofuel production, especially biodiesel, are currently more
focused on solving many energy challenges (Gargalo et al.
2016; Pfaffingera et al. 2019; Choi et al. 2019).
Recently, fermentation processes in Brazil have been
improved with the best achievement of conversion of theo-
retical stoichiometric rate with 82% in 12–16 h to the current
average conversion rate of 91% in 8–9 h (Basso et al. 2008;
Cho and Kim 2019). This bioprocess is exhibited by the uti-
lization of large tank size of 0.5 to 3 million liter capacities
with rigorous recycling of the yeast cells or strains (90%),
which has resulted in a high density of biomass inside the
fermenter tanks with 10% to 17 w/v wet basis in very short
fermentation periods. Fed-batch around 85% and continu-
ous 15% modes of fermentation were operated for ethanol
production using baker’s yeast as a starter with the minimum
cost and more availability (Basso et al. 2008; Walls et al.
2019; Tan et al. 2019).
Clostridium thermocellum and Trichoderma reesei are
utilized as effective microbial agents for the degradation of
lignocellulose. The S. cerevisiae is reported for very poor
tolerance of ethanol concentration, e.g., 25% v/v or less
quantity, and it is known as the common fermenting organ-
ism. Pichia stipitis is reported as an effective yeast strain,
which is found more relevant to biofuel production, and it
has developed its ability for the utilization of pentose sugar
such as xylose in natural manners toward biofuel fermenta-
tion process (Silva De Souza et al. 2016; Henriques et al.
2018).
n-Butanol is utilized in conventional engines offering
higher energy output or very close value of the energy out-
put of petroleum with an easy transport convenience by the
existing pipelines as shown in Fig. 2. Genetically engineered
strain E. coli was used for the production of isobutanol and
other alcohols via the utilization of the pathway of a non-
fermentative nature with more readily adapted conditions
for its production on a large scale. Engineered strain E. coli
was employed for the heterologous gene expression, which
is taken from Lactococcus lactis and Bacillus subtilis strains.
Thus, effective utilization of efficient microbials can help
for synthesizing biofuels from renewable resources via the
mitigation of global energy scarcity and environmental pol-
lution problems (Sa et al. 2011; Kumar and Kumar 2017).
Transportation fuels as biofuel have been utilized glob-
ally as an alternative option to fossil fuel and feedstock
supply of lignocelluloses, and this is indeed an efficient or
effective way for the stimulation for local biofuel indus-
tries and the overall economy (How et  al. 2019). The
mean–variance model for biofuel utility maximization was
applied for the estimation of supply equations for crop
feedstock in Washington, D.C., in the USA with the con-
sideration of price risk and examination of comparative
results. These results have helped to make some important
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Environmental Chemistry Letters
decision-making implications for the farmers in the Wash-
ington, D.C., area for biofuel production from the agri-
cultural masses or biomass feedstocks and different forms
of biofuels as shown in Table 2. In various oilseed crops,
canola cultivation is reported to be the highest oil yielding
crop with a limited quantity for many years in Washington
area, but some crops such as mustard and safflower are
reported to grow with lower oil yields than canola (Zheng
and Shumway 2008; Dumortier 2016).
Advances in engineering processes for microalgal spe-
cies with synthetic biology applications are reported for
the improvement in biofuel production. Also, extraordi-
nary diversity with higher biomass productivity with bio-
mass quantity up to 50,000 kg per acre per year and oil
content has been reported. Here, photosynthetic efficiency
of the microalgal species was used to be better for the
optimization of light- and carbon-capturing systems via
the utilization of genetic modification or modern synthetic
biology tools (Ansari et al. 2019; Yao and Shimizu 2013).
Moreover, the metabolic engineering technique utilization
microalgal species have uplifted the biosynthesis of lipids,
terpenoids or ­H 2 contents. Here, several challenges are
identification of strategies for crop protection strategies,
improvement in harvesting or oil extraction processes for
increased productivity of biomass and oil contents. These
challenges can be solved by applying the techniques of
genetic, molecular or synthetic biology techniques that are
well established for enabling the algal biofuel via making
economical-level competition with the fossil fuels (Ansari
et al. 2019; Yao and Shimizu 2013; Raheem et al. 2018;
Machado and Atsumi 2012).
The biosynthesis of biofuels in Sweden was found to
increase significantly over the past few years via reducing
fossil fuel dependency as well as the mitigation of green-
house gas emissions. Current methodological guidelines
were used for the assessment of greenhouse gas savings
from the use of biofuel. Other benefits such as electric-
ity and waste heat were found from biofuel biosynthesis
(Mahbub et al. 2019; Rassoulinejad-Mousavi et al. 2018).
Aggregation of the environmental performance of these
multifunctional systems can help in the assessment of
impacts and benefits from the relevant production pro-
cesses in Sweden via improvement in the decision-mak-
ing process for biofuel producers and policymakers in the
transition to a bio-based process. A road map for biofuel
production portfolio in Swedish with the development of
future bioenergy production scenarios via life cycle assess-
ment methodology was developed for the assessment of
the environmental performance of the production pro-
cesses (Mahbub et al. 2019; Rassoulinejad-Mousavi et al.
2018; Jong et al. 2017; Swain and Karimu 2020). Some
examples of biofuels are explored in this review.
Environmental Chemistry Letters
Algal biofuel and biodiesel development
On an industrial scale, microalgal biofuels production has
been reported using life cycle assessment (LCA) methods
with lesser energy inputs in biofuel development processes;
the least quantity of greenhouse gas emissions from the algal
biodiesel utility can be available in an automobile engine.
For microalgal biomass drying processes, the least energy is
needed in subsequent downstream processing steps (Jorquera
et al. 2010). Uses of an oil extraction process can be done
from the homogenization with heat recovery from the wet
biomass. In the cultivation phase for microalgal biomass in
raceway ponds, the most energy demand is fulfilled from the
electricity and it is utilized for circulating the algal culture
strain with energy fraction from 22 to 79%. The embodied
energy fractions with 8–70% in pond system construction
are required for algal biomass culturing (Kothari et al. 2017).
The embodied energy in the nitrogen fertilizer can be
used in a substantial contribution to the energy demand for
the cultivation phase energy faction of 6–40%. Normalized
PBR systems are reported to consume more energy than they
can produce. Dewatering and biomass drying processes are
less important than energy consumption in cultivation and
harvesting processes. PBR systems can provide greater
algal biomass concentrations via consuming more amount
of energy at the cultivation stage. Types of PBR systems are
reported to consume different amounts of energy, and these
are energy fraction for tubular PBRs (86–92%) and flat plat
PBRs with 22% (Jorquera et al. 2010; Kothari et al. 2017).
Microalgae biofuel production was reported for the
utilization of aquatic and terrestrial systems, and these
biofuels have shown the emerging strategy at the global
level for reducing the dependency of fossil fuel energy and
its potentials for the promotion of green environment to
Fig. 3  Algal biofuel synthesis.
The first step explains the algal
species nature, the second
shows the capacities in salty or
freshwater, the third explains a
laboratory or open pond condi-
tions, the fourth explains stored
lipid content, the fifth explains
thermochemical pretreatment
process for conversion of lipid
to biodiesel in form drop-in
hydrocarbon, and the final or
last part explains utilization of
electric power or heat energy
help to minimize the international environmental problems
such as global warming, climatic change or other earthly
challenges without affecting the global evolution. Micro-
algal species is shown as a bioenergetic resource, and gas
capacity has solved the uneven quantity of energy supply
for developing countries; its bioprocesses for biofuels are
shown in Fig. 3 (Mahmoud et al. 2016).
Contextual strength, weakness, opportunity and threats
(SWOT) models were utilized for the assessment of needs
and promises of the prevailing situation in Nigeria, and
this model can strategically analyze the risk and potential
benefits of biodiesel biosynthesis from the microalgal spe-
cies (Bull et al. 2016). Potentials of optimization of bio-
process for the development of biofuels such as biodiesel
or bioethanol from SWOT approach are reported from the
microalgae in Nigeria. SWOT analysis can provide pos-
sible benefits, derived from the development of algal bio-
fuels within the premises of possible setback without the
hindrance of scientific progress during the developmental
processes (Bull et al. 2016; Elegbede et al. 2017).
Microalgae biofuel has shown the least amount of C ­ O2
emissions such as low to zero quantity with the creation
of prospects of trading carbon emission as found different
from fossil fuels or non-renewable energy sources. The
produced biofuel and biodiesel from the algae species have
shown the high value of octane number (CN) with the
energy nature of carbon–neutral, self-reliance supply at the
commercial scale via a reduction in the global warming.
These biofuels have upgraded the life of the low-income
status of rural people with other benefits. The biofuel util-
ity and effectiveness were reported with the possessing of
good fuel quality than the non-renewable energy sources
(Elegbede et al. 2017; Das 2017).
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Nigeria has developed a broad range of prospective for the
biosynthesis of micro- or macro-level algal biofuels such as
biodiesel or bioethanol, which has shown the vis–vis water,
sunlight, space and different species of algae. Macro- and
microalgae biofuel production has shown more amount of
carbon emission consumption during their growth, and these
can be shown via more amount of carbon capture ability and
sequestration process of it is found to be a good strategy
for the reduction in global warming (Adegoke et al. 2018).
These algae species can be grown in any medium location
and different environmental conditions such as wastewater
sewage plants, freshwater, marine or brackish water without
competing to food land requirements. Invariable land qual-
ity and space are not needed for algal biomasses cultivation
with a generation of many value-added products for different
applications in industries and medical fields with a reduction
in biofuel cost (Adegoke et al. 2018; Gendy and El-Temtamy
2013).
Microalgae species have many challenges such as least or
minimum lipid yield under limiting growth conditions and
slower growth rate of high lipid contents of algal species.
These can be solved by the biotechnological interventions
to create major advances in algal species or strain improve-
ment strategy for the production of biofuel at commercial
industrial scales. Microalgae culturing has been reported by
carrying out wastewater treatment and ­CO2 sequestration
processes without the need for agricultural land or ecologi-
cal landscapes (Sharma et al. 2018a, b).
Efficient transformation toolbox has been applied to
increase the lipid accumulation via regulation of key enzyme
biosynthesis during the lipid synthesis. These have helped
via blocking of the competitive metabolic pathways with
pyramiding genes that have enabled in high cell biomass
cultivation under nutrient-deprived conditions or other envi-
ronmental stresses (Mathimania and Pugazhendhi 2019).
Furthermore, controlled by an upstream regulator of targets,
the transcription factors and microRNAs have helped in lipid
quantity enhancement in algal species (Mathimania and Pug-
azhendhi 2019; Sharma et al. 2018a, b).
Microalgal species such as Chlamydomonas are shown
to accumulate starch bio-component as the primary form of
energy storage via redirection of triacylglycerol (TAG) due
to the adaption in carbon shifting process by maintaining
membrane integrity. TAG molecules are found to backup the
catabolism process by releasing fatty acids (FAs) with stress
reversal and also for membrane synthesis (Yang et al. 2018).
This molecule is acting as a sink for channeling the excess
energy and reductive equivalents’ power. These microalgal
species have shown the presence of unique lipids such as
betaines with a good advantage in adaptation in nutrient-
limiting conditions as higher plants shown to the utiliza-
tion of phosphate-associated lipids with a central role in the
maintenance of membrane integrity (Work et al. 2010).
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Environmental Chemistry Letters
Microalgae have contained the distinct nature of acyl
groups in its storage lipids. There are reports on two orga-
nelle pathways for lipid storage, and it has shown the pres-
ence of 16-carbon chain acyl groups in the SN-2 position
with glycerol backbone in lipid, derived from plastic path-
way, and presence of 18-carbon chain acyl groups in the
same position in ER-generated lipids (Nedbalová et al. 2016;
Cherubini et al. 2011).
High capacity of C ­ O2 fixation in microalgal species is
shown to accumulate different quantities of bio-components
(such as lipids, proteins and carbohydrates) depending on
their growth conditions, and their biomasses represent very
good and enough source of fermentable sugars for the third
generation of bioethanol biosynthesis. The microalgal origin
carbohydrates utilization was also reported for bio-ethanol
biosynthesis and its production by following three main
steps, e.g., pretreatment, saccharification and fermentation
(Hwang et al. 2016; Kumar et al. 2019a, b; Anto et al. 2019).
The pretreatment process has helped to increase the acces-
sibility of intracellular sugars, which played an important
role in improvement in the overall efficiency of bioethanol
production. Chemical, thermal, mechanical, biological or
their combinations have been applied for promoting the cell
disruption, extraction facilitation and modification in car-
bohydrate structures as well as fermentable sugars produc-
tion (Hwang et al. 2016; Kumar et al. 2019a, b; Cheng et al.
2013).
Quantification of starch and total other sugars in microal-
gal biomass can be done for bioethanol production. Micro-
algal species have shown their autotrophic or heterotrophic
nature with the capability of synthesizing large amounts of
biomass containing lipid, protein or carbohydrates, depend-
ing on the species or its growing conditions as shown in
Fig. 4 (Oliveira et al. 2017; Al-Fadhli et al. 2019; Alam
et al. 2015). Microalgae species are made up of the main
components such as protein, carbohydrates, lipids, ashes or
acids among other compounds without lignin. Their cell wall
mainly consists of cellulose component containing pectin
and sulfated polysaccharides. Its plastids are found to con-
tain intracellular starch in the range of 20–50%. Majority
of lipid components (20–60%) are present inside the cells,
whereas protein concentrations are found between 20 and
50% depending on the biochemical compounds during the
growth conditions and culture medium (Ho et al. 2013; John
et al. 2011; Zollmann et al. 2018; Chemodanov et al. 2017).
Bioethanol and biohydrogen have been reported as biogas
and biomass-based energy, and their syntheses can enhance
the cost-effectiveness process by providing an opportunity
for sustainable fuels in rural community energy needs. A
substantial amount of TAGs such as storage lipid compo-
nents under photooxidative stress or other adverse environ-
mental conditions have been reported in microalgae species
cultivation in photo-bioreactors or open pond system with
Environmental Chemistry Letters
Calvin Cycle for Acety-Co A
glucose synthesis
Chloroplast
Endoplasic reticulum
Algal cells
Biodiesel Lipid droplet Triacylglycerol
Fig. 4  Glucose contents utilization for lipid content synthesis during
algal species cultivation. This figure shows the different biochemical
reactions in chloroplast cell organelle that showed glucose synthesis
from solar energy and then glucose is degraded into acetyl coA, and
then it is utilized in fatty acid, acetyl-acyl carrier protein (ACP) and
high biomass or lipid yield and multiple applications (Mano-
lis et al. 2019; Méndez et al. 2019). Microalgal species can
provide renewable biofuels as well as different types of bio-
products of high values as shown in Fig. 4. It offers for high
efficiency of a photosynthetic process or biomass production
with a faster growth compared to any other energy crops. For
future needs, biofuels can, therefore, be ideally fueled with
the creation of high benefits at the environmental, economic
or social levels worldwide; these can reflect the energy effi-
ciency with the plan of road map for industries via the third-
generation biofuel production (Manolis et al. 2019; Méndez
et al. 2019; Hemaiswarya et al. 2012).
Biohydrogen development
Biohydrogen ­(H2) fuel is produced in biological processes
and is reported as clean fuel energy that can be readily bio-
synthesized from certain kinds of biomasses. The H ­ 2 bio-
production has many challenges such as storage and trans-
portation of non-condensable gases. Recently, biohydrogen
biosynthesis has received significant attention as a future- or
next-generation energy carrier with its sustainable nature
fuel as discussed in Table 1 (Buitrón et al. 2017). The H ­ 2
production has been reported by certain biological routes
such as photobiological, dark fermentation, thermochemi-
cal and enzymatic processes. Therefore, H ­ 2 generation can
be achieved via the utilization of biological wastes as a
potential source through the biological processes with much
considerable interest via opening up a new avenue for the
utilization of renewable and inexhaustible energy sources
(Mirza et al. 2019; Lu et al. 2019).
Fatty acids Acetyl- ACP
synthesis
Free-Acetyl-
ACP
Dihydroxyacetone
phosphate
Glyceraldehyde 3-
phosphate
free acetyl synthesis in chloroplast. Later, these intermediates moved
to endoplasmic reticulum cell organelle, and then, it is converted to
triacylglycerol (TAG) or lipid droplet. Later, it is converted into bio-
diesel as final products
Biohydrogen ­(H2) production has been reported through
wastewater remediation via the emphasis of fermentative
routes such as light energy-driven and non-light energy-
dependent processes, and this was achieved via enclosing
wastewater system and mixed culture application as the
focal points. These bioprocesses are reported to consider
important process parameters on process efficiency (Cho
and Hoffmann 2017). However, the never-ending limita-
tions come across during these processes, due to lack of
process operation description and economically viable pro-
cesses. Stress is also reported as an inappropriate descrip-
tion of biocatalyst pretreatment methods in mixed cultures
with wastewater usage. The H ­ 2 fuel can thus be deemed for
high designed energy carriers for the future having a high
energy yield (Cho and Hoffmann 2017; Preethi et al. 2019;
Dumortier 2016).
A clean energy carrier property of biohydrogen ­(H2) was
reported with high energy yields and combustion of ­H2 fuel,
but the major by-product is only ­H2O molecule emission in
our environment. ­H2 fuel was reported with attractive and
renewable characteristics as its bio-production can be devel-
oped from a variety of biological routes. H­ 2 generation has
been reported by various modes via biological routes, and
these are categorized into various groups, including photo-
biological fermentation, anaerobic fermentation, enzymatic
or microbial electrolysis. Hydrogen production can also
be possible by a combination of the above processes (Kim
et al. 2018). Evaluating the biological routes for efficient
and improved hydrogen production has been reported by the
application of bioaugmentation, multiple process integration
and also microbial electrolysis processes; these processes
can improve the bioprocess efficiency via addressing the
13

Table 1  Biofuel biosynthesis from biowaste materials
Biofuels Substrates
Hydrogen production Water
Sustainable hydrogen production Grass Miscanthus sinensis
Bioethanol Miscanthus (sacchariflorus or
giganteus or lutarioriparius) as
cellulosic biomass
Bioethanol Dedicated energy crops and resi-
dues (Giant reed, elephant grass
and Miscanthus sacchariflorus)
and soybean litter and rice husk
Biohydrogen Acetate with increasing the phos-
phate buffer concentration
Hydrogen Waste biomass
Low-cost hydrogen production Water electrolysis
Hydrogen production Solar water splitting
industrial-level applications (Larsson et al. 2016; Hu et al.
2018).
Molecular hydrogen (­ H2) was reported as a clean nature
of burning fuel with a high-energy-rich high-energy com-
pound and was used mainly in industrial processes as a
reducing gas. Earlier, hydrogen was reported to synthesize
basically by the steam reforming process of natural gas,
which is non-renewable fuel, but biosynthetic strategies were
found to associate with poor yield and high cost (Hibino
et al. 2018). The application of an electrochemical driving
force was found in microbial electrolysis cells, which was
reported to improve the hydrogen yield in biological reac-
tions. MEC performance was influenced by the nature of the
materials used in electrodes, designing of reactors, microbial
strain consortia and substrates as experimental parameters.
Bio-electro-synthesis as promising technology was used at
industrial or commercial scale for the production of hydro-
gen ­(H2). Here, slow kinetics as well as consumption of
hydrogen in the biofilm was also reported as the bottlenecks
and complex feedstocks with the mixed nature of biofilms
(Vidales et al. 2019; Montpart et al. 2014; Paz-Mireles et al.
2019).
Bioenergy and resource recovery was achieved via the
utilization of microbial electrolysis cells (MECs), and MECs
are reported as emerging or advanced technology for waste
treatment processes. MECs have the capability for convert-
ing any biodegradable nature of waste into biohydrogen
­(H2), biofuels or other value-added bioproducts. However,
the efficacy of MECs can be varied with significant quantity
based on different types of substrate utilization or different
operational conditions (Li and Chen 2018). MECs design
13
Environmental Chemistry Letters
Methods/processes References
Alkaline water electrolysis, PEM Nguyen et al. (2019), Kovač et al.
water electrolysis and high-tem- (2019)
perature water electrolysis
Intermediate-temperature elec- Ito et al. (2018)
trolysis
Pretreatment and enzyme sacchari- Cerazy-Waliszewska et al. (2019)
fication with fermentation
Cellulose solvent-based lignocel- Ge et al. (2011)
lulose fractionation (CSLF)
pretreatment and enzymatic (cel-
lulase) hydrolysis. Self-flocculat-
ing yeast strain (SPSC01)
MEC–MFC coupled system Sun et al. (2008)
Direct electrolysis at intermediate Hibino et al. (2018), Cho and Kim
temperatures (2019)
Membraneless electrolyzers Esposito (2017)
Photovoltaic–electrolysis Jia et al. (2016)
application with their niches (bioprocess pathways or MEC
technology challenges) has been reported in integrative
waste biorefineries with a critical analysis of the perfor-
mance of MECs system with regard to hydrogen produc-
tion rate, yields and energy efficiency under the influence
of a variety of substrates, applied voltages and other critical
parameters. Mutual benefits have been reported between the
MECs and dark fermentation, which are reported as viable
approaches for the efficient nature of hydrogen production by
the utilization of renewable biomasses (Li and Chen 2018;
Quéméner et al. 2019).
Utilization of eukaryotic algae species was discovered
for the production of a renewable nature of fuels including
several bioenergy carriers such as alcohols from starches,
diesel from lipids and fuel cells from hydrogen. Compared
to terrestrial’s biofuel feedstock, microalgae species have
converted solar energy into biofuels with higher photosyn-
thetic efficiencies and these have the capabilities for thriving
in saltwater systems (Sharma and Arya 2017). Considerable
progress in microalgae species has been made by identify-
ing relevant or closed bioenergy genes and their pathways
through advanced or powerful genetic techniques applied to
engineering via the target-oriented disruption of endogenous
or same-origin genes or different-origin or transgene expres-
sion. These areas are rapidly advancing with their ability to
genetically optimize for the production of target biofuels,
including biohydrogen synthesis (Kumar et al. 2019a, b;
Shobana et al. 2017; Yun et al. 2019).
Microbial cells can also produce electric power current
inside the microbial fuel cells (MFCs) with an exhibition
of potential for practical applications as well as large-scale
Environmental Chemistry Letters
harvesting device development by utilizing microbial energy.
At a scale, standard polymer electrolyte membrane (PEM)-
based fuel cell technology has been utilized for the supply of
fuel without the need for purification with the maintenance
of the operation process at the ambient temperature by utiliz-
ing the biological materials (Mayur et al. 2018). Biological
catalyst agent has the capacity for self-regeneration power in
the synthesis of electric power during wastewater treatment
using MFCs. Anaerobic natures of treatment techniques
were used in wastewater treatment via the development of
energy harvesting processes. Varieties of designs for MFCs
are available in the literature with their rapidly expanding
applications over the past few years by utilizing biocompat-
ible materials and engineering designs (Mayur et al. 2018;
Sun et al. 2017).
Production of sustainable energy and clean water via
advanced wastewater treatment processes can be achieved
by the utilization of microalgal species in our advanced
society. Renewable nature of algal biomass feedstock for
biofuel synthesis can found via the treatment of wastewater.
Water composition was used as a growth medium (Zhan
et al. 2017; Kazemifard et al. 2019). Algal biomass has the
capability to resilience toxic emerging contaminants with
highly concentrated organic materials containing wastewater
such as organic nitrogen, phosphorus and saline compounds.
This has shown an excellent or high level of absorption of
emerged heavy metals or contaminants. Massive microalgae
culturing in wastewater has been the driving force for the
promotion of its utilization as a feedstock for biofuel utiliz-
ing algal biofuel technology on a commercial scale (Sun
et al. 2017; Zhan et al. 2017; Kazemifard et al. 2019).
The utilization of an integrated approach with microalgae
species was found in wastewater treatment via the consump-
­ O2 gases as well as biofuel production. The use of
tion of C
microalgae species with synergistically coupled with C ­ O2
consumption has been reported in advanced processes of
wastewater treatment with biofuel production in a sustain-
ability mode via using the life cycle analysis. Advanced
development in algal biomass cultivation and pretreatment
of biomass hydrolysis could be helpful in biofuels synthe-
sis. A low quantity of ­H2 production rate was reported in
the photosynthetic microbial cells due to the presence of
oxygen-sensitive hydrogenase enzyme in chloroplast orga-
nelle, and oxygen-sensitive hydrogenase in the chloroplast
and photosystem (PS) II have been reported to release O ­ 2
(Woertz et al. 2014).
Advanced‑ or second‑generation biofuel
Biofuels production fulfills total energy fuel requirement
with a broad spectrum of fossil fuels from gasoline to kero-
sene, and advanced biofuel can exhibit a critical role in the
replacement of fossil fuels. Cellulose-based ethanol and
some other biofuels can be applied as an energy source for
heavy-duty vehicles, ships or aviation as a transporting sys-
tem (Buijs et al. 2013). The advanced- or second-generation
biofuel was applied as a drop-in fuel source in the existing
internal combustion of the engine in the transport system,
which can be produced from the yeast cell factory Saccharo-
myces cerevisiae. Higher carbon chain alcohol-based biofu-
els such as n-butanol, isobutanol, sesquiterpenes (farnesene,
bisabolene), fatty acids and ethyl esters (biodiesel) as
advanced biofuels have been reported to improve the bio-
synthesis in the metabolically engineered S. cerevisiae for
the production. Some examples of biofuels and their produc-
tion are given in Table 2 (Buijs et al. 2013; Singh and Verma
2019; Jrad et al. 2020).
The economic growth of any country is strongly depend-
ent on the availability and price of sustainable nature fuels.
Recently, different parts of the world are putting many
efforts for decreasing the consumption of fossil fuels since
biofuels could be the main solutions for achieving more
economic growth at the global level. Advanced bio-ethanol
producer yeast Saccharomyces cerevisiae has been reported
to be important among the biofuel producer species (Walls
et al. 2019; Tan et al. 2019; Mamadzhanov et al. 2019). It
has never existed before the type of challenges can now be
solved with yeast biotechnology for the generation of future
biofuels at large scales via the utilization of sustainable
feedstocks without interfering with the production of food.
Sustainable feedstocks are not generally the traditional or
conventional carbon sources for the organism of Saccharo-
myces cerevisiae. However, the current trend in developing
biofuels from the engineered S. cerevisiae was utilized as
drop-in fuels for the existing internal consumption engines.
Production of designed biofuels in cost-effective mode was
also reported by applying Saccharomyces cerevisiae-based
cell factories (Walls et al. 2019; Tan et al. 2019; Kuo et al.
2019).
Engineered Saccharomyces cerevisiae can produce
advanced biofuels via the utilization of simple sugars with
high yield, and precursors of the drop-in fuel are derived
from the energy-rich fatty acids. Engineered microbial strain
can synthesize the drop-in fuels indirectly to offer scalable
and having control capability with the economic route for
fatty acid chemicals for bioenergy synthesis. Long-chain
hydrocarbon ester has been reported as diesel fuel, and
engineered yeast strain or other microorganisms via synthe-
sis of their cell membrane can also synthesize similar types
of fatty acids. Engineered yeast strain also can produce a
high quantity of fatty acids with the elaboration of a regula-
tory or feedback system via preventing excessive accumu-
lation of these building blocks (Fernandez-Moya and Da
Silva 2017). Department of Energy (DOE) Joint Bio-Energy
Institute has promoted the native highly regulated nature for
machinery for fatty acids with new high-intensity promoters.
13


Table 2  Conventional and advanced biofuel production from plant products or their wastes
Biofuel nature Biofuel examples Concentration References

13
Conventional Ethanol from sugars and starch in simultaneous saccharifica- Raw cassava flour hydrolyzed by enzyme PoGA15A that Xu et al. (2016)
tion and fermentation produced ethanol (57.0 g/L) and yield (93.5%) after 36-h
fermentation by Pichia pastoris
Conventional Xylose (25 g/L) and glucose (35 g/L) used for ethanol pro- Metabolically engineered Escherichia coli strain MS04 pro- Parra-Ramírez et al. (2018)
duction duced the 30 g/L ethanol from glucose and xylose sugars
Biodiesel from pure vegetable and synthetic vegetable oil 80% yield of biodiesel from transesterification of vegeta- Santana et al. (2012), Bourgeois et al. (2015)
ble oil (triglycerides) using supercritical ethanol on ion
exchange resin catalyst
Conventional Biodiesel from waste vegetable oils via its pretreatment with Biodiesel production, e.g., yield 97%. Methanol-to-oil molar Elkady et al. (2015)
transesterification process (120 °C) ratio (12:1) with 1% NaOH and tetrahydrofuran/methanol
volume ratio (0.3%)
Conventional Calcined eggshell utilized in the transesterification of vegeta- Calcining eggshell, e.g., 9 with 3% catalyst; at 800 °C for 2 h Kara et al. (2019), Abukhadra et al. (2018)
ble oil with methanol to produce biodiesel is used for yield increased biodiesel synthesis
Conventional Bioaugmentation of granular anaerobic sludge digesting Increased methane yield by 11% is reported by convert- Doloman et al. (2019)
mixed algal biomass in batch-scale reactors ing organic substrate into biogas in anaerobic digestion.
Addition of an algalytic bacterial mixture to the granular
consortium has increased methane formation
Methane yield and production rate in repeated batch bioaug- Biogas production, e.g., 5% and 10% methane, is reported Yang et al. (2016), Xiao et al. (2019)
mentation pattern from enriched microbial consortia and bioaugmentation
techniques
Advanced biofuel Chemocatalytic process used cellulosic ethanol in continuous Tandem reactions of cellulose to ethylene glycol and then to Yang et al. (2019)
operation and scalability ethanol. This process reported yield of ethanol, e.g., 43.2%,
in the presence of multifunctional catalyst Mo/Pt/WOx
Advanced biofuel Biomass of algae contain 20–80% oil contents that produced Biodiesel areal productivity is found high (19–25 L of BD/ Baliga and Powers (2010), Aresta et al. (2005)
diesel m2/yr), and photobioreactor is used for algae production
Advanced biofuel Microalgae Phormidium autumnale used sucrose as an exog- Algal cell oil productivities of 18.9 mg/L in steady-state Siqueira et al. (2016), Ishak et al. (2019)
enous carbon source conditions are reported for biodiesel synthesis, e.g., ester
content of 99.8% and cetane number of 58.5%
Advanced biofuel Hydrotreated vegetable oil from waste materials Hydrotreated vegetable oil exhibited better viscosity, density, Zeman et al. (2019)
cold filter plugging point of fuel blend and flash point and
is used for fuel energy
Advanced biofuel Biomass-to-liquid (BTL) processes for the production of Energy efficiency and production costs of the BTL designs Dimitriou et al. (2018), Gonçalves et al. (2013)
liquid hydrocarbon fuels. are reported from 37.9 to 47.6% lower heating value (LHV)
with a cost reduction of €17.88–25.41 per GJ of produced
fuels
Advanced biofuel Bio-SNG production from an indirectly heated circulating Aromatic compounds converted into Bio-SNG with olefin Haro et al. (2016), Chauhan et al. (2009)
fluidized bed gasifier hydration and hydrodesulfurization combined at high tem-
perature in hydrodesulfurization unit. Later, methanation
section is modified with H ­ 2S and ­CO2 removal
Environmental Chemistry Letters
 Environmental Chemistry Letters

Engineered cellular machinery can develop a new route for

fatty acids from the manufacturing of cell membrane, and
free fatty acids can be transformed through the industrial

Im-orb and Arpornwichanop (2020), dos

process to drop-in biofuels. Engineered microbial strains can

Im-orb and Arpornwichanop (2020)

enhance the production of free fatty acids (500-fold more)
compared to the native or wild strain utilization (Runguphan
and Keasling 2013).
Second-generation ethanol plants are shown to be the

Santos et al. (2017)

Salleh et al. (2019)

major significant stages in the construction of effective S.

cerevisiae strain for the utilization of fermentation processes
Martín (2017)

and lignocellulosic hydrolysates from the agricultural resi-

References

due or energy crops. Novel industrial contexts of microbial

strains yeast can be developed via metabolic engineering
strategies that can address the challenges. Proof-of-concept
the transformation of lignocellulosic biomass into DME via
the gasification process. Later, syngas is used by reforming,
CO2 is captured to produce DME via hydrogenation and also
showed 11% in a total improvement and 72% in situ recov-

studies have been applied for the construction of robust

and production of methanol (rate ~ 0.23 kmol h−1) with an
synthesis (IBPGM) process used for rice straw hydrolysis

(0.09 kmol h−1) from rice straw with an energy efficiency

2.88 g/L of biobutanol is produced from separate hydroly-

scrubbing or carbon capture technologies to get the final

sis and fermentation (SSF) with a yield of 0.23 g/g that

microbial strain platforms to facilitate industrial applica-

Integrated biomass pyrolysis, gasification and methanol

tions (Lee et al. 2017). Currently engineered and industrially

important S. cerevisiae strain was shown to achieve the pro-
IBPGM process is applied for bio-oil production

jected outputs from the first large-scale second-generation

ethanol plant, and some examples of biofuels are discussed
in Table 2. Academic- as well as industrial-level yeast strains
can help to strengthen the economic value for the second-
generation ethanol synthesis by achieving improvement
energy efficiency of 60.7%

in the fermentation kinetics and product yield of cellular

ery using a gas stripping

robustness under the fermentative process conditions (Dijk

et al. 2019).
The environmentally friendly microbial fermentation pro-
form of DME
Concentration

cess has been widely utilized for fatty acid-derived advanced

of 60.7%

biofuels production from the renewable feedstock. A combi-

nation of Saccharomyces cerevisiae and Yarrowia lipolytica
has shown more robustness as well as high tolerance capac-
ity for fermentation inhibitors and phage contamination;
Advanced biofuel Use naturally captured ­CO2 to form lignocellulosic biomass
Advanced biofuel Biobutanol production from lignocellulosic biomass of oil

this yeast strain has attracted tremendous attention in recent

years (Hu et al. 2019). For the biosynthesis of fatty acids
derived biofuels, few representative examples are fatty acids,
Advanced biofuel Methanol production from the IBPGM process

fatty acid ethyl ester, fatty alcohol or alkanes. Metabolically

engineered yeast strain has shown much improvement in
Advanced biofuel Bio-oil production from IBPGM process

fatty acid-derived biofuel via identifying the bottlenecks

that converts dimethyl ether (DME)
palm empty fruit bunch (OPEFB)

genes (Hu et al. 2019; Sheng and Feng 2015).

Development of an S. cerevisiae XUSE strain with high
performance in xylose-fermenting capacity was reported
for high ethanol yield without inhibition of glucose sub-
strate. S. cerevisiae XUSE strain has been characterized at
Biofuel examples

the genomic and transcriptomic level, and this strain char-

acteristic could reveal about the isomerase-based engineer-
ing strategies for improved xylose fermentation (Hoang
et al. 2018). High xylose fermentation performance can be
achieved via further engineering and XUSE. Xylose-utiliz-
Table 2  (continued)

ing engineered S. cerevisiae strain can be a promising plat-

form the strain for lignocellulosic biorefinery via conversion
Biofuel nature

of xylose substrate into ethanol at an efficient level and high

ethanol yield of 0.43 g/g. Evolutionary and combinatorial
clustered regularly interspaced short palindromic repeats

13

(CRISPR)-cas9-mediated engineering strategy could help in
obtaining newly refactored isomerase-based xylose substrate
fermenting strain yeast XUSE. Genomic and transcriptomic
analysis in XUSE strain can provide information about the
important mutation with changes in gene expression and can
be responsible for the enhanced rate of xylose fermentation
recognized in XUSE strain (Hoang et al. 2018; Matsushika
and Sawayama 2011; Romaní et al. 2015; Tani et al. 2017).
Some biofuels’ utility discussed in this review is shown in
Table 1.
Biofuel utility
Biofuel utility in the transportation sector
Various types of energy sources including biofuels are used
in the transport sector in the USA, which has shown 28% of
biofuel consumption of the total energy in 2018. The con-
tributions in the form of biofuels (ethanol and biodiesel) are
reported in the transportation of people and goods from one
place to another. Ethanol and biodiesel are the first biofuels
reported to be used in automobiles that can replace later
the gasoline or diesel of petroleum products. Presently, bio-
ethanol and biodiesel are blended in different ratios with
gasoline or diesel fuels for the motor engine (Zhang and
Chen 2015). In the year of 2018, total biofuels consump-
tion was reported to be up to 5% of the total energy in the
US transportation sector and ethanol biofuel was reported
to contribute 4%, while biodiesel contribution was 1% as
shown in Fig. 5, which shows the biofuel utility for different
applications. In the same year (2018), a report on differ-
ent energy sources or fuels was shown on the transportation
sector in the USA with gasoline petroleum (54%), distillate
gasoline (23%), jet petroleum (12%), biofuel (5%) and natu-
ral gases (3%) (Zhang and Chen 2015; Lenhart et al. 2016;
Shittu et al. 2019).
Different benefits at the level of energy security, socioec-
onomic or environment have been reported, and these were
the main reasons for the Indian government for supporting
biofuel production and its utilization. As regards the total
energy demand, India ranks the sixth position worldwide and
there was 3.5% of the world commercial demand in India
during 2001. Crude oil at the domestic-level production
reached a profound stagnate level with the rising of rapid
rate demand and crude oil with increased imports tendency
(Fulvio et al. 2019; Sotirov and Storch 2018; Sayed et al.
2012). During the period 2004–2005, India has done the
import of crude oils (95.86 million tonnes) amounting to
US$ 26 billion with a more expected import of crude oils
(98.26 million tonnes) with the values of US$ 41 billion in
2005–2006. Production of biofuels at an indigenous level
was shown as one of the options for the partial substitution
13
Environmental Chemistry Letters
of petroleum fuel with a reduction in dependency of
imported oils from other countries. Government of India
has planned for raising the plantation of Jatropha species on
a large wasteland area (0.4 million hectares), and National
Mission on Biodiesel (NMB) has been constituted with the
plan for biofuel feedstocks source as a part of ministry of
rural development under the nodal ministry (Fulvio et al.
2019; Sotirov and Storch 2018; Kumar et al. 2006; Röder
et al. 2019).
Biofuels utility in the transport sector has been an impor-
tant contribution with the development of some critical strat-
egies for improving the fuel security as well as the mitiga-
tion of climatic changes with full support to energy needs
to rural community development. By 2010, more quantity
of conventional biofuels (84 million tonnes) were gener-
ated based on food crop cultivation products such as starch,
sugars or vegetable oils, and in the future, 104 billion lit-
ers of biofuels can address the same percentage of global
demand (2.7%) for transporting engines as fuels (Raud et al.
2019; Harnesk 2019; Ramirez and Rainey 2019). For con-
ventional biofuels, biosynthesis (e.g., ethanol) as well as
biodiesel protein feed can deliver to the world. In the year
2010, the necessary amount of protein production was found
to associate with the conventional type of biofuel synthesis
and these were based on corn, cereals, canola or soybeans
utilization. These efforts were able to deliver more quantity
of protein feed (79 million tonnes) with the production of
protein-rich foods in more cultivation lands. Nearly 29 mil-
lion hectares of soybean crops were grown as a form of soy-
bean cake to fulfill the quarter of global demand (Mondou
et al. 2018; Kaza 2020). In addition, the biofuel production
chain of conventional mode has helped to fulfill the global
fuel demand as well as supplement more quantity of pro-
tein. The advanced nature of biofuel synthesis was found to
depend on the utilization of cellulosic feedstocks, various
waste organic compounds streams and algal species as large
potentials in the future (Mondou et al. 2018; Li, et al. 2018;
Mulholland et al. 2017).
The transport biofuel market has a share with less than
40% contribution. A higher proportion of the transport bio-
fuel market was shared in the interim periods of biofuel syn-
thesis (15–30 years) compared to the long-period biofuel
synthesis (e.g., above 30 years). In long-term fuel models,
models on the horizon and transport biofuels were substi-
tuted by the electric or hydrogen fuel model cars. Trans-
port sector biofuel utilization in a global economy model
has helped the future role in energy demands (Unrean et al.
2018; Schemme et al. 2019). Comprehensive global eco-
nomic modeling studies provided a portrayal of transport
biodiesel development in terms of their potentials and com-
petitiveness. More quantity as well as longer use of non-
fossil energy sources in the transport sector may help to
increase sustainable energy–transport interaction with the
Environmental Chemistry Letters

Bio-wastes materials Biofuels

from Municipal sites Bio-waste handling Preprocessing Anaerobic digester
via cans or vessel equipment

Biofuels
utility
Thermal energy or heat Gas grid
generation
Fuel generation

Compressed
Electric power generation natural Gas

1: Biowastes matters
2: Waste processing unit

A
Biofuels
utility
5:
3: Anaerobic
Biofuels 4: Biofuels digester
B operating
equipments
A: Gas grid
B: Heat generation
C D E C: Compressed natural Gas
D: Electric power generation
E: Fuel generation

Fig. 5  Biowastes materials utility for biofuels synthesis. This figure biogas or fuel production units such as anaerobic digester. The fourth
shows the biowaste matters in the municipal or commercial place site shows produced biofuels, and the final part shows the biofuel utility
in the first part, and it has gone to the waste processing unit to sepa- in different equipment for their functions
rate the organic matters in the second part. The third part shows the

system effects over the sector boundaries. Many reports are power generation, oil refinery and synthetic fuel synthesis
available on the development of several energy conversion (García-Olivares et al. 2018). There are various models,
technologies for carbon capturing and its storage (CCS) in which include several technologies for the synthesis of

13

alternative transport fuels and biohydrogen (­ H2), metha-
nol, dimethyl ether (DME), Fisher–Tropsch-based diesel
as bioethanol or biodiesel (García-Olivares et  al. 2018;
Anandarajah et al. 2013; Jin and Sutherland 2018).
Biofuel utility in the electric power generation
sector
Bioenergy electric generation and biofuel sub-technologies
have employed a total of 112,642 workers. Of them, only
7980 workers have been exclusively involved with bioen-
ergy or biomass electric generation technologies. A number
of women workers involved in bioenergy/biomass electric
generation technologies are somewhat lesser. Electric power
transmission and distribution normally occur with the grid
and the bulk of electricity transfer from power plant supply
to the center of the need point (Pick 2019). Transmission of
electricity and distribution with storage of electric power
can be done via making the entire network of power lines
and transmission of electricity to achieve from power gen-
eration stations to consumers. It can support other activities
from the power support and pipeline construction with fuel
distribution and transportation via electrical transmission
equipment manufacturer (Ram et al. 2019; Masmoudi et al.
2019). More amount of electricity generation is occurring
from steam turbines via using fossil fuels, nuclear energy,
biomass hydrolyzes, geothermal or solar-mediated thermal
energy. Other major technologies for electric power genera-
tion are reported from the utilization of gas turbines, hydro-
turbine wind turbine and solar photovoltaic cells (Shafiei
et al. 2017). Biomass, as a solid biofuel, contributed about
2% of the total US electricity during the year 2018, and
this was produced directly in the steam electric plants. Bio-
mass can be utilized via conversion into a gas, which can
be combusted in a steam generator, gas turbine or internal
combustion engine generator (Lenhart et al. 2016; Shittu
et al. 2019). South Korean utilities use biofuel oil for power
generation, and their governments have put more effort to
reduce the emissions of greenhouse gases by encouraging
the usage of cleaner energy. The energy ministry of South
Korea has revised petroleum business law from 2018, which
has permitted the power utility from the use of biofuels oils
as an alternative fuel source to generate electricity. Biofuel
oil power can make up only a small portion or fraction for
the needs of electricity in South Korea with more electricity
needs from the utilization of coal or nuclear power (Tylor
et al. 2016; Brigljević et al. 2019; Arifin et al. 2014). Wood
waste has been produced by the nearby fuels synthesizing
companies in California of USA, and 50-MW biomass power
plant is reported in the USA. These companies have utilized
the biopower or biomass power to generate electricity and
applied the system for direct firing, co-firing, gasification,
anaerobic digestion and pyrolysis processes (Liu et al. 2016;
13
Environmental Chemistry Letters
Goerndt et al. 2019). These small module techniques have
helped in the production of biopower. The company made
the arrangements for burning bioenergy feedstocks directly
and converted into steam energy that was normally captured
by a turbine system using the generator for the conversion
of steam energy into electricity (Verdezoto et al. 2019). The
utility of wood waste was reported for the production of
electric power and steam energy in paper mills. Coal-fired
power plants have been reported for the utilization of co-
firing system that helped to reduce greenhouse gas emissions
with gases such as ­SO2 and ­SO3 (Aghbashlo et al. 2019). Co-
firing systems have been utilized in high-efficiency boilers
system, and bioenergy feedstocks were used as supplemental
energy sources (Liu et al. 2016; Goerndt et al. 2019; Agh-
bashlo et al. 2019).
Biofuel utility in heat energy
In majority of cases, solid form of biofuel has been used
in heating energy production and wood as part of higher
plants is found in our planet in huge quantity and is used in
a both esthetic and practical method for heating purposes in
domestic needs or industrial energy demand like sugarcane
industry. The use of wood is to burn in home stoves as sup-
plement resources with other heating systems such as from
natural gas or electric power utilization (Shafiei et al. 2017;
Väisänen et al. 2016). Solid biofuels have raised the prices of
energy in recent years. Upward movement in the innovation
of industries and, with a research focus, liquid form of bio-
fuel were utilized with improved efficiency, reduced emis-
sions of greenhouse gases and enhanced convenience. About
91% of efficiency was reached in wood gasification boilers.
Wood pellets and wood have been reported with the heat
energy efficiency of 70% and 70–91%, respectively, with
their costs of US$ 2.16 and 1.20 per 1000 BTUs (Shafiei
et al. 2017; Chollacoop et al. 2013; Liu et al. 2019; Breetza
and Salon 2018).
The generation of biopower was achieved from dry bio-
mass combustion or gasification processes. In addition,
biogas (methane gas) can be biosynthesized or captured via
controlled anaerobic digestion. Agricultural biomass utiliz-
ing co-firing system had a low-cost system for the reduction
in greenhouse gases via improvement in cost-effective pro-
cesses with a reduction in air pollutants concentration from
the existing thermal power plants (Yue et al. 2014; Wood-
bury et al. 2018). Thermal energy system (cooling or heat-
ing) can be generated at a smaller scale for the utilization
in individual buildings, and these can be achieved through
direct combustion of wood pellets, wood chips and other
sources of dry biomasses (Solé et al. 2018). Operations for
combined heat and power (CHP) applications have shown
the efficient ways of utilization of biomass (80% potential
energy). During the biopower production process, waste heat
Environmental Chemistry Letters

Table 3  Biofuels utility for energy requirements in different sectors with a reduction in greenhouse gases/toxic chemicals worldwide
Biofuel utility Reduction in greenhouse gases/toxic Country References
chemicals

Cellulosic ethanol CO2 (2.11–2.30 kg or 90.9%); particu- Australia Jo et al. (2019).

(1 L) utilization late matter (size of less than 1 μm
(90%)
Benzene, 1,3-butadiene and PAH Australia Jo et al. (2019)
(polycyclic aromatic hydrocarbons)
up to 30–70%
Sugarcane butanol 55–18 ­gCO2–eq/M.J. sugarcane Brazil Pereira et al. (2015)
butanol
Corn butanol 79–122 ­gCO2–eq/M.J. corn butanol Brazil Pereira et al. (2015)
1.6 million gallons of biofuel (sustaina- Reduced opacity (visibility) and emis- Malaysia Karl (2011)
bly produced crude palm oil blended sions of nitrogen oxides ­(NOX) and
with palm stearin) sulfur dioxide (­ SO2) with 90 MW
oil-fired turbine electric power
capacity
Use of advanced biofuels (27% of total 2.1 gigatonnes (Gt) of ­CO2 emissions France Gonzalez-Salazar et al. (2016)
transport fuel) per year
Biofuels production (60–90 Mtoe) in 20% of all anthropogenic greenhouse Brazil and USA Fontes (2010)
2015–2030, respectively gases emissions with 80% of global
biofuels
Ethanol from sugarcane Greenhouse gas saving, Worldwide Fontes (2010)
Ethanol from poplar 87–96%
51–100%
Algae-derived biofuel CO2 emissions (50–70%) Worldwide Kommers (2013)
Biofuels like algal biodiesel and other Reduced greenhouse gas emissions Worldwide Hundt and Reddy (2011)
products with algae utilization of by 5%
66% of exhaust ­CO2
Corn-based butanol utility Reduction in greenhouse gases emis- Worldwide Yu et al. (2020)
sion by 32–48% and fossil energy
savings (39–56%)
Biomass-based 2-ethylhexanol (2-EH) Reduced greenhouse gases emissions Gomes et al. (2018), Huang et al.
by more than 85% compared to fossil (2018)
diesel
Ordinary gasoline blended with 0–150% reduction in greenhouse gases Brazil Macedo et al. (2008), Mahbub et al.
18–25% (v/v) of anhydrous ethanol, (2019), Rassoulinejad-Mousavi
known as gasohol et al. (2018)
Ordinary gasoline blended with legal 0–150% reduction in greenhouse gases Japan Mahbub et al. (2019), Rassoulinejad-
limit of ethanol blends is 3% (v/v) Mousavi et al. (2018), Cherubini
and Strømman (2011)

or steam energy can be used for providing the heat or chill
to nearby buildings or pipes (Yue et al. 2014; Woodbury
et al. 2018; Beagle and Belmont 2019; Bertrand et al. 2014).
Biofuel production has been reported from the agricul-
tural biomass wastes, and the most common types of bio-
fuels such as biodiesel, bioethanol and biogas have been
biosynthesized from the biomass utility. Plate heat exchang-
ers can be applied for heating or cooling the liquids or gases
via re-boiling or condensing processes in distillation col-
umns. During the recovery of renewable feedstocks (starch,
cellulose or sugars), wastes via heat exchangers have been
used for heating, evaporation or heat energy recovery pro-
cesses (Wang et al. 2019). Renewable feedstocks have been
reported with different components consistency and solid
contents fractions that are found to utilize in a different type
of heat exchanger for the biofuel production. Biodiesel can
be made from different types of vegetable oils, and the best
examples are soybean, sunflower or palm oil. These oils with
animal fats were used via chemical reaction applications
with methanol reagent for biodiesel production (Yrjol and
Paavilainen 2004). Heat exchanger applications are used for
various processes, and preheating, glycine heater or cooler
as well as methanol distillation has been the main processes
(Yrjol and Paavilainen 2004).
Biofuels are normally found in liquid or gaseous fuel
forms, and these can be biosynthesized via the utilization of
organic matter sources such as biomass crops, agricultural
waste or residues, and oils crops. Various sources of oils

13

crops products are palm oil, canola oils, soybean oils and
waste cooking oils, and these are outlined in Tables 1 and 3
(Poudel et al. 2017; Shafiei et al. 2015). In biofuels produc-
tion, various types of organic matters are used, of which
grease, food waste, animal residues or wastes, sewage and
landfill are the best examples. For instance, each tonne of
wood has been used in Swedish buildings, which can avoid
nearly three tonnes of ­CO2 emissions. The market for wood-
based heat and power was able to displace fossil-based fuel
oil that is largely saturated in Sweden with further wood
resources with the other biofuel production, including jet
fuel (Poudel et al. 2017; Moriarty et al. 2019).
Conclusion
For current and future scenario, sustainable biofuel utility
can be fulfilled for the worldwide energy needs in cost-
effective manners. Many nations of the world need more
awareness of biofuel utility in the transport sector as well as
electric power generation with its critical role in the miti-
gation of greenhouse gas emission as well as clean energy
development. Biofuel utility in electric power generation and
car fuel power in some countries has been reported with
more reduction in greenhouse gas levels up to 50%. How-
ever, biofuel developments can be achieved from food grain,
non-food grain or stored oil content in algal species with the
creation of more employment opportunities for the location
of people in the farming sector. Promotion in biofuel quan-
tity can be done via providing more subsidies on its produc-
tion cost by making its convenience and help in adoption to
local people for their energy needs.
Algal and non-food grain origin biofuel developments
can enhance the possibility of their utility in various sec-
tors for energy demands. Hydrogen as a clean fuel carrier
is found as a good option for the mitigation of greenhouse
gases and climatic changes. Microbial electrolysis cell
(MEF) is a rich source of electrochemically derived energy,
which can improve the ­H2 yield or its production quantity
in biological reactions. Sustainable advanced biofuel pro-
duction with wastewater treatment can be found in algal
species via producing clean water. At commercial scales, a
massive quantity of microalgae is utilized in the algal bio-
fuel development process. Production of high carbon chain
alcoholic biofuels such as 1-butanol or isobutanol, sesquit-
erpenes (e.g., farnesene or bisabolene) and fatty acid ethyl
esters (e.g., biodiesel) as advanced biofuels can be improved
via the utilization of metabolic engineering techniques in
S. cerevisiae or other microbial fuel cells. Biofuel utility
has been reported mainly in the transport sector, electric
power generation and heat energy production, and it has
mitigated the greenhouse gas emission of toxic gases to a
13
Environmental Chemistry Letters
great extent with the maintenance of natural resources to
maintain sustainability.
Compliance with ethical standards 
Conflict of interest  The authors declare that there are no conflicts of
interest.
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