Renewable and Sustainable Energy Reviews 58 (2016) 180–197

Contents lists available at ScienceDirect

Renewable and Sustainable Energy Reviews

journal homepage: www.elsevier.com/locate/rser

Microalgae biofuels as an alternative to fossil fuel for power generation

Jassinnee Milano a, Hwai Chyuan Ong a,n, H.H. Masjuki a, W.T. Chong a, Man Kee Lam b,
Ping Kwan Loh a, Viknes Vellayan a
a
Department of Mechanical Engineering, Faculty of Engineering, University of Malaya, 50603 Kuala Lumpur, Malaysia
b
Chemical Engineering Department, Universiti Teknologi PETRONAS, 32610 Bandar Seri Iskandar, Perak, Malaysia

art ic l e i nf o a b s t r a c t

Article history: Biofuels productions from microalgae received wide attention recently and have high potential to replace
Received 20 July 2014 fossil fuels. This paper served as a platform to critically review current production technologies of
Received in revised form microalgae, ranging from cultivation, harvesting, extraction and several biofuels conversion processes. In
14 October 2015
addition, due to the high photosynthetic efficiency of microalgae, mass cultivation of microalgae is
Accepted 17 December 2015
Available online 12 January 2016
believed to be able to efficiently reduce the carbon dioxide emission to atmosphere and thus, reducing
the impact of global warming. This is because microalgae have high growth rate and is able to develop
Keywords: maximum of 70% of lipid content within their cells depending on species. Apart from that, microalgae
Microalgae have the ability to survive under harsh condition and occupied smaller cultivation land area than other
Biofuels
land crops. The harvested microalgae biomass can be used for electrical generation, while its crude lipid
Carbon dioxide mitigation
can be used as transportation fuel as it has 80% average energy content of petroleum. In the present
Renewable energy
Alternative fuel paper, a detailed discussion to produce biodiesel, fuel gas, bio-oil, methane, hydrogen and alcohol from
microalgae biomass are also included. Besides, updated research, challenges and the way forward of
microalgae biofuels are also presented. In future, biofuels production from microalgae can be economical
viable at some scale, which is then profitable in terms of economics and also environment.
& 2015 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
1.1. Greenhouse gasses (GHGs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
1.2. Unsustainable energy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182
1.3. Renewable energy sources . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182
2. Biofuels as an alternative to fossil fuels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182
3. Biofuels production from microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
3.1. Microalgae cultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
3.1.1. Open pond systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184
3.1.2. Photo-bioreactors (PBR). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
3.1.3. Hybrid photo-bioreactors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
3.2. Harvesting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
3.2.1. Flocculation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
3.2.2. Filtration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
3.2.3. Gravity and centrifugal sedimentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
3.3. Extraction and purification for microalgae biomass. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
3.4. Conversion process and biofuels production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
3.4.1. Chemical conversion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
3.4.2. Thermochemical conversion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
3.4.3. Biochemical conversion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189

n
Corresponding author. Tel.: þ 60 16590 3110.
E-mail addresses: onghc@um.edu.my, ong1983@yahoo.com (H.C. Ong).

http://dx.doi.org/10.1016/j.rser.2015.12.150
1364-0321/& 2015 Elsevier Ltd. All rights reserved.
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197 181

4. Energy generation from microalgae biofuels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189

4.1. Microalgae biofuels for internal combustion engine . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
4.2. Electricity production by biomass from microalgae. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
5. Co-processes in microalgae cultivation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
5.1. Potential mitigation of CO2 emission by microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
5.1.1. Metabolism and photosynthesis of microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
5.2. Water treatment potential of microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
5.3. Microalgae biofuels: challenges and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
5.4. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195

1. Introduction microalgae biomass until the conversion technologies of micro-

The world population has experienced continuous growth
since the last 50 years, which directly resulted in a large increase
in primary energy consumption. In 2010, world primary energy
consumption grew by 5.6%, which is the largest percentage growth
in almost 40 years [1,2]. Hence, the world is currently facing two
detrimental challenges, which is energy crisis and environmental
pollution [3].
Energy crisis happened in the past decades due to the sub-
stantial reduction of unsustainable sources like fossil fuels.
Extensive use of fossils fuel for power generation and as trans-
portation fuel has caused high carbon dioxide (CO2) emissions to
the atmosphere and there is an urgent need to reduce its emission
to avoid detrimental impact of global warming. High energy
demand in future coupled with the concern of environmental
hazards, national security have heightened attention towards
production of clean liquid fuel, termed as biofuels, as a suitable
alternative source of energy [3].
Biofuels can be produced via various renewable feedstocks such
as edible oil (e.g. palm oil), non-edible oil (e.g. jatropha) and lig-
nocellulosic biomass (e.g. wood). The production of biofuels does
not only reduce the dependence on fossil oil trade, but also
reduces the uncertainties caused by the fluctuations in fossil oil
prices. The increase in fossil oil prices has caused more burden to
consumer, businesses and investor as it generates high competi-
tion against other countries which utilize biofuels or alternate
energy. Therefore, these countries that used biofuels can prolong
and maintain low inflation rate. Since fossil fuel is based on global
pricing, therefore the country that used such fuel for transporta-
tion, production, and all oil related products prices will fluctuate.
Biofuel is a renewable energy source which provide an assurance
to the economy of the country while promising to achieve a green
environment.
Plants have the ability to convert solar energy into chemical
forms though photosynthesis, in which CO2 is captured from the
atmosphere and oxygen (O2) is produced during the process.
Approximately 77 Giga tons of CO2 is fixed via photosynthesis,
leading to production of 100 Giga tons of biomass annually (lig-
nocellulose materials) [3]. Microalgae has higher photosynthetic
efficiency than other crops (e.g. plants or trees), which utilizes
algae biomass to biofuels and its advantages. Furthermore, a
comprehensive discussion of the carbon fixation and wastewater
treatment by microalgae are also included in the present paper. It
is hoped that this review paper will pave a way forward in rea-
lising microalgae biofuels production for energy security purpose
and sustainability of environment.
1.1. Greenhouse gasses (GHGs)
It is well known that the burning of fossil fuel has led to
greenhouse gases emissions such as carbon dioxide (CO2), sulfur
dioxide (SO2), nitrogen oxide (NOx) and carbon monoxide (CO).
The emission of these greenhouse gases has increased year by year
and this has caused the serious global warming effect [5].
The global energy-related CO2 emissions are estimated to rise
double the amount by 2035, or at the rate of 1.6 percent a year. In
year 2010, the emission of CO2 was 110 billion metric tonnes and
the emissions are forecasted to reach over 140 billion metric
tonnes of CO2 in year 2035. The rate of global CO2 emission by end-
used sectors from 1990 to 2035 is shown in Fig. 1 [6]. High level of
CO2 in the atmosphere impedes the passage of thermal infrared
radiation emitted from the surface of the earth back into the space
that consequently caused temperature of the earth to increase.
The Kyoto Protocol of 1997 called for a 5.2% reduction in
worldwide GHGs emission based on year 1990 and to meet the
agreed target. Therefore, a selection of effective technologies,
including chemical and biological CO2 mitigation technologies, has
been a focus on research to reduce the GHG emissions [4]. In
Malaysia, the amount of greenhouse gases increases every year.
Fig. 2 showed the projected CO2 emissions in Malaysia from 2000
to 2020 [7]. CO2 emission is estimated to double in 13 years from
year 2000 reaching 200 million metric tonnes, with approximately
8.8% increment annually.
160.0
140.0
Carbon dioxide emission

approximately 183 Gtons of CO2 to produce 100 Gtons of micro-

(billion metric tonnes)

120.0
algae biomass [4]. Therefore, microalgae biomass cultivation will
100.0 Others
help in reducing concentration of CO2 in the atmosphere at a faster
80.0 Industrial
rate than other land based crops, especially when microalgae are
60.0 Power Generation
cultivated under optimized environment conditions.
This paper aims to demonstrate an overview of the potential of 40.0 Transportation

biofuel production from microalgae biomass and the importance 20.0

of microalgae biomass as an alternative resource to reduce 0.0
1990 2000 2010 2015 2025 2035
dependencies on fossil fuel. Besides, the present paper also dis-
Year
cusses the overall process chain of microalgae biofuels production,
including cultivation, harvesting, extractions and purification of Fig. 1. Global CO2 emission from various sectors from 1990 to 2035 [6].
182
300.0
250.0
Carbon dioxide emission
(million metric tonnes)
200.0
150.0
100.0
50.0
0.0
2000 2004 2008 2012
Year
Fig. 2. CO2 emissions from various sectors in Malaysia [7].
5.4%
9.0%
0.8%
3.8%
2.6%
78.4%
Fig. 3. World primary energy consumption in 2014 [8].
1.2. Unsustainable energy
The International Energy Agency predicts that fossil-fuel
demand, trade flows, and greenhouse gases emissions would fol-
low their current unsustainable paths through year 2030 because
of the lack of public policy and action to control the situation.
More than 80% of the energy used comes from three types of fossil
fuels: petroleum, coal, and natural gas. About 98% of CO2 emissions
are caused by fossil fuel combustion. Fig. 3 illustrated the per-
centage of world energy consumption in year 2013 [8]. Fossils fuels
source such as petroleum, natural gas, and coal contributed to
more than 78.4% of total energy sources of the world. The
renewable energy contributes a total of 19% of world consumption,
in which 10% from modern renewables energy and 9% from tra-
ditional biomass. The modern renewable energy consists of 3.8% of
hydropower, 0.8% of biofuels, and 5.4% of others energy sources
including biomass, solar, geothermal heat, and wind energy for
power generation.
1.3. Renewable energy sources
Renewable energy plays an important role in dealing with
energy security and environment issues at global and national
levels. Renewable energy will provide cleaner power generation
and reduce the emission of greenhouse gases to atmosphere [9].
There are a few alternative energy sources such as solar energy,
wind energy, bioenergy, geothermal energy, hydropower and
ocean energy which contribute to less environmental impact than
fossil fuels [10].
Palm oil is the world's largest source of edible oil and it is
promoted to produce biodiesel in South East Asian countries
[11,12]. Biodiesel is used to substitute diesel in either pure or
blended form and it is considered an attractive alternative fuel due
to its environmental friendly characteristics, while giving a same
functional properties as diesel. Besides palm oil, rapeseed oil and
J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197
soybean are also the main sources of biodiesel. Nevertheless,
several concerns have been raised about the sustainability of this
edible oils for biodiesel production, such as competition with
agricultural land and food versus fuel fued [3].
Industrial Global biodiesel production in 2010 was around 18% of the total
Residental global biofuels production, and these productions had grown
Transport exponentially over the past decade since 2000. Germany is the
Electricity largest biodiesel producing country which contributes about 15%
of the global biodiesel production, followed by Brazil (12%), France
(11%), and Argentina (6%). Demand for biodiesel are forecasted to
2016 2020 increase in the future years due to future demand by many nations
such as the United States, European Commission, Brazil, China,
India and countries in South-East Asia such as Malaysia, Indonesia,
and Thailand [3].
In recent years, microalgae have become a focus in both aca-
demic and commercial biofuels research. Microalgae have the
potential to provide a high-yield source of biofuels without com-
Fossil Fuels promising food supplies, forest or arable land. Microalgae repre-
sent a large number of different photosynthesis species in both
Nuclear power
heterotrophic and autotrophic. The autotrophic microalgae species
Hydropower
can fix inorganic carbon from CO2, to form mainly carbohydrates,
Biofuels in which the carbohydrate can be converted into fermentable
Traditional biomass sugars for further conversion to bioethanol by selected microbial.
Others The heterotrophic microalgae species take up organic mole-
cules and convert into mainly lipids, which is used in biodiesel
production [3]. These microalgae are known to produce high oil
and biomass yields, can be cultivated with saline water and was-
tewater, can be grown on non-arable land and do not compete
with common food resources [1]. The microalgae biomass can be
converted into different kinds of biofuels using different processes
such as liquefaction, pyrolysis, gasification, transesterification,
fermentation, and anaerobic digestion. The production of biofuels
via transesterification of microalgae oil, and alcohol from micro-
algae carbohydrate via hydrolysis and fermentation are becoming
more important in the future [13].
2. Biofuels as an alternative to fossil fuels
In the recent years, many countries in the world have made
extensive effort to reduce fossil fuel energy demand and improve
energy conversion efficiency [14]. Currently, 88% of global energy
consumption is mainly derived from fossil fuels, such as petrol,
coal, and natural gas through combustion [15]. These fossil fuels
are non-renewable and will deplete overtime. Therefore, a more
promising new strategies is required for energy security purpose
as well as to mitigate CO2 emission [16].
Biofuels have seen as potential sources to meet future energy
demand. There are rapid global growth in liquid biofuels in the
past few years [16]. The first generation biofuels have reached
commercial level and already established in USA, Brazil, and the
European Union. These biofuels are mainly derived from food and
oil crops including sugarcane, sugar beet, vegetables oils and
animal fats. However impact of these biofuels in transportation
sector is still limited due to directly competing with food for crops
use and agricultural land [14,16]. The second generation biofuels
derived from biomass sources are mostly agricultural residue,
forest harvesting residue and wood processing residues and non-
edible components from food crops along with cultivation of non-
food crops such as jatropha, mahua, tobacco seed and miscanthus
[2]. Therefore they are not directly competing with arable land and
have a lower environment impact than first generation. However,
second generation biofuels have low conversion rates and the
conversion processes are not economically feasible at this moment
[2,14].
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197 183

Table 1 Table 2
Comparison of various biodiesel sources [4,17–26]. Classification of microalgae according to algae color [2,29].

Crop Seed Oil Land area Viscosity Combustion Type Color Group
oil (% yield (L use 40 °C heat (kJ/g)
oil by oil/ha (m2 year/kg (mm2/s) Microalgae Diatoms Bacillariophyceae
wt) year) biodiesel) Yellow–green algae Xanthophyceae
Cyanobacteria (blue–green algae) Cyanophyceae
1st Generation Golden algae Chrysophyceae
Corn 44 172 66 31 – Green algae Chlorophyceae
Hemp 33 363 31 – – Macroalgae Red algae Rhodophyceae
Soybean 18 446– 18 26 38.37 Green algae Chlorophyceae
636 Brown algae Phaeophyceae
Safflower 20.1 779 – 30 –
Chinese Tallow 12–45 907 – – –
Camelina 42 915 12 – – cultivation, b) harvesting, c) lipid extraction and transesterification
Sunflower 40 952– 11 – –
for biodiesel production and d) fermentation, starch hydrolysis
1070
Peanut 70 1059 – 22.7 39.9 and distillations for bioethanol production. A process flow of
Canola 41 974– 12 33 38.52 biofuels derived from microalgae biomass is shown in Fig. 4
1190 The constituents of lipids, protein and carbohydrates of 22
2nd Generation
Castor 48 1307– 9 – –
different microalgae, where lipid are extracted to produce bio-
1413 diesel whereas carbohydrates are extracted to produce ethanol,
Jatropha 20–60 1892 15 42.5 39 are shown in Table 3 [13,18,29,30,33]. The lipid content of micro-
Polanga 65–75 2000 – – –
algae is highly dependent on the microalgae species and cultiva-
Karanj 25–40 2590 – 27.82 39.1
Coconut 65–75 2689 – 29.6 37.94 tion conditions. Microalgae have the capability to grow rapidly and
Oil palm 36 5366– 2 38 38.3 could accumulate large amounts of natural lipid within their cells,
5950 which is approximately 20–70%. To make economical viable
3rd Generation
microalgae fuel for power generation, choosing appropriate
Microalgae 30–70 58.7– 0.1–0.2 36.6 –
136.9k microalgae strain is crucial [31].

Biofuel derived from microalgae are classified as third genera-
tion biofuels and have potential for large-scale production. Table 1
shows various feedstock for biofuels production. From the table,
microalgae seems to be the only possible feedstock to completely
replace fossil fuel [4,17–26]. Microalgae can grow rapidly and have
the ability to live in harsh condition. The growth of microalgae can
be accelerated through carefully selecting right species with cor-
rect environment condition. Normally, microalgae are cultivated in
large open ponds or photo-bioreactors (PBR) to produce biomass
and subsequently harvested for further processing to biofuels.
3. Biofuels production from microalgae
During the last decades, scientists have been searching for new
alternative energy sources to replace fossil fuels and they have
discovered that photosynthetic microorganism such as microalgae
have the potential to become alternative sources. Microalgae have
the ability to convert solar energy into chemical forms through
photosynthesis and have higher growth rate than plants [16,27].
Moreover they can live in diverse environment with simple
nutrient requirements. These advantages give opportunity to cul-
tivate them in areas that are not usually supported by mainstream
agricultures [27,28].
Algae are a large and diverse group of simple aquatic organism
that are mostly microscopic [28]. Microalgae are unicellular pho-
tosynthetic microorganisms that are lack of cell types found in
land plants. These microalgae convert sunlight with CO2 and water
with nutrient to produce biomass. The population of algae are
generally classified according to their color as shown in Table 2
[2,29].
Microalgae can be cultivated from various aqueous systems
such as open pond, closed pond, photo-bioreactor or hybrid photo-
bioreactors. Different algae species require different cultivation
temperatures. An optimal cultivation temperature for algae
growth should be around 20–30 °C [30]. Biofuels productions from
microalgae are complex and divided into 4 main stages: a)
3.1. Microalgae cultivation
Human have been using microalgae, such as blue–green algae,
as food sources since past thousands of years. Only in few decade
ago, the actual microalgae cultivation has begun [34]. The first ever
successfully unialgal cultured was Chlorella vulgaris by Beljerinck
in 1980. In the early 1900's, Warburg have developed plant phy-
siology study based on such culture [35]. Commercial microalgae
cultivation in large scale was started in Japan using microalgae
Chlorella species in the early 1960's and followed by cultivation
and harvesting of Athrospira species in Lake Texcoco by Sosa Tex-
coco. There are numerous research studies on microalgae but not
always resulted in commercial applications [34].
Microalgae cultivation systems are widely classified into open
or closed systems depending on their design conditions. In open
systems, microalgae are cultivated in open area environment such
as ponds, lagoons, deep channels, shallow circulating units and
others. In closed systems, microalgae are cultivated in vessels with
transparent wall and exposed under sunlight or artificial radiation
to facilitate photosynthesis [31].
Phototrophic cultivation is the most common way of micro-
algae cultivation [36]. The photosynthesis required two sources
which are solar radiation and carbon sources. The light is needed
for carbon fixation, and to increases the growth rate of microalgae
and lastly resulted in high production of microalgae biomass.
However, low intensity of light will inhibit the photosynthesis
process. The carbon source is a fundamental need for the growth
of microalgae [37–39]. The higher concentration of carbon will
result in faster growth rate and higher biomass productivity, but,
has the tendency to reduce the pH in the cultivation medium that
may inhibit the growth of microalgae [40]. Concentration of
nitrogen and carbon to nitrogen ratio are also crucial to determine
the growth rate of microalgae [41–43]. As a result, selecting right
cultivation system for microalgae biofuels production highly
depends on the type of microalgae strain, applied technology and
biological conversion process [26].
184 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197

Disposal
Natural (Industrial) Synthetic Combustion Synthetic

Water Solar energy CO2 absorption

Nutrient Microalgae culture Oxygen

Harvesting

Drying

Microalgae biomass

Biofuel Production Animal Feed Health food Combustion

(Conversion Processes) product

Biodiesel Alcohol Biogas CO2 Energy

Biofertilizer Glycerol

Fig. 4. Flow diagram of biofuels production from microalgae [31,32].

Table 3 Sunlight
Chemical compositions of microalgae in % dry matter basis [13,18,29,30,33].

Microalgae Lipids Protein Carbohydrate

Anabaena cylindrica 4–7 43–56 25–30 H2O &

Aphanizomenon flos-aquae 3 62 23 Paddle
Nutrient
Arthrospira maxima 6–7 60–71 13–16 wheel
Botryococcus braunii 86 4 20 CO2
Chlamydomonas rheinhardii 21 48 17
Chlorella ellipsoidea 84 5 16 Microalgae
Chlorella pyrenoidosa 2 57 26
Chlorella sorokiniana 20–36 21–53 5–25 Fig. 5. Open pond system (single way) [16, 29].
Chlorella vulgaris 14–22 51–58 12–17
Dunaliella salina 6 57 32
Dunaliella bioculata 8 49 4 pond system is the difficulty to control the surrounding environ-
Euglena gracilis 14–20 39–61 14–18 ment condition (e.g. temperature and weather), in which this
Prymnesium parvum 22–38 30–45 25–33 factor will directly affect the microalgae biomass productivity.
Porphyridium cruentum 9–14 28–39 40–57
Scenedesmus obliquus 12–14 50–56 10–17 Raceway pond are commonly used to cultivate Chlorella sp., Spir-
Scenedesmus quadricauda 1.9 47 – ulina platensis, Haematococcus sp. and Dunaliella salina, in which
Scenedesmus dimorphus 16–40 8–18 21–52 the highest biomass production rate are 60–100 mg/L/day of dry
Spirulina maxima 6–7 60–71 13–16
Spirogyra sp. 11–21 6–20 33–64
weight [29, 45].
Spirulina platensis 4–9 46–63 8–14 In a raceway pond system as shown in Fig. 5, the microalgae,
Synechococcus sp. 11 63 15 water and nutrition are circulated around a racetrack using paddle
Tetraselmis maculata 3 52 15
wheels [29]. This will keep the microalgae suspended in water and
allow utilization of CO2 from atmosphere. The pond is shallow to
3.1.1. Open pond systems allow light penetration into the pond in order to maximize light
There are many types of open ponds used for microalgae cul- exposure to microalgae for photosynthesis. Raceway pond oper-
tivation such as raceway, shallow ponds or circular ponds [44]. ated in a way that continues supply of water and nutrition while
Water and nutrition are supplied to microalgae by channeling mature microalgae will be removed on the other end. The benefit
runoff water from nearby land area, industrial disposal water or of this shallow pond is some CO2 waste can be reused by bubbling
water treatment plant. The most common open system is raceway back into the pond. In fact, carefully controlling other parameter
pond due to its potential to produce large quantity of microalgae such as pH and other environment conditions will allow the uti-
for commercial application. However, the problem with raceway lization of 90% injected CO2 [29,44].
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197 185

Table 4
Biomass productivity of several species microalgae in various system [16,32,46–50].

Species Production system Volume (L) Xmax (g L " 1) Pmax (g L " 1day " 1)

Anabaena sp. Open pond – 0.23 0.24

Arthrospira Tubular 146 2.37 1.15
Arthrospira platensis Undular row tubular 11 6.00 2.70
Botryococcus braunii Continuous bioreactor – 0.40 0.05
Botryococcus sp. Jar 9 – 0.04
Chlamydomonas reinhardtii Roux bottle – – 0.06
Chlorella Flat plate 400 – 3.2–3.8
Chlorella sorokiniana Inclined tubular 6 1.50 1.47
Chlorella sp. Open pond – 40.00 –
Chlorella vulgaris Jar 9 – 0.07
Chlorococcum Parabola 70 1.50 0.09
Chlorococcum Dome 130 1.50 0.10
Chlorococcum sp. Bag photo-bioreactor – – 3.83a
Chlorococcum sp. Bag photo-bioreactor – – 10b
Dunaliella tertiolecta Culture flasks 0.65 – 0.39
Haematococcus pluvialis Parallel tubular (AGM) 25,000 – 0.05
Haematococcus pluvialis Bubble column 55 1.40 0.06
Haematococcus pluvialis Airlift tubular 55 7.00 0.41
Haematococcus pluvialis Flat plate 25,000 – –
Haematococcus pluvialis Open pond – 0.20 –
Nannochloropsis sp. Flat plate 440 – 0.27
Nannochloropsis sp. Raceway 2000 0.50 –
Nannochloropsis sp. PBR 135 0.50 –
Phaeodactylum tricornutum Airlift tubular 200 – 1.20
Phaeodactylum tricornutum Airlift tubular 200 – 1.90
Phaeodactylum tricornutum Outdoor helical tubular 75 – 1.40
Porphyridium cruentum Airlift tubular 200 3.00 1.50
Scenedesmus sp. Jar 9 – 0.07
Spirulina platensis Tubular 5.5 – 0.42
Spirulina platensis Open pond – 0.47 0.05
Spirulina platensis Conventional Flask type 1 – 0.08
Spirulina platensis Flat type 1 – 0.16
Tetraselmis Column – 1.70 0.42

Xmax maximum cell concentration (g/L).

Pmax maximum oil production (g/L/day).
a
Biofuel produced is ethanol (g/L).
b
Biofuel produced is biodiesel (g/L).

3.1.2. Photo-bioreactors (PBR)

Photo-bioreactors technologies are designed to overcome the
problems associated with open pond system. Photo-bioreactors
will allow the cultivating of single species microalgae for longer
period than open raceways and able to produce large quantity of
microalgae biomass [17,30,44]. There are many types of closed
system including the column, tubular and the flat plate photo-
bioreactors. These systems are more practical and productive in
order to cultivate microalgae due to the cultivation conditions can
be effectively controlled (pH, temperature, CO2 concentration and
etc.) and reduce the risk of contamination. Several microalgae
species with their respective biomass productivity in various
photo-bioreactors are shown in Table 4 [16,32,46–50]. However,
capital costs of closed system are much higher than open systems
[17].
The basic design of photo-bioreactors (as shown in Fig. 6)
consists of array of straight glass or plastics tubes. The tabular
array can be rearranged in horizontal, vertical, inclined or a helix
position to allow optimum exposure to sunlight. The earliest
design of closed system is flat-plate and often used in research
Fig. 6. Basic design on airlift photo-bioreactor [16].
studies due to its larger surface area and maximizing solar energy
absorption by microalgae [16,44].
nutrient rich open pond system to facilitate biomass production.
3.1.3. Hybrid photo-bioreactors For example, the hybrid systems are implemented on Haemato-
The hybrid systems are the combination of closed photo- coccus pluvialis with annual production rates of 10–24 toe per ha.
bioreactors and open system for different growth stages. The Hence, using higher lipid content of microalgae supplied with
first stage is to cultivate the microalgae in closed photo- appropriate nutrients and at controlled temperature and CO2
bioreactors in order to minimize contamination from environ- concentration will result in massive production of microalgae
ments whereas the second stage is to expose the microalgae in biomass [16].
186 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197

Table 5
Advantages and limitations of open and closed systems [16,17,29,31,35,45,48,51,52].

Production system Advantages Limitations

Raceway pond Low capital cost Poor biomass productivity

Low operation cost Large area of land are needed
Easy to clean Limited to a few strains of microalgae
Utilizes non-agricultural land Poor light and CO2 utilization efficiency
Low energy inputs
Easy maintenance Cultures are easily contaminated
Simple system Evaporation and water losses are significant
Easy to scale up for commercial level No cooling system
Difficulties in growth controlling
Harvesting cost are relatively higher
Poor mixing efficiency
More than one species can be cultivated

Tubular Suitable for outdoor cultures High capital cost

photobioreactor Large illumination surface area Requires large land space
Large quantity of microalgae biomass Gradients of pH, dissolved O2 and CO2 along the tubes
Single species culture Required supplied of air to operate by using airlift pumps
Less than 0.1 m diameter to ease penetration of light High concentration of O2 will inhibit photosynthesis
Easy to design, install and operate
Biomass recovery cost are much lower due to high biomass Limits the maximum length of continuous run tube
concentration Some degree of wall growth
Shorter harvest time Fouling
Higher surface to volume ratio

Flat plate Relatively cheap Difficult large scale-up

photobioreactor High biomass productivities Difficulties in controlling temperature
Easy to clean and sterilize Limited degree of growth at near wall region
Low oxygen accumulation
Large illumination surface area Difficulties in controlling carbon dioxide diffusion rate
Good light path
Low contamination of microalgae Microalgae strains may occur
Good for microalgae immobilization Photo-inhibition may occurs
Hydrodynamic stress may occur
Good for outdoor and indoor cultures

Column Compact Relatively expensive

photobioreactor High mass transfer High maintenance cost
Low energy consumption Small illumination area
Good mixing with low shear stress Higher shear stress on microalgae cultures
Easy to sterilize
Easy to work under sterile Sophisticated construction material
conditions Hard to scale-up
Good for microalgae immobilization Complex flow pattern
Low surface and volume ratio
Less photo-inhibition and photo-oxidation problem Large unit in order to build due to limited adjustable in diameter and
height

The pros and cons of various cultivation systems are tabulated
in Table 5 [16,17,29,31,35,45,48,51,52]. By comparing open pond
system with closed photo-bioreactor, open pond systems are much
cheaper for large-scale production, with low requirement of
energy input. Averagely, the open pond is built with capacity
ranging from 2000–5000 m2, where the largest raceway pond
occupied about 440,000 m2 [29]. However, open pond systems are
less efficient than closed photo-bioreactor in term of biomass
productivity and cultivation time. This will result in lower rate of
profit return and longer payback period for open pond system.
3.2. Harvesting
Water removal is required for wet paste microalgae for sub-
sequent processing of biofuels. However, harvesting process is
usually costly and accounted about 20–30% of total biofuels pro-
duction cost [29,53]. Therefore, it is important to choose appro-
priate harvesting methods to minimize the overall cost. The har-
vesting methods are highly dependent on the type of microalgae,
cells density and desired target products [16,53].
Harvesting of microalgae can be divided into two step process.
The first step is bulk harvesting where biomass is separated from
the bulk culture, such as flocculation, flotation or sedimentation
[16]. The second step is to concentrate the biomass slurry via
techniques like centrifugation and filtration. These two processes
are called thickening [16,53].
3.2.1. Flocculation
Normally, microalgae cells carry negative charge that does not
allow self-aggregate in suspension. With the chemical called
flocculants such as multivalent cations and cationic polymers, the
flocculants will neutralize the negative charge in microalgae that
caused them to aggregate (floc) and fasten the sedimentation rate.
Flocculants that commonly used are ferric chloride, ferric sulfate,
aluminum sulfate and other organic salts. [16,53]. Khoo et al. used
ferric chloride at rate of 125 mg/l to process 2000 l of Nanno-
chloropsis sp. and the process consumed 16.7 kJ of energy [48].
Mature microalgae that continuously exposed under sunlight with
minor supply of CO2 will cause autoflocculation in which this
method is more favorable for large scale harvesting as the cost of
harvesting will be reduced significantly compared to conventional
flocculation method [51].
3.2.2. Filtration
The filtration process involved the microalgae culture runs via
filters by trapping microalgae and only allow water to pass
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197
through the filter. There are many types of filtration, such as
microfiltration, dead end filtration, vacuum filtration, pressure
filtration, ultrafiltration and tangential flow filtration (TFF) [53].
The conventional filtration and vacuum filtration are suitable
for large microalgae with size more than 70 mm such as Spirulina
and Coelastrum, but not suitable for microalgae where their size is
less than 30 mm such as Scenedesmus, Chlorella and Dunaliella.
Microalgae with smaller size are recommended to use membrane
microfiltration and ultrafiltration, in which both filtration methods
required low trans-membrane pressure and low cross flow velo-
city conditions. Microfiltration membrane has pore sizes between
0.1 and 1 μm whereas ultrafiltration membrane has smaller pore
size between 0.001 and 0.1 μm [16].
TFF method are known with its high rate filtration and it is
applied to a wide range of biological fields, as it can be used to
concentrate and desalt sample solution in various scale size and it
is used to separate solutions containing viruses, bacteria or cellular
material. As TFF is used in filtration of microalgae, it proved that
approximately 70–80% of microalgae are able to recover by using
this method while maintaining the structure, properties and
motility of the filtered microalgae. The main drawback of TFF is the
cost required to replace the membrane is very high and it needs to
be replaced very frequently when used on a larger scale of
production.
3.2.3. Gravity and centrifugal sedimentation
The gravity sedimentation is the most common techniques
used to harvest microalgae biomass especially in wastewater
treatment. This method is suitable for microalgae with high den-
sity and large size such as Spirulina, [16,53] Botryococcus sp., C.
vulgaris, and Scenedesmus sp. [47].
Centrifugal method involves centripetal acceleration to sepa-
rate the microalgae culture into two different density areas –
greater and less density areas. The process is rapid and energy
intensive which need high energy cost and high maintenance cost
as it involves free moving parts. The harvesting efficiency is more
than 95% and slurry concentration can be increased by up to 150
times for 15% of suspended solids [16,53].
3.3. Extraction and purification for microalgae biomass
Dehydration process is an immediate process after harvesting
which is important for dried microalgae biomass production.
Drying methods that are usually used are sun drying, low pressure
shelf drying, spraying, drum drying, fluidised bed drying, freeze
drying (lyophillized) and refractive windows technology drying.
Expenditure of sun drying is the cheapest among all dehydration
methods, but require large surfaces area and very time consuming.
Other methods are more efficient but the cost of dehydration are
very expensive especially for large scale operations [16]. Johnson
and Wen and Kobayashi et al. both used freeze drying method to
dehydrate biomass of Schizochytrium limacinum and Chlorella sor-
okiniana [33,54]
There are various types of lipids and fatty acid produced from
microalgae which can be extracted for biofuels production [55].
The lipid extraction method and efficiency are highly dependent
on microalgae species and their chemical composition [16]. The
methods that usually used to extract microalgae lipid are physical,
chemical with organic solvent, and super critical methods. The
most practicable approach used in biofuels production is through
solvent extraction [55].
3.4. Conversion process and biofuels production
Biomass produced from microalgae can be converted into
biofuels through a number of ways. The conversion of biomass into
187
biofuels can be classified into three sub classes – chemical con-
version, biochemical conversion and thermochemical conversion
which are shown in Fig. 7 [2,26,31,52]. Selecting appropriate
conversion methods is determined by quantity and type of bio-
mass feedback, desired final energy form and economic viability
[16].
3.4.1. Chemical conversion
3.4.1.1. Transesterification. In biodiesel production, processes of
chemical conversion from microalgae biomass to biodiesel occur
mainly via transesterification. The process of triglyceride (lipid)
reacting with a mono-alcohol in the presence of acidic, alkaline or
enzymes based catalyst to produce mixture of fatty acid methyl
ester (FAME) and glycerol is shown in Fig. 8 [31].
There have been several studies on biofuels from microalgae
via transesterification. One study reported that using alkaline
catalyst in transesterification of oil from microalgae Spirogyras and
Oedigonium, 90% of conversion of biodiesel can be attained. There
are two ways to perform transesterification, which are conven-
tional (two-stage methods) and direct transesterification (single-
stage method). By using conventional transesterification, dried
biomass will go through lipid extraction process via mechanical or
chemical methods, before diverted to transesterification and pur-
ification steps. The other method which is known as in-situ or
direct transesterification involved lipid extraction and transester-
ification simultaneously as shown in Fig. 9 [54,56,57]. Johnson and
Wen have used both methods to produce biodiesel from micro-
algae S. limacinum; conventional transesterification method
resulted in 57% of crude biodiesel (% of dry biomass) and FAME of
66.37% while chloroform based direct transesterification method
resulted in 66% of crude biodiesel (% of dry biomass) and FAME of
63.47% [54].
3.4.1.2. Esterification. The esterification process is a reversible
reaction process. The reaction converts free fatty acids to alkyl
ester through acid catalyst. The goal is to reduce formation foam
and improve conversion of biodiesel. Monoraphidium contortum
microalgae were used as a feedstock for microalgae lipid ester-
ification and the result found that the conversion into methyl
esters is averagely 91.7% [31].
3.4.2. Thermochemical conversion
Thermochemical conversion is a process of thermal decom-
position of biomass into fuels products. Such conversions are
gasification, pyrolysis and liquefaction [46]. The list of thermo-
chemical conversion for microalgae, oil yields and their calorific
value are shown in Table 6 [16,46]. Thermochemical conversion
has several advantages such as smaller carbon footprint, better
nutrition recovery, short processing time, no emission of fugitive
gas, work well with either wet or dry biomass, capable in handling
various feedstock and blends, pathogens and pharmaceutical
active compound are eliminated due to high temperature and
minor residue after conversion (e.g. ash) [31].
3.4.2.1. Gasification. Gasification is a thermochemical process in
which microalgae biomass is partially combusted to synthesis gas
(syngas) at high temperature ranging from 800 to 1000 °C with
appropriate pressure [52,53]. The syngas has low caloric value and
can be directly used as fuel for engines and turbines [31]. Tsuka-
hara et al. used a low temperature catalytic gasification with
nitrogen cycling at a temperature of 400 °C and pressure of
20 MPa to produce ammonia, methane gas, and carbon oxides
from microalgae biomass. Ammonia is then supplied to the
microalgae as nutrient for cultivation process. The whole process is
illustrated in Fig. 10 [46,58].
188 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197

Transesterification Biodiesel
Chemical
Conversion
Esterification

Fuel Gas
Microalgae Gasification
biomass
Bio-Oil
Liquefaction Gases
Solid Residue
Thermochemical
Conversion Pyrolysis
Bio-Oil
Hydrogen
Direct
Combustion

Anaerobic Methane
Biochemical Digestion Hydrogen
Conversion

Alcoholic Alcohol
Fermentation Ketone

Interesterification Biodiesel

Fig. 7. Conversion process of microalgae biomass [2,26,31,52].

O O
CH2 —C—R1 CH3 — O—C—R1 CH2 —OH
O O
Catalyst
CH —C—R2 + 3CH3—OH CH3 — O—C—R2 + CH —OH
O O
CH2 —C—R3 CH3 — O—C—R3 CH2 —OH

Triglyceride Methanol Methyl ester Glycerin

Fig. 8. Triglyceride to glycerol via transesterification [31].

Conventional Transesterification (two stage method) Glycerol &Water

Dried Lipid
Transesterification Purification Biodiesel
microalgae extraction

Direct Transesterification (single stage method)

Dried Direct Purification Biodiesel

microalgae Transesterification

Glycerol & Water

Fig. 9. Conventional transesterification and direct transesterification method [54,56,57].

 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197 189

Table 6 content moisture content of microalgae biomass required extra

Comparison of thermochemical conversion for microalgae [16,46]. cost to dry [16].
Conversion Microalgae Temperature Liquid
process 3.4.3. Biochemical conversion
(°C) Oil Calorific Biochemical conversion is a process of converting biomass into
yield Value others fuel through anaerobic digestion, alcoholic fermentation
(% dry (MJ kg " 1) and interesterification [46].
wt.)

Liquefaction Botryococcus braunii 300 64 45.9 3.4.3.1. Anaerobic digestion. Anaerobic digestion is the process of
Liquefaction Dunaliella tertiolecta 300 42 34.9 converting organic waste into biogas gaseous. This process mainly
Liquefaction Dunaliella tertiolecta 300 37 36 involved methane and carbon dioxide production, in which the
Pyrolysis Chlorella 450 57.9 41
gaseous produced can be used directly as cooking fuel, generating
prothothecoides
Pyrolysis Chlorella 450 16.6 30 power gas engines, or upgraded into natural gas-quality bio-
prothothecoides methane. Anaerobic digestion is suitable for all organic materials,
Pyrolysis Chlorella 500 18 30 and selection of digestion system depends on moisture content of
prothothecoides
microalgae and the carbon (C) and nitrogen (N) ratio of the
Pyrolysis Chlorella 502 55.3 39.7
prothothecoides
microalgae in which, the optimal C and N ratio is 20–30:1, as
Pyrolysis Microcystis 500 24 29 excessive of N will lead to ammonia inhibition of digestion [16,31].
aeruginosa Anaerobic digestion is a process of breaking down organic
material by bacterial hydrolysis, subsequently the acidogenic
bacteria will convert the sugar and amino acids into carbon
dioxide, hydrogen, and ammonia, whereas the methanogens
Microalgae cultivation converts the intermediate products into end products which are
methane and carbon dioxide [16,31]. Kobayashi et al. used C. sor-
okiniana with 10% anaerobic digester effluent from cattle manure
Harvested microalgae (wet)
digestion. The result shows that the highest accumulation of
protein and starch exhibited was 34% and 23% of ash free dry
Low temperature catalytic gasification weight respectively but lower production of lipid. Currently,
anaerobic digestions are commonly used in macroalgae species
such as Macrocytis pylifera, Sargassum and Laminaria in order to
Methane and Hydrogen CO2 and ammonia produce biogas [33,54].

3.4.3.2. Alcoholic fermentation. Fermentation is a process of con-

Energy
Fig. 10. Flow diagram of biomass low temperature catalytic gasification [46,58].
3.4.2.2. Thermochemical liquefaction. Thermochemical liquefaction
is a process of converting wet microalgae biomass materials to
biofuels. The process involves temperature between 200 and
350 °C and at high pressure between 5 and 20 MPa with catalyst
used to yield bio-oil and hydrogen. The advantage of this process is
their ability to convert wet biomass into energy, but initial cost is
high due to complex design of fuel-feed system and reactors [16].
3.4.2.3. Pyrolysis. Pyrolysis is a process to convert biomass in the
absence of air/oxygen into bio-oil, charcoal and gaseous products
at temperature between 350 and 550 °C [59]. Pyrolysis process can
be further classified into three classes: conventional pyrolysis, fast
pyrolysis and flash pyrolysis. Biomass pyrolysis have the potential
for large-scale biofuels production, especially the fast pyrolysis
process which is feasible and effective to produce high quality and
high yield fuel oils [60]. Comparing various conversion technolo-
gies in Table 6, pyrolysis process is reliable and promising for
commercial biofuels production from microalgae [16].
3.4.2.4. Direct combustion. Direct combustion process is burning of
biomass in air to convert the chemical energy stored in biomass
into hot gases at temperature around 800–1000 °C. In practice,
combustion biomass burnt at a moisture content level less than
50% unless pre-dried. The combustion plant output ranges from
100 to 3000 MW depending on the scale of industrial plants [61].
However, direct combustion of biomass required pre-treatment
such as drying, chopping and grinding. Direct combustion of bio-
mass in large plants has advantages over conventional coal-fired
plants due to better conversion efficiency. However, the high
verting sugar, starch or cellulose contain inside biomass materials
into bioethanol. The microalgae biomass is ground down and the
starch is hydrolyzed to sugar by enzymes before diverting to yeast
fermentation to produce bioethanol. The ethanol is purified by
distillation process to remove the water and impurities [61].
Microalgae with high starch content such as C. vulgaris is a good
source of bioethanol and up to 65% bioethanol conversion could be
attained [16].
3.4.3.3. Interesterification. Interesterification reaction is a process
of using enzyme as catalyst to produce fatty acid esters. This can
be achieved by using high activity enzyme such as Candida Ant-
arctica, which would produce triacetin and long-chain fatty acid
methyl or ethyl ester. In the recent years, there is a growing
interest of interesterification used in biodiesel production where it
yields highly pure product and the byproducts can be easily
removed. However, the cost of enzyme is high which make the
process not economically feasible at the current stage [31].
4. Energy generation from microalgae biofuels
Biomass and biofuels derived from microalgae can be used in
power production. Microalgae biofuels has high potential to
replace fossil fuel for internal combustion engine. Electricity can
be generated using microalgae in coal co-firing to mitigate
greenhouse emission and reduce coal consumption.
4.1. Microalgae biofuels for internal combustion engine
In recent years, production of biofuels from renewable feed-
stock for transportation has become a promising approach [62].
There are two main fossil fuels used in transportation, which are
190 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197
Transportation Fuels
Biorenewable Fuels Petroleum Fuels
Bioethanol Bio-hydrogen Bio-diesel
Gasoline Diesel
Bio-oil Fuel Cells
Otto Engine Diesel Engine
Fig. 11. Renewable fuels for transportation [62].
Table 7
Standard specification for fuel properties [55].
Property ASTM D6751 EN 14214
Cetane number 47 min 51 min
Cold flow Cloud point: Cold filter plugging point: varies
report by location and time of year
Kinematic viscosity; 40 °C; 1.9–6.0 3.5–5.0
mm2 s " 1
Oxidative stability; h 3 min 6 min
gasoline and diesel fuel. These fossil fuel can be replaced with
biofuels derived from microalgae biomass (bioethanol and bio-
diesel). Fig. 11 shows the diagram of renewable fuels and petro-
leum fuels used in transportation [62].
The possibility of microalgae biofuels to replace biodiesel pro-
duction is ultimately determined by its chemical properties. These
chemical properties depend on fatty acid profile. There are two
biodiesel standard in the world served as guideline and assess-
ment of biodiesel quality. The standards are ASTM D6751 in US
and EN14214 in Europe (Table 7). The cetane number indicate the
ignition and combustion characteristics of fuels [55].
The fatty acid methyl esters profile of microalgae is depending
on the microalgae growth parameter such as environment tem-
perature, light intensity, nutrition and time of growth [54]. Fatty
acid compositions of several microalgae species are shown in
Table 8 [47,49,54,63–68]. The high content of oleic acids (C18:1)
have better balance of fuel properties including ignition quality,
combustion heat, cold filter plugging point, oxidation stability,
viscosity and lubricity. The higher oleic acid content in fatty acid of
microalgae give better oxidation stability for long storage and
reduce in cold filter plugging point used in cold region. Therefore,
higher oleic acids content in fatty acid of microalgae species are
most suitable for biodiesel production [47,54,63]. Mixed culture of
Chlorella sp. and Saccharomyces cerevisiae, Picochlorum sp.,
Botryococcus sp., Scenedesmus sp. and Nannochlopsis oculata con-
tains large amount of oleic acid, which make them suitable for
biodiesel production.
Biofuels such as methanol, ethanol and biodiesel are oxyge-
nated compounds that produce cleaner emission in combustion
process. Compared to fossil fuels, biofuels provide better com-
bustion performance. Many studies have shown that fossil fuel
blended with biofuels or pure biodiesel gives better performance
in internal combustion engine [62].
4.2. Electricity production by biomass from microalgae
Large amount of electricity generated via coal fired power plant
in the world has significant impact towards environment. Con-
sumption of coal in the world are expected to increase about 56%
from 2007 to 2035 [69]. An alternative to coal is biomass, which is
considered as an attractive renewable fuel. Among all the biomass
feedstock, microalgae species cultivated in open pond system is
considered the most promising source for co-firing [70].
Utilizing biomass for Co-firing is a cost-effective approach to
generate electricity. Originally this approach is meant to reduce
greenhouse gas emission by fossil fuels, trace metal and to provide
a broader base of biofuels supplies in order to support the devel-
opment of infrastructure for fuel supply and delivery [71]. Fig.12
shows the comparison of two scenarios for electricity generation
in power plant – conventional coal firing and coal/microalgae co-
firing [46,72]. By cultivating microalgae using CO2 produced from
power plant can substantially reduce emission of CO2 to atmo-
sphere, and co-firing the biomass from microalgae can generate
additional electricity.
Replacing coal with microalgae biomass in power plant can
subsequently reduce the greenhouse gas emission. However dur-
ing the microalgae production there will be input of other energy
and fertilizer which are equally important as it results in high
depletion of natural resources and eutrophication. The eutrophi-
cation is high due to the fertilizer consumption during the pro-
duction of microalgae. Therefore, the balance between the pro-
duction of microalgae and the microalgae co-firing need to be met
in order for the process to be more environmental feasible [72].
5. Co-processes in microalgae cultivation
5.1. Potential mitigation of CO2 emission by microalgae
Greenhouse gaseous has caused global warming to a point that
cannot be ignored. There are many developments in technologies
to reduce greenhouse in atmosphere, mainly conservation, direct
mitigation and indirect mitigation. Conservation method is to
reduce electricity consumptions, direct mitigation is to capture
CO2 directly from atmosphere and indirect mitigation is to offset
CO2 by reducing CO2 emission from production [73]. CO2 mitiga-
tion by microalgae biofuels production is considered as indirect
mitigation method.
The importance of biological CO2 mitigation has received much
attention recently especially in microalgae cultivation since CO2 is
an essential carbon source for microalgae growth and indirectly
used as an alternative way to mitigate CO2 [74]. However the rate
of carbon uptake by microalgae is limited by photosynthesis pro-
cess and it is crucial to understand microalgae metabolism before
cultivating microalgae in a large scale [70]. Microalgae has the
advantage in capturing CO2 compared to other land-based plant as
its effectiveness of photosynthesis conversion are high and resul-
ted in high production of microalgae biomass. Microalgae is cap-
able to produce different types of biofuels and its ability to adapt
and grow in harsh environment with CO2 fixation from atmo-
sphere or flue gas [71] and biogas (60% methane and 40% CO2)
[75].
Taher et al. have conducted a study on Chlorella sp., Pseudo-
chlorococcum sp. and Nannochlorpsis sp. to determine the optimum
conditions for lipid productivity and CO2 fixation. The microalgae
are cultured in both nitrogen sufficient and nitrogen starvation
under CO2 concentration ranging from 0.04% to 2% (v/v) and var-
ious salinities levels. The Chlorella sp. has the highest CO2 fixation
rate of 1.63 g L " 1d " 1, followed by Pseudochlorococcum sp. with
1.24 gL " 1d " 1 and Nannochlorpsis sp. with 0.55 gL " 1d " 1[65].
5.1.1. Metabolism and photosynthesis of microalgae
Microalgae are usually unicellular. In prokaryotic microalgae
cell, a region called chloroplast, contains photosynthetic mem-
branes. For eukaryotic microalgae usually divided into different
light harvesting – Rhodophyta, Chrysophyceae, Phaephycease and
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197 191

Table 8
Fatty acid composition of several microalgae species (g/100 g of fatty acid) [47,49,54,63–68].

Fatty acid C14:0 C16:0 C16:1 C16:2 C16:3 C16:4 C18:0 C18:1 C18:2 C18:3 C20:0 C20:2 C20:5 C22:6

Amphidinium sp. 0.8 35.7 1.1 – – – 4.1 19.4 – – 5.7 11.9 20.2 –
Bidulphia sp. 21.4 23.6 33.2 1.7 4.4 – 0.8 2.1 – – – 9.6 – –
Botryococcus sp. – – 4.8 – – – 4.3 55.7 34.2 0.2 – – – –
Chlorella pyrenoidosaa 0.2 34.2 5.24 0.6 – – 4.6 35.9 18.8 – – – – –
Chlorella vulgaris – – – – – – 3.4 16.3 79.4 0.1 – – – –
Chlorococcum sp.b 0.6 19.1 4.1 2.6 – – 2.2 62.8 3.8 – – – 1.4 –
Dunaliella tertiolecta – 27.1 0.7 1.3 1.0 – 0.6 16.2 11.1 45.5 – – – –
Extubocellulus sp. 6.6 25.6 60.6 2.7 – – 0.9 3.2 – – – – – –
Franceia sp. 0.6 12.9 7.3 2.3 2.0 17.0 0.5 15.2 1.2 32.3 – 0.9 – –
Mesotaenium sp. 0.5 13.4 6.1 2.9 2.4 16.4 0.6 19.4 1.0 30.4 – 0.8 – –
Nannochlorpsis oculata 5.8 32.2 29.6 – – – 1.0 21.4 – – – 8.3 – –
Nannochlorpsis sp. 6.8 31.6 – – – – 3.5 34.5 9.2 – 0.2 – – 0.65c
Phaeodactylum tricornutum 3.3 23.6 48.2 1.4 4.3 – 0.8 5.9 – – – 12.1 – –
Picochlorum sp. 0.5 16.8 1.2 5.1 3.5 – 3.4 51.3 14.9 – 2.1 – – –
Scenedesmus dimorphos 0.5 15.8 5.2 2.9 2.1 15.6 0.6 21.7 1.0 25.0 – – – –
Scenedesmus obliquus 1.1 20.3 – – – – 5.0 28.9 30.5 14.2 – – – –
Scenedesmus obliquusd 0.2 26.4 3.7 1.4 3.6 57.3 6.3 – – – – –
Scenedesmus sp. – – – – – – 0.7 57.2 36.8 2.2 – – – –
Schizochytrium limacinum 5.3 56.8 – – – – – – – – – – – 29.8

a
Nitrate absence is best cultivation condition for Chlorella pyrenoidosa.
b
Total extraction time is 80 min at temperature 80 °C.
c
Value for C22:0 instead of C22:6.
d
0.3 g L " 1 nitrate concentration is the best cultivation condition for Scenedesmus obliquus.

c) Regeneration. CO2 fixate by regenerated Ribulose-P in complex

series of reaction.
d) Production. The final end product of photosynthesis are car-
bohydrates, fatty acid, and acid amino and organic acid.

There are more than 4000 studies done on carbon fixation by

microalgae. The most common studied species are Spirulina,
Chlorella, Dunaliella and Haematococcus. Botryococcus also largely
studied for hydrocarbons production. Table 9 shows the CO2
fixation rate and biomass productivity of different microalgae
species [16,31,50,51].

5.1.1.1. Chlorella vulgaris. Chlorella is a unicellular eukaryotic green

Fig. 12. Comparison of two scenarios for electricity generation in power plant –
conventional coal firing and coal/microalgae co-firing [46,72].
Chlorophyta. The major pigments that involved in absorbing light
energy are clorophylls, carotenoids and phycobilinsm. Microalgae
photosynthesis processes are much simpler compare to higher
plant and more efficient to convert solar energy to chemical
energy. There are two stages involved in converting light energy to
sugar, which are light reaction and dark reactions [76].
Plant and microalgae follow a pathway known as Calvin cycle
which is well-known as CO2 fixation pathway. Only one enzyme is
responsible for CO2 fixation that is ribulose 1.5-bisphosphate car-
oxxylase/oxygenase (Rubisco). The Calvin cycle consist 13 enzy-
matic reaction which is shown in Fig. 13. One molecule ribulose 1,
5-bisphosphate reacts with CO2 and H2O produce two molecules
of 3-phosphoglycerate [77]. CO2 fixation happened in four phases
which are carboxylation, reduction, regeneration and production
[78].
a) Carboxylation. A reaction which involves catalyst enzyme
Rubisco where CO2 react with five carbon sugar ribulose
bisphosphate to form two molecule of phosphogycerate.
b) Reduction. Energy added into ATP and NADPH2 in two steps
by phosphorylation of Glycerate-P to form diphosphoglycerate
and reduction of Glycerate-P to phosphoglyceraldehyde by
NADPH2.
algae. Chlorella has a thick cell wall which is about 100–200 nm.
This feature allowed Chlorella to produce oil even under extreme
environment such as high irradiance, high temperature and
nutrients deficiency. The cell wall serves as a protection [57,64].
Microalgae growth rate and the protein of microalgae affected
by the amount of nitrogen and carbon sources [79]. C. Vulgaris
uptake carbon via enzyme carbonic anhydrase as catalyst for
reaction hydration of CO2 to form HCO3 and proton. The estima-
tion of rate of carbon dioxide fixation was 31.8 mg CO2 L " 1 d " 1.
The optimum concentration of CO2 supply for Chlorella growth is
about 6% [57].
5.1.1.2. Spirulina platensis. S. platensis are cyanobacteria belong to
two separate generations – Spirulina and Arthrospira. They are
good in CO2 biofixation due to their high growth rate and produce
nutraceutical product C-phycocyanin (C-PC) [50]. There are 15
species in S. platensis that usually reproduce via binary fusion.
Pigment involve in photosynthetic are chlorophyll and phycocya-
nin [57]. Generally phycobiliprotien present in Spirulina acts as a
light harvesting pigments to capture wavelength that are not able
to utilize well by chlorophyll [50]. Spirulina can be found in diverse
environment such as brackish water, marches, seawater, thermal
springs and freshwater. The optimum medium for Spirulina to
grow is alkaline water with high pH of 8.5–11.0 [57]. Light inten-
sity and CO2 are very important factor to promote Spirulina to
grow. Nitrate is also an essential nutrients to support the growth
of Spirulina and affect their biochemical composition [50]. Spir-
ulina has high protein content, which is about 55–70% in dry mass
192 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197

Fig. 13. Calvin cycle [77].

analysis and good source of vitamins (B1, B2, B3, B6, B9, B12, C, D,
and E), and minerals such as calcium, chromium, copper, magne-
sium, manganese, zinc, sodium, selenium and phosphorus. They
have high amount of fatty acid [57].
5.1.1.3. Botryococcus braunii. Botryococcus braunii is a green
microalgae and cosmopolitan species. They can live in both tem-
perate and tropical fresh water and also brackish water [25]. They
are classified into three groups depend on their hydrocarbon
content-A, B, and L races [57]. Races A contain C25 to C31 odd-
numbered n-alkadienes and alkatrienes; Race B produce two types
triterpenes and L race yield single lycopadiene [25,57]. There are
many studies done on B. braunii to increase their hydrocarbon
content for biofuels production via physical optimization in 16
strains of B. braunii (A, B and L races) [25]. Hydrocarbon content is
extracted from B. Braunii through lipid extraction [25]. Conversion
from solar energy to hydrocarbon required about 3% solar energy
by B. Braunii. Since B. Braunii synthesized hydrocarbon through
photosynthesis mechanism, burning of hydrocarbon from micro-
algae does not contribute to accumulation of CO2 in the atmo-
sphere [57]. According to Baba et al. [80], light quality affect the
growth, pigment composition, colony shape and rate of CO2 fixa-
tion of B. Braunii. The most effective light source for biofuels
production is red light [80]. Conti et al. have cultivated two strains,
Kossou-4 and Overjuyo-3 at 500 L scale for 60 days with up to 98%
recovery of microalgae biomass by using bio-flocculation with
Aspergillus fumigatus [81]. B. Braunii has the ability to accumulate
high levels of hydrocarbon, which is about 15–35% in dry weight
with up to 76% of dry weight of the cell material are combustible
that accurately estimated by thermogravimetric analysis (TGA)
[25].
5.1.1.4. Dunaliella sp. Dunaliella sp. is a green alga with biflagellate
unicellular cell [57]. Dunaliella sp. grows in saltwater, wastewater
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197 193

or brackish and utilize the inorganic nutrients contain in the important in pigmentation source in aquaculture and poultry
medium. They are highly tolerance to salt, temperature and light. industries [82].
The important parameter involve in the growth of Dunaliella is
light, CO2 concentration, and photoperiod. However, the light 5.2. Water treatment potential of microalgae
source and intensity have no major effect on the production of
fatty acid in this species [49]. Dunaliella sp. is very important in Microalgae has been used as bioremediation in wastewater
industrial process due to wide variety products can be produced treatment and already applied in the past 40 years. Many studies
such as nutritional supplements, aquaculture feeds, cosmetics, have been carried out on the use of microalgae for removal of
nutrients such as nitrogen and phosphorus [83]. Microalgae are
pharmaceuticals and food coloring [57].
extensively used to remove pathogens, heavy metal and other
contamination [16, 31]. Microalgae cultivation using fertilizers
5.1.1.5. Haematococcus. Haematococcus is a chlorophyta green have negative impact on the cost of biofuels production. About
microalgae with single cell. They are capable of synthesizing and 50% of the energy used in cultivation of microalgae and CO2
producing astaxanthin pigment which is up to 1.5–6% dry weight. emission are related to fertilizer [57]. Combining microalgae bio-
The problems with this microalgae when cultivating in large scale fuels production with wastewater treatment offer a cost effective
are easily contaminated and slow growth rate [57]. This micro- strategy since microalgae can serve to improve water quality and
algae biomass concentration is less than 10 g L " 1, which means produce biomass [57]. Table 10 shows the several microalgae
99% of the culture broth is water than microalgae biomass. species used in wastewater treatment [16,31,83].
A study by Wang et al. investigated the effectiveness of green
Therefore, the water in the culture broth can be recycled for next
microalgae Chlorella cultivating in wastewater samples (collected
cultivation and the process are very costly. Haematococcus is very
from different process location zones) and demonstrated the
microalgae can significantly remove ammonia or nitrate. The lipid
Table 9
CO2 fixation rate and biomass productivity of different microalgae species composition of Chlorella was also found suitable for biodiesel
[16,31,50,51]. production. In addition, high rate microalgae pond (HRAP) can
increase photosynthetic efficiency in microalgae growth. HRAP are
Microalgae strain CO2 con- Biomass pro- CO2 consumption
shallow and their construction is similar to raceway ponds. Fig. 14
tent in the ductivity (mg/L/ rate (mg/L/d)
feed (%) d) shows the combination process flow of wastewater treatment,
microalgae culturing and HRAP [76,83,84].
Aphanothece microscopica 15 770 1440 Besides cultivation of microalgae in wastewater treatment, the
Nageli
sludge that produced at wastewater treatment plants combine
Botryococcus braunii 5 – 497
Chlorella 5 – 251.6 with integrated processes through anaerobic digestion and anae-
Chlorella kessleri 18 87 163a robic co-digestion can be used to produce methane gases and
Chlorella littoralle 40 – 1000 estimated to be able to reduce the solid in the wastewater plant
Chlorella vulgaris air 40 75.2a [85]. Microalgae such as Scendesmus obliquus [86,87], Chlorella
Chlorella vulgaris 15 – 624
Chlorella vulgaris 15 228 460.8
[86], C. Vulgaris [87], Chlorella sp. and S. platensis [88] were suc-
Dunaliella 3 170 319.6a cessfully used in wastewater treatment and its range of methane
Dunaliella tertiolecta 5 – 272.4 yield were in between 143 and 350 CH4/kg VS (various substrate).
Haematococcus pluvialis 16–34 76 143 The anaerobic co-digestion in wastewater treatment with addition
Microcystis aerusinosa 15 243 489.6
Microcystis ichthyoblabe 15 258 520.8
of carbon-rich feedstock such as wastepaper, sludge, organic solid
Scenedesmus obliquus Air 9-16 16–31 waste, food industry waste, and agricultural residues in order to
Scenedesmus obliquus 18 140 260 maintain high C/N ratio have doubled the yield of the methane gas
Scenedesmus sp. 15 304 612 production rather than anaerobic digestion alone [86].
Spirulina platensis 3 820 1580
Spirulina platensis 2.5 995 1650
Coagulation–flocculation and sedimentation with natural floc-
Spirulina platensis 5 – 318.2 culants (Ecotan and Tanfloc) with microalgae are used to treat
Spirulina sp. 12 220 413 urban wastewater in Spain done by Gutiérrez et al. The result
Thermosynechococcus 20 90 170 shows that more than 90% biomass recovery in 10 min for Ecotan
elongatus
and 20 min for Tanfloc. Hence, adding flocculants will shorten the
a
CO2 fixation rate¼ 1.833 $ biomass productivity which is derived from Chisti microalgae biomass sedimentation, and 10–50 g of Ecotan and
[17]. Tanfloc will not inhibit the subsequent anaerobic digestion

Table 10
Microalgae species used in wastewater treatment [16,31,83].

Species Source of waterwaste Total N removal Total P removal Carbon removal Retention time

(ChlorellaþNitzchia) Domestic wastewater 92% 74% 97% BOD and 87% COD 10 h
Chlorella pyrenoidosa Domestic wastewater 94% 80% – 13 days
Chlorella pyrenoidosa Domestic and industrial wastewater 60–70% 50–60% 80–88% BOD and 70–82% COD 15 days
Cyanobacteria Domestic effluent and swine wastewater 95% 625 – 1 day
Chlorella sp. Dairy farm effluent wastewater 75–83% 62–75% 27.4–38.4% COD 7–12 days
S.obliquus artificial wastewater 21a–53b% 45b–73a%
S.obliquus urban wastewater (indoor cultivation) 100% 98% – 183 h

BOD, biological oxygen demand.

COD, chemical oxygen demand.
a
Cultivating during summer season.
b
Cultivating during winter season.
194 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197

Fig. 14. Combination of wastewater treatment, microalgae culturing and HRAP and process flow [76,83,84].

Table 11
Advantages and disadvantages of culturing microalgae in wastewater treatment [16,83,85,86].

Advantages Disadvantages

Sufficient amount and ration of N and P in wastewater (total N:20–85 mg/L,total P:4–15 mg/L) and a More advanced technologies are needed to remove microalgae
substantial amount of C (TOC16:80–290 mg/L) from effluent (High cost)
Microalgae benefits wastewater treatment (toxic metals and nutrients removed) Flocculants can have a negative influence on the biomass to bio-
fuels process
Maximize CO2 capture Suspended solids limits are not met
Process has a lower energy demand Larger area needed than conventional wastewater treatment
Reduces pond area for microalgae culturing Not all process steps are commercially available
Lower capital and operating costs than conventional wastewater treatment Higher concentration of secondary metabolites means slower
growth of microalgae
Enhance methane gas production C/N ratio less than 20 will reduce the yield of methane gas

therefore the yield of methane gases remain the same (162 ml for
Ecotan 166 ml CH4/g VS) as without the use of flocculants [89].
Nevertheless, there are disadvantages of cultivating microalgae
in wastewater. Harvesting of microalgae in wastewater is difficult
and lack of carbons in wastewater reduce its efficiency of nitrogen
absorption by microalgae. Table 11 shows the advantages and
disadvantages of cultivating microalgae in wastewater [16,83,
85,86].
5.3. Microalgae biofuels: challenges and future prospects
Microalgae can produce a wide range of biofuels (diesel, gaso-
line, bioethanol and etc.) for transportation or can be directly
combusted for electricity generation. Microalgae crude oil has high
potential to replace petroleum since it has 80% of average energy
content of petroleum [79].
Growing microalgae for biofuels needed CO2 as carbon source.
At least 1.83 t of CO2 can be assimilated to produce 1 t of biomass.
However, for large-scale microalgae biomass production, nearly
50% cost is from CO2 purchased cost. Hence, cultivating microalgae
for fuels production is not feasible at the current stage [17,79]. In
microalgae/coal co-firing for electricity generation, CO2 production
from coal for microalgae utilization is still limited for commercial
scale [72]. Absorbing CO2 from atmosphere by microalgae could
not rapidly accumulate large quantity of microalgae biomass due
to diluted CO2 concentration [30]. Accelerating CO2 and seques-
tration in microalgae cultivation is possible by genetically mod-
ifying microalgae to evolve high activity carbonic anhydrase [90].
The limited light penetration in photo-bioreactors is one of the
reason resulted in difficulty to grow high density of microalgae at
commercial scale. Phagotrophic microalgae (e.g. Ochromonas
danica) are not limited by light source as their growth are
depending on ingesting bacteria [91]. O. danica can be effectively
used in wastewater system as cultivating of these microalgae does
not limited by light sources and suitable for high organic con-
centration medium. However there is not much research carried
out on phagotrophic microalgae but it might be sustainable in the
near future.
On the other hand, heterotrophic cultivation of microalgae is
only limited to a few species (e.g. H. pluvialis). However, this
 J. Milano et al. / Renewable and Sustainable Energy Reviews 58 (2016) 180–197
cultivation method always resulted in high biomass production
and high lipid content with simple bioreactor design (reduce cost
of production) [92]. Cultivation of heterotrophic microalgae will
eliminate the requirement of solar radiation and only required
certain concentration of organic compound as nutrients sources.
Nevertheless, heterotrophic cultivation will emit CO2 as by-
product and heavy contamination by other microorganisms [93].
Therefore, heterotrophic cultivation is restrictive in large scale
production even though this method resulted in large biomass and
high lipid content.
Energy ratio of biodiesel is a ratio of energy contained in
microalgae oil to input energy from fossil fuels. In cultivating and
producing microalgae oil, fossil fuels are usually used in pumping
the culture broth, filtration of water and other processes [79].
According to Khoo et al. [48], many estimations and assumptions
in literature are unrealistic. The oil content assumed in microalgae
is about 40% weights that will cost about $0.25 per kg compared to
petrol price which is $0.71 per kg. But the actual cost to produce
microalgae biomass is at least 10 times of this price [79].
Microalgae derived from biofuels have great potential to
replace fossil fuel and benefiting the environment. However, in
order to achieve high efficiency of microalgae biomass production,
continuous supply of solar energy and large amount of nutrient
are mandatory. In order to make the cultivation sustainable, the
nutrient can be supplied by the wastewater from food waste and
agricultural waste. However, pre-treatment of wastewater should
be performed prior to microalgae cultivation in order to reduce the
contamination. In addition, solar energy is only available at day
time, in which the growth of microalgae is retarded at night time
and required longer cultivation time to obtain optimum biomass
production. Integrated between phototropic and phagotrophic
microalgae with bacteria system can be performed to make
microalgae biofuels become more sustainable and cost competi-
tive with other biofuels. Moreover, harvesting of microalgae is also
a challenging task. This problem can be overcome by using natural
flocculants which successfully used by current wastewater system.
Further investigation to synthesize the natural flocculants should
be performed in order to reduce the time taken and cost for
microalgae harvesting system. Extraction of lipids from microalgae
depends on the current advance technologies to increase the lipid
extraction efficiency for biodiesel production. Simultaneous
extraction and reaction by using enzyme is believed to be alter-
native route to improve the overall biodiesel yield production. At
present condition, high cost of production and low energy ratio as
well as less integrating between technologies caused commercia-
lization of microalgae biofuels to be still in infancy stage [90].
5.4. Conclusion
Renewable energy technologies continue to expand and
received a lot of attention, particularly biofuels production from
microalgae biomass. Biofuels from microalgae biomass have the
potential to replace fossil fuel for power generation while miti-
gating CO2 emission from atmosphere. It is proven that chemical
compositions of microalgae fuels are similar to petrol and diesel
thus can be used to replace fossil fuels for transportation. In
addition, microalgae have high growth rates and lipid content
which make them a feasible choice for CO2 fixation and biodiesel
production.
Although microalgae have high potential to produce biofuels
and replace fossil in power generation, challenges to commercia-
lize the production at large scale need to be addressed. The chal-
lenges are such as: (a) the demand for biofuels is not high in
viewing low market price of fossil fuel; (b) the cost for the biofuels
production from microalgae is high; and (c) the energy ratio of
above unity needs to be achieved. Low cost with high efficient and
195
low contamination harvesting techniques are another obstacles
faced by the current stages of microalgae cultivation. This problem
can be solved by introducing natural flocculants, which is biode-
gradable with lower cost than mineral floccurants. Extracting the
oil from the dried microalgae is another challenge; even though
the microalgae are recognized of high lipid contents, but oil
extraction is not as anticipated.
Thus, biofuels production from microalgae biomass is con-
sidered expensive and the returns of investment are quiet slow
and low. However, with the improvement of advanced technolo-
gies and possible government incentives it is hoped that micro-
algae biofuels will soon become economic feasible and comparable
to fossil fuels in term of cost of production.
Acknowledgments
The authors would like to acknowledge the Ministry of Edu-
cation of Malaysia and University of Malaya, Malaysia for the
financial support under FRGS (FP009-2014A), HIR (D000015-
160001), UMRG (RP022A-13AET), PPP (PG017-2015A) and SATU:
(RU021A-2015).
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