Renewable and Sustainable Energy Reviews 64 (2016) 260270

Contents lists available at ScienceDirect

Renewable and Sustainable Energy Reviews

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

Botryococcus braunii for biodiesel production

Marija B. Tasi a,n,1, Luisa Fernanda Rios Pinto b, Bruno Colling Klein b,c, Vlada B. Veljkovi a,
Rubens Maciel Filho b
a
Faculty of Technology, University of Ni, Bulevar oslobodjenja 124, 16000 Leskovac, Serbia
b
State University of Campinas, UNICAMP, Laboratory of Optimization, Design and Advanced Control, Department of Chemical Process, School of Chemical
Engineering, Av. Albert Einstein, 500 CEP 13083-970, Campinas, Sao Paulo, Brazil
c
CNPEM/CTBE, Brazilian Bioethanol Science and Technology Laboratory, Division of Integrated Assessment of Bioreneries, Rua Giuseppe Mximo Scolfaro,
10000, Campinas, Sao Paulo, Brazil

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

Article history: This paper presents a review on microalga Botryococcus braunii (B. braunii) with an emphasis on lipid
Received 22 July 2015 production for possible industrial commercialization in biodiesel production. Summarizing the nature of
Received in revised form this alga is fundamental to establish the detail environment conditions for optimal biomass growth and
19 April 2016
the production of lipids, as well as upstream processing technologies for biodiesel production. This study
Accepted 15 June 2016
focuses the attention on identifying possible avenues for further investigation in the design of cultivation
systems and the integration of process technologies at industrial scales. The advantages and dis-
Keywords: advantages of biodiesel production processes based on these algal lipids are also analyzed. A particularly
Botryococcus braunii promising process involving B. braunii combines ue gas and wastewaters to produce lipids, converting
Lipids
them to biodiesel by supercritical (SC) direct trans/esterication (esterication followed by transester-
Biodiesel
ication). We conclude that such processes using B. braunii have considerable potential for the pro-
Cultivation
Harvesting duction of cleaner and more efcient biodiesel than current methods, reducing the environmental im-
Trans/esterication pact of fossil fuel use.
& 2016 Elsevier Ltd. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261
2. B. braunii as a biodiesel feedstock . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261
3. Toxic blooms and competition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 261
4. Growth environment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 262
5. Biodiesel technologies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264
5.1. Upstream pretreatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264
5.1.1. Cultivation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 264
5.1.2. Harvesting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265
5.1.3. Dewatering of harvested biomass . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265
5.1.4. Lipid extraction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 265
5.2. Trans/esterication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 266
5.3. Downstream purication. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 266
6. The quality of biodiesel obtained from B. braunii's lipids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 266

n
Corresponding author.
E-mail addresses: marijat14@yahoo.com, mtasic@tf.ni.ac.rs (M.B. Tasi).
1
Present address: State University of Campinas, UNICAMP, Department of Process Development and Products, Av. Albert Einstein, 500 CEP 13083-970, Campinas, Sao
Paulo, Brazil.

http://dx.doi.org/10.1016/j.rser.2016.06.009
1364-0321/& 2016 Elsevier Ltd. All rights reserved.
 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270 261

7. Economic assessment and life cycle analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 266

8. Conclusion and scope for future work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 267

1. Introduction braunii biodiesel production to identify technologies with im-

proved performance indicators. First of all, B. braunii is considered
The high carbon dioxide (CO2) concentration in the atmo- as a biodiesel feedstock referring to its nature, cultivation (in-
sphere, mainly the result of fossil fuel combustion, is thought to be cluding contamination problems and nutritional requirements),
one of the primary causes of global warming [1]. Biodiesel, being a harvesting, lipid extraction, and treatments to obtain FAMEs. Then,
renewable fuel, presents a promising alternative solution over biodiesel production processes based on these algal lipids are
fossil diesel. Biodiesel is typically generated by chemical reactions analyzed, highlighting both their advantages and disadvantages. It
(commonly known as trans/esterication) between lipids (e.g., is worth to emphasize that while relevant and readable reviews
triacylglycerols, abbreviated TAGs), free fatty acids (FFAs), and al- have been published on the biodiesel production from various
cohol (commonly methanol or ethanol), which generate fatty acid algal species [1114], none has included B. braunii.
alkyl esters. A variety of lipid-rich feedstocks can be used to pro-
duce biodiesel. These include: naturally derived crops; rened,
used or waste vegetable oils; animal fats; and algae lipids or 2. B. braunii as a biodiesel feedstock
sewage sludge. Algae are a particularly promising lipid source due
to their high lipid content and possibility to x atmospheric CO2 B. braunii is a green, pyriform shaped planktonic microalga,
through photosynthetic capture. They can also be cultivated on with colonies growing in the cluster and that can be found in
non-agricultural land or in aquatic environments, using waste- temperate or tropical oligotrophic lakes and estuaries, widespread
water as a nutrient source and without competing with food in freshwater and brackish lakes, reservoirs, ponds, or even
production or impacting land use. ephemeral lakes. B. braunii strains can be found in all climate
Algal biodiesel industrial production is still far from being zones except Antarctica [8]. Cell size (length
width) is in the
commercially viable, but it appears to be potentially suitable [2,3]. range of (89
5) mm to (13
79) mm [15]. Among various micro
Current problems include: (i) the considerable usage of non-agri- and macro algae species, B. braunii has been identied as the most
cultural land, fertilizers and water; (ii) high energy demands; (iii) promising for biofuel production, because of its excellent lipid
the relatively high up-front capital costs and the high price of production capability (content up to 65%, see Table 1). Although
biodiesel; and (iv) the poor transportation, storage, combustion, this microalga is known for high amounts of hydrocarbons and
and drivability performance of biodiesel. Several studies have ether lipids [16,17], its high content of saturated and mono-
pointed out that fatty acid methyl esters (FAME) present a variety unsaturated FAs as well as TAGs [18,19] makes it suitable for bio-
of problems when applied to automobile fuels [4]. However, the diesel production. The percentages of saturated, monounsaturated
efciency and quality of biodiesel production are heavily inu- and polyunsaturated FAs in B. braunii dry biomass revolve around
enced by the lipid extraction and lipid conversion methods used 9.85%, 79.61% and 10.54%, respectively [10]. Most of the FAs are
[57], and so the choice of feedstock is vital. The development of synthesized during the steady-state phase of algae growth, with
energy efcient and protable technologies through process in- the highest content of palmitic and oleic acids located in-
tegration is, therefore, a high priority for algal biodiesel tracellularly, but also in small amounts in the extracellular
production. medium.
The lipid content, the fatty acids (FAs) composition of TAGs and Lipid secretion in the extracellular medium is an unusual
quality (for example, iodine value (IV), cetane number (CN), FFAs phenomenon of B. braunii because all known species of microalgae
content) are the most important factors for selecting the right have cytoplasmic lipids. For example, the FAs lipids biosynthesis
species of microalgae. Polar and neutral lipids are preferred for the by B. braunii begins in the plastid, elongates within the en-
biodiesel production. Phaeodactylum tricornutum, Chlorella vul- doplasmic reticulum, transfers to the plasma membrane of the
garis, Chlorella protothecoides, B. braunii, and Nannochloropsis sali- endoplasmic reticulum or via Golgi bodies and then secretes to the
na are identied to produce such lipids [8]. Among all mentioned cell surface [46]. Lipid droplets are seen on the surface of the cell
species, B. braunii emerges as a promising lipid source because of apex [46]. The lipid yield and composition vary with the strain,
its widespread occurrence in aquatic environments and high lipid growth conditions and cell aging [4749].
content. According to an Aquatic Species Program (ASP) report [9],
B. braunii would not function well as a feedstock for the lipid-
based fuel production due to its slow growth (doubling time of 3. Toxic blooms and competition
72 h). The ASP also found this alga lipid to be less suitable for
transesterication (i.e. creating biodiesel), having most of its lipids The colonial B. braunii is a bloom-forming alga. The cause of the
as C29 to C34 aliphatic hydrocarbons, and less abundance of TAGs blooms and their subsequent damage to the populations of other
and C18 FAs. The FA proling of B. braunii (IBL-C117) is not ap- organisms has been widely studied [50,51]. Blooms are char-
propriate for producing a pure American Society of Testing and acterized with a high lipid content, about 40% of the dry biomass,
Materials (ASTM) nor the Brazilian Agency for Fuels and Biofuels compared with 18% for the culture collection strain [18]. Further-
(Agncia Nacional do Petrleo, Gs Natural e Biocombustveis, more, the lipid composition of bloom strains is represented mostly
ANP) grade biodiesel [10]. However, newer research shows that by saturated and monoenoic FAs. The dominant FAs are palmitic,
the doubling time could be reduced to 48 h and that both algal oleic, and linolenic acids. These FAs allow B. braunii to dominate in
lipid composition and FA proling are strongly affected by its en- natural environments due to their toxic effects on sh and zoo-
vironment growth conditions. plankton. Nonetheless, B. braunii's blooming nature is advanta-
The aim of this paper is to obtain a comprehensive picture of B. geous for biodiesel production due to quantity and quality of the
262 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270

Table 1
B. braunii strains of importance for biodiesel production.

Culture collection Strain Biomass Lipid contenta References

UTEX, USA LB-572 (96.47 6.8) g/Lb (26.80 72.05)% [20]

(0.0150.02) g/(L day)c 15.6%c [21]
0.304 g/(L day) 64.96% [22]
LB-572, A race 1.5 g/L 28% [23]
1.8 g/L 24%d [24]
UTEX 572 0.38 g/L 63% [25]
1 g/L 34.6% [26]
Austin 0.035 g/(L day) (46.85 75.20)% [27]
(0.026.55 7 7.66) (g/L d) 24% [28]
Institute of Hydrobiology, Chinese Academy of Sciences, PR China FACHB 357 5.5 g/(m2 d) 42% [29]
SAG culture collection, University of Gttingen, Germany SAG 807-1 6.45 g/(m2 d) 3.4412.44%e [30]
Gttingen 807/l Droop 0.032 g/(L day) (21.747 3.94)% [27]
Institute of Process Engineering, Chinese Academy of Sciences IPE 001 0.043.88 g/(L day) 46.39% [31]
Institute of Plant Physiology of the Russian Academy of Sciences Ktz No LB 807/1 Droop (0.16 70.01) g/(L d) 18.0% [18]
1950 H-252
1517%f [19]
0.7 g/L 22% [32]
Lake Shira (Khakasia, Siberia) A bloom of B. braunii, A g 40% [18,33,34]
race
Microalgal Culture Collection of the K.A. Timiryazev Institute of Ktz IPPAS H-252h,i 3.5 g/L 21% [35]
Plant Physiology RAS (IPPAS)
1.15 g/L (19.9 7 1.4)% [36]
Freshwater lake Kolleru, Andhra Pradesh, India Ktz. AP103j (1.7 70.051) (1.8 7 13) g/L 177 0.87 (19 7 0.98)% [37]
Ktz. AP104 (1.28 70.038) g/L (33079.9) mg/L
Ktz. AP105 (1.167 0.035) g/L (320 7 9.6) mg/L
Centre for Advanced Studies in Botany, University of Madras, Chen- AP102 1.9 g/L 48.40% [38]
nai, India
Natural Brazilian ecosystems LEM 14 1.88 g/L 36.14% [39]
Biogeochemistry and Ecology of Terrestrial Environments Depart- A race 0.46 g/(L day) 17.85% [40]
ment at the cole Nationale Superieure de Chimie de Paris
Lake of Coat at Herno, Brittany A race 60.6% [41]
UTEX, USA Austin A race 27.4 g/100 g [42]
Fresh water UTEX 572 (0.037 70.005) (g/L d) (13.50 7 3.78)% [43]
Lake in Trag province, Thailand TRG 5.16 g/L 37.5% [44]
Freshwater reservoir in Assam, India GUBIOTJTBB1 0.316 g/L (57.147 1.18)% [45]

a
Total lipids.
b
Immobilized system cultivation.
c
Outdoor production under uncontrolled conditions.
d
Intracellular lipids. The content of total extracellular lipids is not available.
e
Non-polar lipids.
f
Raceway pond cultivation.
g
Not available.
h
Due to hydrocarbon and fatty acid composition this strain is closer to Botryococcuss udeticus Lemmermann.
i
Also known as B. braunii Ktz No LB 807/1 Droop 1950 from Culture Collection of Algae and Protozoa, Dunstaffnage, Scotland.
j
Open raceway pond conditions.

synthesized lipids. 4. Growth environment

Because of the relatively slow growth of B. braunii, trials to
cultivate B. braunii in non-axenic systems often fail due to biomass
contamination from other green algal and cyanobacterial species.
B. braunii's competitors include Chlorella vulgaris, Chlamydomonas
reinhardtii, Scenedesmus dimorphus, Pseudokirchneriella sub-
capitata, and Merismopedia tenuissima [52]. Contamination among
algal species is a consequence of competition among various or-
ganisms for the critical nutrient in the medium. Furthermore, this
contest is mainly inuenced by the biomass ratio of competing
species. For example, the low or medium ltrate density of two
common algae species in natural waters - Chlorella vulgaris and
Chlamydomonas reinhardtii - stimulates B. braunii growth, while
high cell ltrate density cultures signicantly inhibit it [53]. The
presence of bacteria is also associated with the mass culture of B.
braunii. For example, the gram-negative Rhizobium sp. [54] forms a
biolm (microalgal matrix) and not only signicantly stimulates B.
braunii growth but also produces the cell-to-cell signal molecules
known as acyl-homoserine lactones. The formation of B. braunii's
bloom is also one of the outcomes for contamination caused by
other microorganisms.
The major target in industrial B. braunii biodiesel production is
increasing biomass production while improving the quantity and
quality of the lipid product in a sustainable way. Carbon, nitrogen,
phosphorus, sulfur, and iron are essential nutrients both for bio-
mass productivity and for the quantity and quality of the synthe-
sized lipids. To reach this target, many synthetic media have been
investigated [20,23,24,28,29,39,5560], but only the modied
Chu13 medium [29,30,61] has been shown to be optimal. How-
ever, commercial biodiesel production using this synthetic med-
ium is mostly discouraged, as the high consumption rates of fossil-
based fertilizers as the source of inorganic nutrients make it costly
and unsustainable. It means that the modied Chu13 medium is of
importance only for B. braunii inoculum preparation at commercial
scales.
Recent research has been directed towards the cultivation of B.
braunii in natural media. For example, since the natural habitat of
B. braunii is seawater, recently published papers suggest the pos-
sible exploitation of NaCl-rich seawater for the outdoor commer-
cial cultivation of B. braunii [62,63]. However, not all B. braunii
strains are halophilic. The biomass yield and the lipid content of B.
 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
braunii (LB 572) increase under all tested NaCl concentrations
(0.0170.085 M), reaching the highest content of palmitic and oleic
acid at 0.085 M [23]. Higher concentrations of NaCI (0.50.75 M)
are either toxic to the Austin and Gottingen strains, respectively
[27] or decrease the biomass concentration and the lipid yield of B.
braunii KMITL 2 [64]. Zhila et al. [36] show that haloadaptation is a
specicity of the strain growth phase. For example, the presence of
salt (0.3 M and 0.7 M) in the medium inhibits the growth of B.
braunii Ktz (IPPAS H-252), enriching TAGs and oleic acid content
for the rst three days, and increasing biomass and polyenoic FAs
content in later growth phases [36]. The cultivation of B. braunii in
seawater shows promising results in reducing the CO2 content
since xed atmospheric CO2 [38,44], and industrial exhaust gasses
[28] are often used as the carbon sources under photoautotrophic
conditions. However, the CO2 fraction in the air of about 0.03% [44]
is not sufcient. Hence, an extra amount up to 2% (v/v) is needed
to obtain increased biomass (1.31.8 times) and palmitic and oleic
acid (2.53 times) concentrations in B. braunii (LB 572) strain cells
[65]. Of greater interest from the environmental and the biodiesel
production point of view is the use of industrial ue gas as a CO2
source, since B. braunii tolerates [66] the sultes and bisultes that
are usually present in the ue gas. It is corroborated by the fact
that at the ue gas rate of 0.3 v/v/m, the biomass and lipid (with
an oleic acid fraction of 55%) productivity for B. braunii increases
2.9-fold and 3.7-fold, respectively, compared to cultures cultivated
with ambient air containing 10% CO2 [28]. However, using ue gas
for the B. braunii cultivation in seawater impacts the marine eco-
system, coastal area shing, boating and ship trafc.
Wastewaters are promising B. braunii cultivation media due to
their enhanced nitrate and mineral contents. As can be seen in
Table 2, wastewaters are rich in various nitrogen sources, such as
nitrate, ammonium, and nitrite. An earlier investigation showed
that B. braunii rst consumes the nitrogen sources with the
highest concentration in the medium [44]. However, when their
concentrations are equivalent, ammonium and nitrate uptakes are
prioritized [67]. Furthermore, low nitrate levels in the nutrient
medium (below 0.04 mM) lead to lower B. braunii biomass growth
rates, higher lipid content and lower lipid productivity of
0.090 day  1, 63% and 0.009 (g/(L day)), respectively [25]. This
lower biomass production has a negative effect at industrial levels,
where lipid productivity is more important than lipid content [21].
High B. braunii (UTEX 572) biomass growth rates and lipid pro-
ductivities of 0.159 day  1 and 0.019 g/(L day), respectively can be
achieved at a moderate nitrate concentration of 0.37 mM [25].
However, a B. braunii (KMITL 2) strain showed a lower lipid pro-
ductivity rate of 0.004 g/(L day) [64] with an almost equal nitrate
Table 2
Common chemical composition and nutrient concentrations of various wastewaters (mg/L).
Ion Efuent from a secondary domestic Efuent from a secondary urban
wastewater treatment system wastewater treatment system
2
Na 28.98
NH4 12.76 15.00
K 7.99
Mg2 2.63
Zn2 0.71 o0.05
NO3  0.2 0.90c
PO43  2.00 11.50
Cl  1.70
SO42  4.21
References [39] [40]
a
Also contains small elemental phosphor and sulphur in both untreated and treated wastewater samples.
b
Also contains NO2  of 0.04 mg/L.
c
In total nitrogen content of 11.9 mg/L.
263
concentration (0.42 mM). It was explained by the fact that the B.
braunii (KMITL 2) strain was not pre-cultured before medium in-
oculation, which would have helped the algal cells to reproduce
further.
Phosphorus and sulfur concentrations also have major impacts
on both the growth and lipid productivity of B. braunii (LB 572), as
demonstrated by statistical optimization of culture media [56].
Phosphorus - usually in the form of phosphate - at concentrations
up to 444 mg/L increased the biomass concentration and reduced
the lipid content 7.3 fold and 1.7 fold, respectively [64]. Lipid
productivity also increased 1.2 fold by enhancing the phosphate
concentration from 40 mg/L to 83 mg/L [56]. However, it should be
pointed out that phosphate was not a limiting factor to biomass
growth [17] due to the B. braunii's ability to accumulate large
amounts of surplus phosphorus [68]. The source and concentra-
tion of sulfur also inuence the growth of B. braunii. Concentra-
tions of bisulte, sulte and sulfate up to 1 mmol/L, 3.2 mmol/L
and 4 3.2 mmol/L, respectively showed no toxic effects on the
growth of B. braunii [32]. However, these experimental data did
not indicate the inuence of sulfur source and concentration on
the lipid content or productivity. Although not reported in Table 2,
iron usually as FeCl3 at concentrations up to 0.74 mM - decreases
biomass productivity under nitrogen-decient conditions [57].
Additionally, it increases the lipid productivity (with highest
fractions of palmitic and oleic acids) of other B. braunii strains
(TRG, KB, SK and PSU). Other elements such as potassium, mag-
nesium and zinc are known to have an impact on algae cultivation
but have not been reported to be signicant for B. braunii biomass
and lipid productivities. By comparing these nutrient values with
the reported ones (Table 2), we can predict the optimal medium
for both the biomass and lipid (rich in palmitic and oleic acid
content) productivities. For example, efuent from a secondary
domestic and untreated seafood wastewater treatment systems
are optimal for B. braunii (LEM14) and B. braunii (UTEX 572), re-
spectively. Wastewaters can also be sources of organic carbon. The
cultivation of B. braunii in a molasses wastewater produced 1.3 and
1.2 times more biomass and lipids compared to photoautotrophic
growth [44]. From the environmental point of view, the effect can
be enhanced by adding both organic and inorganic carbon sources
when cultivating B. braunii, which can solve the wastewater and
industrial exhaust gasses problems. For instance, the CO2 supply in
a BG-11 medium enriched with glucose provides higher biomass
and lipid yields [69].
The optimal biomass and lipid productivities in synthetic and
natural media can be obtained by controlling several para-
meters: temperature, pH, light and mixing. Excepting two strains
Mixa of carpet industry wastewaters Secondary seafood processing
and sewage plant wastewaterb
Untreated Treated
112.1133.4 117.8151.5
17.5825.85 0.573.61 4.6
7.7510.32 9.7410.17
13.8220.78 11.2813.21
0.030.13 0.170.18
0.2128.13 1.393.91 24.1
20.3135.10 17.5921.95
86.96142-80 128.26182.10
[43] [44]
264 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
Table 3
Comparison between typical microalgae cultivation systems.
Open systems Closed systems
Advantages 1. Relatively cheap 1.
2. Easy to clean 2.
3. Utilizes non-agricultural land 3.
4. Low energy inputs 4.
5. Easy maintenance 5.
6. Easy to scale-up 6.
7.
8.
Disadvantages 1. Low productivity 1.
2. Environmental factors are beyond
control 2.
3. Large area of land required 3.
4. Limited to few strains of algae 4.
5. Signicant evaporative losses 5.
6. Poor Stirring, light and CO2 utilization
7. Cultures are easily contaminated 6.
8. Limited recovery of biomass
(BOD-NG17 and BOD-GJ2) that show tolerance to extreme tem-
peratures of 40 C and  20 C [32], most B. braunii strains have an
optimal growth temperature of 25 C and show satisfying re-
sistance to temperature changes. This temperature tolerance can
be substantial for the B. braunii outdoor mass cultivation because
problems like overheating and/or poor thermal (temperature)
control can be avoided. Furthermore, the temperature change
correlates strongly with CO2 solubility and indirectly with pH
maintenance. Higher temperatures decrease CO2 solubility and
increase pH, which favors the blooming of B. braunii [50,51]. On
the contrary, the reduction of pH results in a low biomass pro-
ductivity [44,65]. A wide pH range of 6.08.5 was tolerated only by
two B. braunii strains (LB-572 and SAG 30.81) [70] while the
adaptation period of B. braunii (UTEX 572) was longer [28]. Ge
et al. [71] reported that the B. braunii (765) strain performed os-
moregulation by the synthesis/dissimilation of intracellular gly-
cerol, which maintained constant intracellular pH over a wide
range of the external pH. However, there is no data conrming
that lipid production by this algal strain is affected by the culture
pH. For outdoor algae cultivation, besides temperature and pH,
solar radiation and light path have major impacts. The average
recorded solar radiation during the outdoor cultivation of B.
braunii is about 5 kW h/(m2 day) [21,37]. At constant solar radia-
tion, higher biomass culture concentrations can be achieved if the
light path is shorter [21]. On the other hand, if articial light is
used with a 16:8 light:dark cycle, uorescent lamps are most ef-
fective for increasing B. braunii (KMITL 2) lipid contents and yields
[64]. By contrast, blue-green LEDs lower costs for B. braunii (NIES-
836) cultivation [72]. However, improved pH, temperature dis-
tribution and light intensity conditions are ineffective without
appropriate mixing. For B. braunii strains, mixing is most fre-
quently provided by CO2 aeration [69,71,73,74] or mechanically
using a 15100 rpm velocity paddle wheel [24,38]. Compared to
manually tested cultivations, the rapid (80100 rpm) paddle wheel
mixing provided higher lipid content of 48.4% [38] and lower oleic
acid content (19.2%). It leads to the conclusion that the highest
biomass and lipid productivities were obtained under lower mix-
ing intensity.
5. Biodiesel technologies
Algae biodiesel technology includes three phases: upstream
pretreatment, trans/esterication and downstream purication.
The upstream pretreatment consists of the following steps: culti-
vation in open and closed systems, harvesting, dewatering of
Hybrid systems
High biomass productivities 1. Minimal contamination due to controlled inoculum growth
Suitable for outdoor cultures in the rst stage
Good stirring 2. Sufcient seed supplies keep the preferred algal specie
Avoid water evaporation (s) dominant
Well controlled culture conditions
Permits single strain culture
Increased the amount of light received
Lower level of contamination
More expensive to construct and 1. Large-scale applications have been limited by the cost of the
operate rst stage
High energy inputs
Requiring cooling
Not easy scalable
Labor intensive maintenance of bag
systems
Not eliminates contamination
harvested biomass, and lipid extraction. The downstream pur-
ication steps are: possible catalyst separation, alcohol recovery,
nal product purication, and wastewater treatment. While the
downstream purication steps are usually common for various
technologies, the upstream pretreatment phase is of crucial im-
portance, and hence, it is further discussed.
5.1. Upstream pretreatment
5.1.1. Cultivation
B. braunii can be grown in aquatic environments or in non-
agricultural land cultivation systems located near power plants
and/or wastewater treatment plants to access CO2 from ue gas
and/or nutrient supplies, respectively. The goal of B. braunii culti-
vation is to mimic the natural environmental conditions which
favor algal growth and lipid productivity. Generally, according to
their design characteristics, B. braunii cultivation systems can be
classied as open or closed.
Open systems are outdoor facilities such as ponds (unstirred,
inclined, with a paddle wheel, and circulating ponds), lagoons,
deep channels, and shallow circulating units. Closed systems, also
known as photobioreactors, include vessels, vertical-columns, os-
cillatory ow reactors, airlift reactors, xed or oating, helical
(coils) or annular tubes, plates (panels or layers), and bags - all
with walls made of transparent materials. Photobioreactors can
also be located outdoors, under sunlight or indoors, under articial
irradiation facilities. In closed design systems, algae can also be
immobilized. Currently, there are six types of immobilization
methods: covalent coupling, afnity immobilization, adsorption,
connement in the liquid-liquid emulsion, capture behind semi-
permeable membrane and entrapment. Five types of bioreactors
are used: uidized-bed, packed-bed, parallel-plate, air-lift, and
hollow-ber.
Hybrid systems, which are a combination of open and closed
systems, are also reported in the literature [7577], but not for B.
braunii cultivation. Advantages and disadvantages of each culti-
vation system are summarized in Table 3.
5.1.1.1. Open systems. Four strains of B. braunii have been cultivated
in open systems for biodiesel production. B. braunii (AP103), B.
braunii (LB 572) and two B. braunii strains isolated from Loktak and
Udaisagar lakes were cultivated in raceway pond [24], raceway and
unstirred circular ponds [37] and plastic trays [37], respectively.
Size capacities were: 0.710 m2 (2032000 L) and 1 m2 (287 L) for
the raceway and unstirred circular ponds, respectively. All these
experiments were photoautotrophic cultivations on synthetic
 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
(modied Chu 13) medium with atmospheric CO2 as an inorganic
carbon source, thus making commercial applicability limited.
However, the cultivation time of 18 days was enough to produce
approximately 1.85 g/L of B. braunii biomass, and mixing had the
main inuence on the lipid product quantity and quality. The
optimal mixing rate was 15 rpm [24], which induced 2024% of
lipid content with 34% of oleic acid. Mosquitos can severely affect
B. braunii outdoor biomass production if the nutrient medium is
not supplemented with 0.5 M NaCl [63].
Currently, open systems are not implemented on commercial
scales for the B. braunii cultivation intended for biodiesel pro-
duction, due to numerous unanswered questions. For example:
(i) Can we improve the low productivity of open systems by im-
plementing nutrient-rich wastewaters, using large inoculum or
frequent feeding/harvesting. (ii) Does the covering of open ponds
with permeable plastic covers or greenhouses provide better
control over water evaporation, atmospheric temperature, wind
speed, net solar radiation as well as transfer and supply of CO2?
(iii) What is the quality of water obtained after algal biomass de-
watering? Is it compatible with the optimal growth composition?
(iv) Is the Submersible aquatic algae cultivation technology [67]
adaptable for the B. braunii strains?
5.1.1.2. Closed systems. Photobioreactors implemented in B. braunii
production use both suspended and immobilized microalgae cells.
Suspended B. braunii (LB572) and B. braunii (IPE 001) cells are
cultivated in the panel [21] and plastic bag [31] reactors, respec-
tively. Also, the cultivation of suspended B. braunii (NIES 2199)
cells in a submerged membrane photobioreactor using efuent
off-gas (CO2) from an aeration tank and nutrients from the treated
sewage was modeled [78]. Novel immobilized cultivation systems
with thick concrete layers and wood support plates [36] or biolm
layers [29,30] are also used for cultivating B. braunii (LB 572), B.
braunii (FACHB 357) and B. braunii (SAG 807-1). Capacities of 400
1000 L and 200 L for the panel and plastic bag reactors, respec-
tively, have been used only for suspended cells while immobilized
cells are still at the laboratory scale. Like open systems, all of these
experiments were conducted using synthetic media with atmo-
spheric CO2 as an inorganic carbon source [31] and using fertilizers
[21]. Immobilized biolm cultivation gives much lower biomass
productivity compared to open pond systems, but the highest one
(5.5 g/(L day)) among closed systems. However, the maximum li-
pid productivity of 20.36 g/(L day) was obtained with a plastic bag
photobioreactor under outdoor ultrasonic-treated conditions. All
of these studies indicated that linoleic and oleic acids are major
lipid components.
Although the use of non-agricultural land for closed systems is
far less than for open systems, current photobioreactor technolo-
gies have not yet been implemented for cultivating B. braunii on
commercial scales. For example, outdoor plastic bag cultivation
faces problems with high operating costs, limited recovery of
biomass, and light availability. On the other hand, limitations of
immobilized biolm cultivation include the high density of im-
mobilized matrix reducing the light penetration, its high cost and
limited scalability beyond the laboratory. A very interesting closed
system design for cultivating green microalgae (Desmodesmus sp.)
using wastewaters is the Offshore Membrane Enclosures for
Growing Algae (OMEGA) system [79]; however, there has been no
report on its application for B. braunii cultivation.
5.1.2. Harvesting
Microalgae harvesting is a two-stage process involving: (i) bulk
harvesting, such as occulation, otation, or gravity sedimenta-
tion; and (ii) thickening, such as centrifugation, ltration, or ul-
trasonic aggregation [8082]. Harvesting is a challenging phase
due to the low densities and small sizes of algal cells, as well as the
265
high energy and total costs involved. The advantages and dis-
advantages of each harvesting technique are well summarized in a
recently published paper [83]. Since B. braunii grows in colonies,
its harvesting is less challenging than for no-colony microalgae
[84]. So far, laboratory scale occulation [26,85] and in situ mag-
netic [31] harvesting have been tested for B. braunii cells rich in
lipids. B. braunii (UTEX 572) cells had been harvested using various
occulation methods. The pH adjustment and addition of various
salts such as Al2(SO4)3, NH4Cl, KCl, CaCl2, MgSO4, and FeCl3 [26,85]
or bioocculants such as Pestan and that produced by the bac-
terium P. polymyxa AM49 [26,85] are just a few noteworthy. The
most efcient method for recovery (with the occulating activity
of 90.6%) was treatment with both FeCl3 and the bioocculant
produced by the bacterium P. polymyxa AM49. The same recovery
efciency of 90% was achieved with an in situ magnetic separation
system [31]. Although the former method has relatively low cost
and no cells damage, it has the following major challenges for
commercial applications: high energy inputs, the possibility of
bioocculants agents competing for medium nutrients, and the
inability to recycle water back into the culture system. On the
other hand, in situ magnetic harvesting suffers from high pro-
duction costs associated with the high cost of CO2 consumption
and capital investments. Therefore, different harvesting techni-
ques must be integrated to increase the efciency of microalgae
harvesting. For example, electro-occulation and dispersed air
otation were integrated for harvesting B. braunii [86] to improve
the biomass recovery signicantly. The recovery efciency of 98.9%
was reached within only 14 min. Future investigations should
determine the energy and cost demands of this method as well as
the possibility for its scaling-up.
5.1.3. Dewatering of harvested biomass
Some processes, such as in-situ transesterication or hydro-
thermal liquefaction (HTL), need no or only partial dewatering of
harvested biomass. In all other cases, extensive biomass dewater-
ing is required due to the possible inhibition of subsequent pro-
cesses such as lipid extraction and trans/esterication. Also, de-
watering temperature strongly affects both lipid composition and
yield. Commonly used methods to dry the wet biomass paste are:
solar, low-pressure shelf, spray, drum, uidized-bed, freeze, and
Refractance Window drying [80]. However, dryers are high
energy-consuming devices and, therefore, this type of dewatering
of harvested biomass can seriously jeopardize the commercial
viability of microalgae biodiesel production. Little information
about the development of efcient dryers for the B. braunii bio-
mass is available in the literature.
5.1.4. Lipid extraction
For feasible extraction of lipids, a good starting point is to
measure total lipids from B. braunii using Nile Red (9-diethyla-
mino-5H-benzo[]phenoxazine-5-one) staining [87]. After this
inexpensive and uncomplicated method, lipid recovery can be
carried out in two steps that include cell disruption and lipid ex-
traction. There are numerous disruption methods, which are di-
vided into four categories: mechanical, physical, chemical, and
enzymatic. Each category is widely explained and well summar-
ized elsewhere [88]. Since lipids of B. braunii are accumulated in
vacuoles, they can easily be released by slight cell disruption. At
the laboratory scale, B. braunii cells are mainly disrupted me-
chanically [60]. It seems that this disruption method does not af-
fect the extracted lipid yield since the same yield is obtained with
lysed and whole cells. The effect of this disruption method on the
lipid composition has not yet been investigated.
Lipid extraction methods, such as solvent extraction, SC uid
extraction, HTL, and milking in microalgal biodiesel production are
worthy of mention [60,8992]. The advantages and disadvantages
266 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
of different methods for lipid extraction have been widely studied
[93,94]. Amongst all, SC uid extraction is the most efcient for B.
braunii lipid extraction. Similar FAME yields per ash-free dry
weight (AFDW) of 70.85 mg/g AFDW and 60 mg/g AFDW [60] were
obtained from lipids extracted with hexane and SC CO2 extraction
methods, respectively. Hence, the latter was shown to be more
selective for the extraction of neutral lipids [60,95], which are
necessary for biodiesel production. Also, the harvested B. braunii
cells can be thermochemically liqueed, yielding an oil (64%),
aqueous, gaseous (CO2), and tar fractions [91]. However, in this
research, the oil was not characterized, so it is questionable if it
can be applied to biodiesel production. It is interesting to mention
the milking of B. braunii Ktz (CCAP 807/2) cells in both solvent-
based and the solvent-free (blotting) forms, as they are non-de-
structive external oil extraction methods which cause no cell da-
mage [90]. It means that after lipid extraction, the cells can be
returned to the cultivation medium to continue synthesizing li-
pids. However, this method is not signicant for biodiesel pro-
duction due to low TAG yields (o6.65 mg/g AFDW and
o0.08 mg/g AFDW for the 20 min heptane and the 3 h blotting
methods, respectively). However, since milking efciently extracts
hydrocarbons (495%) it is of enormous importance for obtaining
renewable, white and Hydro-Thermal-Upgraded (HTU) diesels.
Currently, SC uid extraction is only conducted at pilot scale
because of limitations caused by high energy and economic costs.
However, scale-up may not be a necessary step, since quantities
produced in short timeframes from cultivators are in kilogram-
scale. Lastly, it remains to be seen how efcient one-step dewa-
tering, cell disruption, and lipid extraction techniques, such as the
Origin Oil single step extraction method [92], will be for B. braunii
lipid extraction in biodiesel production.
5.2. Trans/esterication
After the extraction of lipids from microalgae, the trans/ester-
ication process is necessary to produce high-quality biodiesel.
Trans/esterication is the most studied biodiesel production phase
of the last decade. Comprehensive pathways of microalgae bio-
mass trans/esterication have already been investigated
[73,81,83,9698]. Methanol and ethanol are frequently used,
especially the former one, for producing biodiesel and glycerol.
The trans/esterication process may occur in the presence of a
base, acid, enzyme, or with no catalyst. The optimal reaction time
to achieve a high lipid conversion depends on the operating con-
ditions and the catalyst type and varies from one minute [99] to
two days [100]. Due to the incomplete lipid extraction from the B.
braunii biomass (high contents of residual biomass rich in lipids),
improvements to existing methods are directed towards the in-
tegrated concept of simultaneous extraction and trans/esterica-
tion, also known as direct trans/esterication or in-situ trans/es-
terication (ISTE) [89,101]. ISTE in batch and reux extraction re-
actors in semi-continuous laboratory scales are performed under
solvent/cosolvent/catalyst [89,101] conditions. The highest FAME
yield of 480% is obtained in a reux extraction reactor using the
homogenous H2SO4 catalyst and a low solvent/cosolvent ratio (3:1
methanol/chloroform). ISTE is a single-step process which reduces
both capital and energy costs. Moreover, it is most effective for
measuring total fatty acids in microalgae [89], which are essential
for biodiesel production. However, the high biomass water content
signicantly decreases the biodiesel yield, and the presence of
cosolvents/catalysts makes downstream processing more complex.
Although it is not employed in the case of the B. braunii, ISTE under
SC conditions [102] is a good alternative to classic ISTE. The SC ISTE
method is a one-step process for direct liquefaction and conver-
sion of wet algal biomass without using cosolvent/catalyst mix-
tures. However, the techno-economic feasibility of SC ISTE and its
possible improvements are yet to be studied.
5.3. Downstream purication
For commercial acceptance, biodiesel produced from micro-
algae needs to meet currently existing fossil fuel standards. It
means that biodiesel needs to be separated from the trans/ester-
ication reaction mixture and further puried to reach the stan-
dard quality. Technologies such as gravitational settling, micro-
wave irradiation, centrifugation, membrane ltration, wet/dry
washing, and membrane rening are often included in biodiesel
separation and purication [103106]. However, separation and
purication studies of B. braunii biodiesel have not been revealed
so far. Furthermore, the biorenery concept must use a zero-waste
process, so the utilization of by-product process streams is crucial
for plant facility design. If the process ows - for example, lipid
extracted microalgal biomass residues - can not be recycled into
the process, they should be converted into biofuels like bio-
methane, bioethanol, biohydrogen, biobutanol [107] or other va-
luable products like drugs, food, feed additives [108], and animal
feed [109]. The use of combustion, gasication, pyrolysis, lique-
faction, carbonization, and anaerobic digestion of the algal bio-
mass residue are widely cited in the literature. Nevertheless, the
lipid-extracted residue biomass of B. braunii (BOT-22) has been
investigated only as a feedstock for steam pyrolysis producing a
hydrogen-rich solid fuel which is comparable to fossil fuel [110].
6. The quality of biodiesel obtained from B. braunii's lipids
Although this alga is mainly studied for hydrocarbon produc-
tion, recently published papers conrm its application for biodie-
sel production, too. A signicant number of biodiesel physico-
chemical properties were observed to be similar to that of petro-
leum diesel fuel. For instance, B. braunii (UTEX and IBL-C117)
showed better matching with all three International standards
(EN14214, ASTM D6751-10, RANP/2008) values regarding CN, cold
lter plugging point (CFPP), and (IV) [111]. Even better fuel prop-
erties results were found in B. braunii (KMITL) strain due to lower
IV and degree of unsaturation (DU) values which are signicant for
oxidative stability of biodiesel [112]. However, all three B. braunii
strains were cultivated in synthetic media which inuence the FAs
proling and consequently the quality of biodiesel. Hence, a major
boost in the technology of B. braunii biodiesel synthesis will be
producing high-quality fuel from natural media which will meet
International standard directives.
7. Economic assessment and life cycle analysis
The biggest obstacle for the commercial production of biodiesel
from algal biomass is its high price (US$ 2.76/kg), which is nearly
triple that of fossil diesel (US$ 0.95/kg) [113]. This price is due to
complex, hardly scalable and costly processing steps, such as cul-
tivation, harvesting, dewatering of biomass, and lipid extraction.
Table 4 compares the cost of various production phases using B.
braunii and other microalgal species.
This table indicates that the costs associated with open ponds
are the lowest. Furthermore, the crucial economic problem to be
solved is to develop low-cost harvesting and lipid extraction
methods. It is because harvesting, drying and extracting techni-
ques represent about half of the total production cost [97]. Har-
vesting alone amounts to 30% of the total cost of biodiesel pro-
duction from microalgae [14] and can be as high as about US$ 10/
kg [73]. To be competitive with fossil diesel, the cost of extracted
algal lipids should be:
 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270 267

Table 4 each nutrient on lipid composition (especially FAs and TAGs) is a

Costs of the production phases for B. braunii and some other algae. major disadvantage. Also, a systematic approach to optimizing the
medium nutrition composition for the selected producing strain is
Costs References
necessary for its commercial utilization. It means that much ex-
Cultivation B. braunii (IRE 001) US$ 20.76/kga [31] perimentation is required to determine the inuence of each nu-
Reports on other taxonomic US$ 1.98/L to US$ [114] trient type and concentration on the biomass and lipid pro-
groups of algae 5.50/Lb,c ductivities, as well as on the lipid composition of each growth
US$ 3.30/L to US$
8.80/Lb,c
phase for the specied strain of B. braunii. This information will
US$ 0.60/kg and US$ [115,116] help in concluding if a natural medium, such as wastewaters, is
1.54/kgc useful as the cultivation medium for B. braunii strains which will
US$ 0.47/kg and US synthesize lipids for high-quality biodiesel.
$ 7.32/kga
The design of cultivation systems has also been widely studied.
US$ 1.02/kgd [116]
The open pond is proven to be viable for industrial cultivation of B.
braunii. The problems associated with the open pond cultivation,
Harvesting B. braunii (IRE 001) US$ 2.07/kge [31]
such as contamination or unsustainable supply of fertilizer, can be
Reports on other taxonomic US$ 0.07fUS$ 0.22/ [117]
groups of algae Lg minimized. It is because B. braunii: (i) grows in colonies; (ii)
US$ 0.00011/Lh [86] blooms, suggesting allelopathic effects; and (iii) can be easily
grown in wastewaters rich in nutrients with a supply of CO2. Also,
Extraction Reports on other taxonomic US$ 0.02US$i 0.06/L [117] medium densities of Chlorella vulgaris and Chlamydomonas re-
groups of algae inhardtii stimulate the growth of B. braunii. However, the con-
US$ 1.87/L [118] siderable amount of land for open ponds, the low biomass yields
US$ 2.80/L [115] and the evaporation should be addressed in the future works.
US$ 3.98/L [118]
Closed loop systems do not have the problems of open-pond
a
Photobioreactors. systems, but they cannot easily be scaled, are not cheap to con-
b
Biodiesel price. struct and consume more energy. New methods for B. braunii
c
Open ponds. biomass harvesting and lipid extraction should further be in-
d
ermenters.
e vestigated due to their signicant shares of the total production
In situ magnetic harvesting.
f
Algae Venture Systems's harvesting technology. costs. Reducing the number of steps by introducing integrated
g
Centrifuge. phases, such as SC ISTE, is essential to provide easier, better and
h
Electroocculation. lower cost systems. Also, while laboratory experiments have
i
Algae to Energy Inc. patented system. mostly been reported for harvesting and trans/esterication, these
production phases are yet to be tested further at the pilot and
Calgal lipid = 8.48104 Cfossil diesel (1) industrial scales. The biodiesel must be thoroughly examined re-
garding its quality as a fuel, and no hazardous wastes should be
where, C algal lipid is the price of algal lipid (US$/L) and C fossil diesel is released during downstream processing. It means that more work
the price of crude fossil diesel (US$/L), assuming that algal lipids on the economic feasibility and LCA of biodiesel production from
have 80% of the caloric value of crude fossil diesel [115]. It means B. braunii should be done.
that a reasonable algal lipid price should be US$ 0.48/L. To the best
authors knowledge, the costs of B. braunii extracted lipids have
not been reported yet. Acknowledgments
Besides economic benets, the ecological footprint obtained by
life cycle analysis (LCA) is another advantage and conformation of This study was supported by Fundao de Amparo Pesquisa
the environmental sustainability of microalgae-based biodiesel. do Estado de So Paulo (Process No. 14/18979-6) and the Ministry
Among a few LCA reports considering algal species [119123], of Education, Science and Technological Development of the Re-
there is one dealing with a report on the LCA for B. braunii used in public of Serbia (Project III 45001). The authors are particularly
hydrocarbon production [124]. thankful to Dan S. Taranu, Ph.D. and Marija D. Stankovi, Ph.D. for
revision of the English and additional comments.

8. Conclusion and scope for future work

References
For industrial output and commercialization of high-quality
biodiesel from microalgae B. braunii, the amount and composition
of lipids are important, assuming no signicant change in their
composition during processing. Both the amount and composition
of lipids depend on the strain and cultivation conditions. In gen-
eral, B. braunii accumulates TAGs and saturated FAs initially under
stressful cultivation conditions while adjusting to the new en-
vironment. Later, in the exponential phase, hydrocarbons are the
main components, which are excreted in the extracellular space
favoring the cells survival. Finally, in the stationary phase, the cells
favor the synthesis of TAGs and saturated FAs, which are of im-
portance for biodiesel production.
Previous works on B. braunii has been focused on the opti-
mizing the growth environment and favoring rapid biomass
growth and higher lipid accumulation. Generally, from the bio-
diesel production point of view, the lack of data on the inuence of
[1] United Nations. Climate Change Information kit. Clim Chang Inf Kit Int En-
viron House, Geneva, Switz 2001:63. http://unfccc.int/resource/iuckit/ccki
t2001en.pdf [accessed 26.12.15].
[2] Trivedi J, Aila M, Bangwal DP, Kaul S, Garg MO. Algae based biorenery
How to make sense? Renew Sustain Energy Rev 2015;47:295307. http://dx.
doi.org/10.1016/j.rser.2015.03.052.
[3] Speranza LG, Ingram A, Leeke GA. Assessment of algae biodiesel viability
based on the area requirement in the European Union, United States and
Brazil. Renew Energy 2015;78:40617. http://dx.doi.org/10.1016/j.
renene.2014.12.059.
[4] Yamamoto S, Mandokoro Y, Nagano S, Nagakubo M, Atsumi K, Watanabe
MM. Catalytic conversion of Botryococcus braunii oil to diesel fuel under mild
reaction conditions. J Appl Phycol 2013;26:5564. http://dx.doi.org/10.1007/
s10811-013-0065-9.
[5] Srivastava A, Prasad R. Triglycerides-based diesel fuels. Renew Sustain En-
ergy Rev 2000;4:11133. http://dx.doi.org/10.1016/S1364-0321(99)00013-1.
[6] Pischinger GM, Falcon AM, Siekmann RW, Fernandes FR. Methylesters of
plant oils as diesels fuels, either straight or in blends. Veg Oil Fuels
1982;82:198208.
268 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
[7] Schwab AW, Bagby MO, Freedman B. Preparation and properties of diesel
fuels from vegetable oils. Fuel 1987;66:13728. http://dx.doi.org/10.1016/
0016-2361(87)90184-0.
[8] Lassing M, Mrtensson P, Olsson E, Svensson M. Biodiesel Production from
Microalgae-A Feasibility Study. Oslo, Norway; 2008.
[9] Sheehan J, Dunahay T, Benemann J, Roessler P. Look Back at the U.S. De-
partment of Energy's Aquatic Species Program: Biodiesel from Algae; Close-
Out Report. Golden, CO; 1998. http://dx.doi.org/10.2172/15003040.
[10] Nascimento IA, Marques SSI, Cabanelas ITD, Pereira SA, Druzian JI, de Souza
CO, et al. Screening microalgae strains for biodiesel production: lipid pro-
ductivity and estimation of fuel quality based on fatty acids proles as se-
lective criteria. BioEnergy Res 2013;6:113. http://dx.doi.org/10.1007/
s12155-012-9222-2.
[11] Scott SA, Davey MP, Dennis JS, Horst I, Howe CJ, Lea-Smith DJ, et al. Biodiesel
from algae: challenges and prospects. Curr Opin Biotechnol 2010;21:27786.
http://dx.doi.org/10.1016/j.copbio.2010.03.005.
[12] Lardon L, Helias A, Sialve B, Steyer J-P, Bernard O. Life-cycle assessment of
biodiesel production from microalgae. Environ Sci Technol 2009;43:6475
81. http://dx.doi.org/10.1021/es900705j.
[13] Bharathiraja B, Chakravarthy M, Ranjith Kumar R, Yogendran D, Yuvaraj D,
Jayamuthunagai J, et al. Aquatic biomass (algae) as a future feed stock for
bio-reneries: a review on cultivation, processing and products. Renew
Sustain Energy Rev 2015;47:63453. http://dx.doi.org/10.1016/j.
rser.2015.03.047.
[14] Rashid N, Ur Rehman MS, Sadiq M, Mahmood T, Han JI. Current status, issues
and developments in microalgae derived biodiesel production. Renew Sus-
tain Energy Rev 2014;40:76078. http://dx.doi.org/10.1016/j.rser.2014.07.104.
[15] Liu J, Sun Z, Gerken H. Part - III Microalgae and Bio-energy. In: Liu J, Sun Z,
Gerken H, editors. Recent Adv. Microalgal Biotechnol. Recent Adv. Microalgal
Biotechnol. Foster City: OMICS Group eBooks; 2014. p. 00318 First.
[16] Metzger P, Largeau C. Botryococcus braunii: a rich source for hydrocarbons
and related ether lipids. Appl Microbiol Biotechnol 2005;66:48696. http:
//dx.doi.org/10.1007/s00253-004-1779-z.
[17] Banerjee A, Sharma R, Chisti Y, Banerjee UC. Botryococcus braunii: a renew-
able source of hydrocarbons and other chemicals. Crit Rev Biotechnol
2002;22:24579. http://dx.doi.org/10.1080/07388550290789513.
[18] Kalacheva GS, Zhila NO, Volova TG. Lipid and hydrocarbon compositions of a
collection strain and a wild sample of the green microalga Botryococcus.
Aquat Ecol 2002;36:31730.
[19] Kalacheva GS, Zhila NO, Volova TG. Lipids of the green alga Botryococcus
cultured in a batch mode. Microbiology 2001;70:25662.
[20] Ozkan A, Kinney K, Katz L, Berberoglu H. Reduction of water and energy
requirement of algae cultivation using an algae biolm photobioreactor.
Bioresour Technol 2012;114:5428. http://dx.doi.org/10.1016/j.
biortech.2012.03.055.
[21] Bazaes J, Sepulveda C, Acin FG, Morales J, Gonzales L, Rivas M, et al. Out-
door pilot-scale production of Botryococcus braunii in panel reactors. J Appl
Phycol 2012;24:135360. http://dx.doi.org/10.1007/s10811-012-9787-3.
[22] Tran NH, Bartlett JR, Kannangara GSK, Milev S, Volk H, Wilson MA. Catalytic
upgrading of biorenery oil from micro-algae. Fuel 2010;89:26574. http:
//dx.doi.org/10.1016/j.fuel.2009.08.015.
[23] Ranga Rao A, Dayananda C, Sarada R, Shamala TR, Ravishankar G a. Effect of
salinity on growth of green alga Botryococcus braunii and its constituents.
Bioresour Technol 2007;98:5604. http://dx.doi.org/10.1016/j.
biortech.2006.02.007.
[24] Ranga Rao A, Ravishankar G a, Sarada R. Cultivation of green alga Bo-
tryococcus braunii in raceway, circular ponds under outdoor conditions and
its growth, hydrocarbon production. Bioresour Technol 2012;123:52833.
http://dx.doi.org/10.1016/j.biortech.2012.07.009.
[25] Choi GG, Kim BH, Ahn CY, Oh HM. Effect of nitrogen limitation on oleic acid
biosynthesis in Botryococcus braunii. J Appl Phycol 2011;23:10317. http:
//dx.doi.org/10.1007/s10811-010-9636-1.
[26] Lee SJ, Kim S, Kim J, Kwon G, Yoon B, Oh H. Effects of harvesting method and
growth stage on the occulation of the green alga Botryococcus braunii. Lett
Appl Microbiol 1998;27:148. http://dx.doi.org/10.1046/
j.1472-765X.1998.00375.x.
[27] Vazquez-duhalt R, Arredondo-vega B. Haloadaptation of the green alga Bo-
tryococcus braunii (race A). Phytochemistry 1991;30:291925.
[28] Yoo C, Jun SY, Lee JY, Ahn CY, Oh HM. Selection of microalgae for lipid
production under high levels carbon dioxide. Bioresour Technol 2010;101
(Suppl):S714. http://dx.doi.org/10.1016/j.biortech.2009.03.030.
[29] Cheng P, Ji B, Gao L, Zhang W, Wang J, Liu T. The growth, lipid and hydro-
carbon production of Botryococcus braunii with attached cultivation. Bior-
esour Technol 2013;138:95100. http://dx.doi.org/10.1016/j.
biortech.2013.03.150.
[30] Cheng P, Wang J, Liu T. Effects of nitrogen source and nitrogen supply model
on the growth and hydrocarbon accumulation of immobilized biolm cul-
tivation of B. braunii. Bioresour Technol 2014;166:52733. http://dx.doi.org/
10.1016/j.biortech.2014.05.045.
[31] Wang SK, Wang F, Stiles AR, Guo C, Liu CZ. Botryococcus braunii cells: ul-
trasound-intensied outdoor cultivation integrated with in situ magnetic
separation. Bioresour Technol 2014;167:37682. http://dx.doi.org/10.1016/j.
biortech.2014.06.028.
[32] Kalacheva G, Zhila N, Volova T, Gladyshev M. The effect of temperature on
the lipid composition of the green alga Botryococcus. Microbiology
2002;71:28693.
[33] Volova TG, Kalacheva GS, Zhila NO. Specicity of lipid composition in two
Botryococcus strains, the producers of liquid hydrocarbons. Russ J Plant
Physiol 2003;50:62733.
[34] Zhila NO, Kalacheva GS, Volova TG. Structure of hydrocarbons synthesized by
the alga Botryococcus isolated from Lake Shira. Dokl Biol Sci 2001;378:7037.
[35] Zhila NO, Kalacheva GS, Volova TG. Inuence of nitrogen deciency on bio-
chemical composition of the green alga Botryococcus. J Appl Phycol
2005;17:30915. http://dx.doi.org/10.1007/s10811-005-7212-x.
[36] Zhila NO, Kalacheva GS, Volova TG. Effect of salinity on the biochemical
composition of the alga Botryococcus braunii Ktz IPPAS H-252. J Appl Phycol
2011;23:4752. http://dx.doi.org/10.1007/s10811-010-9532-8.
[37] Ashokkumar V, Rengasamy R. Mass culture of Botryococcus braunii Kutz.
under open raceway pond for biofuel production. Bioresour Technol
2012;104:3949. http://dx.doi.org/10.1016/j.biortech.2011.10.093.
[38] Ashokkumar V, Rengasamy R, Deepalakshmi S, Sivalingam a, Sivakumar P.
Mass cultivation of microalgae and extraction of total hydrocarbons: a ki-
netic and thermodynamic study. Fuel 2014;119:30812. http://dx.doi.org/
10.1016/j.fuel.2013.11.062.
[39] Sydney EB, da Silva TE, Tokarski A, Novak AC, de Carvalho JC, Woiciecohwski
AL, et al. Screening of microalgae with potential for biodiesel production and
nutrient removal from treated domestic sewage. Appl Energy 2011;88:3291
4. http://dx.doi.org/10.1016/j.apenergy.2010.11.024.
[40] rpez R, Martnez ME, Hodaifa G, El Yous F, Jbari N, Snchez S. Growth of
the microalga Botryococcus braunii in secondarily treated sewage. Desalina-
tion 2009;246:62530. http://dx.doi.org/10.1016/j.desal.2008.07.016.
[41] Metzger P, Villarreal-Rosales E, Casadevall E. Methyl-branched fatty alde-
hydes and fatty acids in Botryococcus braunii. Phytochemistry 1991;30:185
91. http://dx.doi.org/10.1016/0031-9422(91)84121-8.
[42] Yamaguchi K, Nakano H, Murakami M, Konosu S, Nakayama O, Kanda M,
et al. Lipid composition of a green alga Botryococcus braunii. Agric Biol Chem
1987;51:4938.
[43] Chinnasamy S, Bhatnagar A, Hunt RW, Das KC. Microalgae cultivation in a
wastewater dominated by carpet mill efuents for biofuel applications.
Bioresour Technol 2010;101:3097105. http://dx.doi.org/10.1016/j.
biortech.2009.12.026.
[44] Yeesang C, Cheirsilp B. Low-cost production of green microalga Botryococcus
braunii biomass with high lipid content through mixotrophic and photo-
autotrophic cultivation. Appl Biochem Biotechnol 2014;174:11629. http:
//dx.doi.org/10.1007/s12010-014-1041-9.
[45] Talukdar J, Kalita MC, Goswami BC. Characterization of the biofuel potential
of a newly isolated strain of the microalga Botryococcus braunii Ktzing from
Assam, India. Bioresour Technol 2013;149:26875. http://dx.doi.org/10.1016/
j.biortech.2013.09.057.
[46] Hirose M, Mukaida F, Okada S, Noguchi T. Active hydrocarbon biosynthesis
and accumulation in a green alga, Botryococcus braunii (race A). Eukaryot Cell
2013;12:113241. http://dx.doi.org/10.1128/EC.00088-13.
[47] Grice K, Schouten S, Nissenbaum A, Charrach J, Damste JSS. A remarkable
paradox: sulfurised freshwater algal (Botryococcus braunii) lipids in an an-
cient hypersaline euxinic ecosystem. Org Geochem 1998;28:195216.
[48] Niitsu R, Kanazashi M, Matsuwaki I, Ikegami Y, Tanoi T, Kawachi M, et al.
Changes in the hydrocarbon-synthesizing activity during growth of Bo-
tryococcus braunii B70. Bioresour Technol 2012;109:2979. http://dx.doi.org/
10.1016/j.biortech.2011.08.072.
[49] Weiss TL, Chun HJ, Okada S, Vitha S, Holzenburg A, Laane J, et al. Raman
spectroscopy analysis of botryococcene hydrocarbons from the green mi-
croalga Botryococcus braunii. J Biol Chem 2010;285:3245866. http://dx.doi.
org/10.1074/jbc.M110.157230.
[50] Chiang IZ, Huang WY, Wu JT. Allelochemicals of Botryococcus braunii
(Chlorophyceae). J Phycol 2004;40:47480. http://dx.doi.org/10.1111/
j.1529-8817.2004.03096.x.
[51] Papa RD, Wu J, Baldia S, Cho C, Cruz MA, Saguiguit A, et al. Blooms of the
colonial green algae, Botryococcus braunii Ktzing, in Paoay lake, Luzon Is-
land, Philippines. Philipp J Syst Biol 2008;2:2131.
[52] Ioki M, Ohkoshi M, Nakajima N, Nakahira-Yanaka Y, Watanabe MM. Isolation
of herbicide-resistant mutants of Botryococcus braunii. Bioresour Technol
2012;109:3003. http://dx.doi.org/10.1016/j.biortech.2011.07.101.
[53] Song L, Qin JG, Clarke S, Li Y. Competition and succession between the oily
alga Botryococcus braunii and two green algae Chlorella vulgaris and Chla-
mydomonas reinhardtii. J Appl Phycol 2012;25:84753. http://dx.doi.org/
10.1007/s10811-012-9940-z.
[54] Rivas MO, Vargas P, Riquelme CE. Interactions of Botryococcus braunii cul-
tures with bacterial biolms. Microb Ecol 2010;60:62835. http://dx.doi.org/
10.1007/s00248-010-9686-6.
[55] Daz Bayona KC, Garcs LA. Effect of different media on exopolysaccharide
and biomass production by the green microalga Botryococcus braunii. J Appl
Phycol 2014;26:208795. http://dx.doi.org/10.1007/s10811-014-0242-5.
[56] Tran HL, Kwon JS, Kim ZH, Oh Y, Lee CG. Statistical optimization of culture
media for growth and lipid production of Botryococcus braunii LB572. Bio-
technol Bioprocess Eng 2010;15:27784. http://dx.doi.org/10.1007/
s12257-009-0127-7.
[57] Yeesang C, Cheirsilp B. Effect of nitrogen, salt, and iron content in the growth
medium and light intensity on lipid production by microalgae isolated from
freshwater sources in Thailand. Bioresour Technol 2011;102:303440. http:
//dx.doi.org/10.1016/j.biortech.2010.10.013.
[58] Nagaraja YP, Biradar C, Manasa KS, Venkatesh HS. Production of biofuel by
using micro algae (Botryococcus braunii) sample collection. Int J Curr
 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
Micobiol Appl Sci 2014;3:85160.
[59] Sushchik NN, Kalacheva GS, Zhila NO, Gladyshev MI, Volova TG. A tem-
perature dependence of the intra- and extracellular fatty-acid composition
of green algae and Cyanobacterium. Russ J Plant Physiol 2003;50:37480.
[60] Bjornsson WJ, MacDougall KM, Melanson JE, OLeary SJB, McGinn PJ. Pilot-
scale supercritical carbon dioxide extractions for the recovery of triacylgly-
cerols from microalgae: a practical tool for algal biofuels research. J Appl
Phycol 2011;24:54755. http://dx.doi.org/10.1007/s10811-011-9756-2.
[61] Cheng P, Wang J, Liu T. Effect of cobalt enrichment on growth and hydro-
carbon accumulation of Botryococcus braunii with immobilized biolm at-
tached cultivation. Bioresour Technol 2015;177:2048. http://dx.doi.org/
10.1016/j.biortech.2014.11.088.
[62] Manchanda T, Tyagi R, Sharma DK, Gautam K. Application of sea water for
hydrocarbon and oil production by Botryococcus braunii. Adv Sci Lett
2014;20:4.
[63] Tyagi R, Manchanda T, Bhardawaj A, Bajpai RB. Application of NaCl for cul-
tivation of isolated Botryococcus braunii strains. Int J Agric Food Sci Technol
2013;4:22732.
[64] Ruangsomboon S. Effect of light, nutrient, cultivation time and salinity on
lipid production of newly isolated strain of the green microalga, Botryococcus
braunii KMITL 2. Bioresour Technol 2012;109:2615. http://dx.doi.org/
10.1016/j.biortech.2011.07.025.
[65] Ranga Rao A, Sarada R, Ravishankar GA. Inuence of CO2 on growth and
hydrocarbon production in Botryococcus braunii. J Microbiol Biotechnol
2007;17:4149.
[66] Yang S, Wang J, Cong W, Cai Z, Ouyang F. Effects of bisulte and sulte on the
microalga Botryococcus braunii. Enzyme Microb Technol 2004;35:4650.
http://dx.doi.org/10.1016/j.enzmictec.2004.03.014.
[67] Wu LF, Chen PC, Lee CM. The effects of nitrogen sources and temperature on
cell growth and lipid accumulation of microalgae. Int Biodeterior Biodegrad
2013;85:50610. http://dx.doi.org/10.1016/j.ibiod.2013.05.016.
[68] Xu L, Wang F, Guo C, Liu CZ. Improved algal oil production from Botryococcus
braunii by feeding nitrate and phosphate in an airlift bioreactor. Eng Life Sci
2012;12:1717. http://dx.doi.org/10.1002/elsc.201100110.
[69] Pooja K, Himabindu V. Mixotrophic cultivation of Botryococcus braunii for
biomass and lipid yields with simultaneous CO2 sequestration. Int J Eng Res
Appl 2014;4:1516.
[70] Dayananda C, Sarada R. Inuence of nitrogen sources on growth, hydro-
carbon and fatty acid production by Botryococcus braunii. Asian J Plant Sci
2006;5:799804.
[71] Ge Y, Liu J, Tian G. Growth characteristics of Botryococcus braunii 765 under
high CO2 concentration in photobioreactor. Bioresour Technol
2011;102:1304. http://dx.doi.org/10.1016/j.biortech.2010.06.051.
[72] Okumura C, Saffreena N, Rahman MA, Hasegawa H. Economic efciency of
different light wavelengths and intensities using LEDs for the cultivation of
green microalga Botryococcus braunii (NIES-836) for biofuel production.
Environ Prog Sustain Energy 2014;00:17. http://dx.doi.org/10.1002/ep.
[73] Razzak SA, Hossain MM, Lucky RA, Bassi AS, de Lasa H. Integrated CO2
capture, wastewater treatment and biofuel production by microalgae cul-
turingA review. Renew Sustain Energy Rev 2013;27:62253. http://dx.doi.
org/10.1016/j.rser.2013.05.063.
[74] Zelibor JL, Romankiw L, Hatcher PG, Colwell RR. Comparative analysis of the
chemical composition of mixed and pure cultures of green algae and their
decomposed residues by 13C nuclear magnetic resonance spectroscopy. Appl
Environ Microbiol 1988;54:105160.
[75] Cai T, Park SY, Li Y. Nutrient recovery from wastewater streams by micro-
algae: status and prospects. Renew Sustain Energy Rev 2013;19:3609. http:
//dx.doi.org/10.1016/j.rser.2012.11.030.
[76] McGinn PJ, Dickinson KE, Bhatti S, Frigon JC, Guiot SR, OLeary SJB. Integra-
tion of microalgae cultivation with industrial waste remediation for biofuel
and bioenergy production: opportunities and limitations. Photosynth Res
2011;109:23147. http://dx.doi.org/10.1007/s11120-011-9638-0.
[77] Rodol L, Chini Zittelli G, Bassi N, Padovani G, Biondi N, Bonini G, et al.
Microalgae for oil: strain selection, induction of lipid synthesis and outdoor
mass cultivation in a low-cost photobioreactor. Biotechnol Bioeng
2009;102:10012. http://dx.doi.org/10.1002/bit.22033.
[78] Honda R, Boonnorat J, Chiemchaisri C, Chiemchaisri W, Yamamoto K. Carbon
dioxide capture and nutrients removal utilizing treated sewage by con-
centrated microalgae cultivation in a membrane photobioreactor. Bioresour
Technol 2012;125:5964. http://dx.doi.org/10.1016/j.biortech.2012.08.138.
[79] Wiley P, Harris L, Reinsch S, Tozzi S, Embaye T, Clark K, et al. Microalgae
cultivation using offshore membrane enclosures for growing algae (OMEGA).
J Sustain Bioenergy Syst 2013;03:1832. http://dx.doi.org/10.4236/
jsbs.2013.31003.
[80] Brennan L, Owende P. Biofuels from microalgaeA review of technologies
for production, processing, and extractions of biofuels and co-products. Re-
new Sustain Energy Rev 2010;14:55777. http://dx.doi.org/10.1016/j.
rser.2009.10.009.
[81] Lam MK, Lee KT. Microalgae biofuels: a critical review of issues, problems
and the way forward. Biotechnol Adv 2012;30:67390. http://dx.doi.org/
10.1016/j.biotechadv.2011.11.008.
[82] Chen CY, Yeh KL, Aisyah R, Lee DJ, Chang JS. Cultivation, photobioreactor
design and harvesting of microalgae for biodiesel production: a critical re-
view. Bioresour Technol 2011;102:7181. http://dx.doi.org/10.1016/j.
biortech.2010.06.159.
[83] Rawat I, Ranjith Kumar R, Mutanda T, Bux F. Biodiesel from microalgae: a
269
critical evaluation from laboratory to large scale production. Appl Energy
2013;103:44467. http://dx.doi.org/10.1016/j.apenergy.2012.10.004.
[84] Eroglu E, Okada S, Melis A. Hydrocarbon productivities in different Bo-
tryococcus strains: comparative methods in product quantication. J Appl
Phycol 2011;23:76375. http://dx.doi.org/10.1007/s10811-010-9577-8.
[85] Kim DG, Oh HM, Park YH, Kim HS, Lee HG, Ahn CY. Optimization of occu-
lation conditions for Botryococcus braunii using response surface metho-
dology. J Appl Phycol 2012;25:87582. http://dx.doi.org/10.1007/
s10811-012-9948-4.
[86] Xu L, Wang F, Li HZ, Hu ZM, Guo C, Liu CZ. Development of an efcient
electroocculation technology integrated with dispersed-air otation for
harvesting microalgae. J Chem Technol Biotechnol 2010;85:15047. http:
//dx.doi.org/10.1002/jctb.2457.
[87] Lee SJ, Yoon B, Oh H. Rapid method for the determination of lipid from the
green alga Botryococcus braunii. Biotechnol Tech 1998;12:5536.
[88] Lee AK, Lewis DM, Ashman PJ. Disruption of microalgal cells for the ex-
traction of lipids for biofuels: processes and specic energy requirements.
Biomass Bioenergy 2012;46:89101. http://dx.doi.org/10.1016/j.
biombioe.2012.06.034.
[89] McNichol J, MacDougall KM, Melanson JE, McGinn PJ. Suitability of Soxhlet
extraction to quantify microalgal fatty acids as determined by comparison
with in situ transesterication. Lipids 2012;47:195207. http://dx.doi.org/
10.1007/s11745-011-3624-3.
[90] Moheimani NR, Cord-Ruwisch R, Raes E, Borowitzka M. a. Non-destructive
oil extraction from Botryococcus braunii (Chlorophyta). J Appl Phycol
2013;25:165361. http://dx.doi.org/10.1007/s10811-013-0012-9.
[91] Sawayama S, Minowa T, Yokoyama SY. Possibility of renewable energy pro-
duction and CO2 mitigation by thermochemical liquefaction of microalgae.
Biomass Bioenergy 1999;17:339. http://dx.doi.org/10.1016/S0961-9534(99)
00019-7.
[92] Heger M. A new processing scheme for algae biofuels. MIT Technol Rev 2009.
http://www.technologyreview.com/news/413325/a-new-processing-
scheme-for-algae-biofuels/.
[93] Halim R, Danquah MK, Webley P a. Extraction of oil from microalgae for
biodiesel production: a review. Biotechnol Adv 2012;30:70932. http://dx.
doi.org/10.1016/j.biotechadv.2012.01.001.
[94] Mubarak M, Shaija A, Suchithra TV. A review on the extraction of lipid from
microalgae for biodiesel production. Algal Res 2014. http://dx.doi.org/
10.1016/j.algal.2014.10.008.
[95] Santana A, Jesus S, Larrayoz MA, Filho RM. Supercritical carbon dioxide ex-
traction of algal lipids for the biodiesel production. Proc Eng 2012;42:1755
61. http://dx.doi.org/10.1016/j.proeng.2012.07.569.
[96] Shi F, Wang P, Duan Y, Link D, Morreale B. Recent developments in the
production of liquid fuels via catalytic conversion of microalgae: experi-
ments and simulations. RSC Adv 2012;2:972747. http://dx.doi.org/10.1039/
c2ra21594b.
[97] Pragya N, Pandey KK, Sahoo PK. A review on harvesting, oil extraction and
biofuels production technologies from microalgae. Renew Sustain Energy
Rev 2013;24:15971. http://dx.doi.org/10.1016/j.rser.2013.03.034.
[98] Kim J, Yoo G, Lee H, Lim J, Kim K, Kim CW, et al. Methods of downstream
processing for the production of biodiesel from microalgae. Biotechnol Adv
2013;31:86276. http://dx.doi.org/10.1016/j.biotechadv.2013.04.006.
[99] Petchmala A, Laosiripojana N, Jongsomjit B, Goto M, Panpranot J, Mekasu-
wandumrong O, et al. Transesterication of palm oil and esterication of
palm fatty acid in near- and super-critical methanol with SO4ZrO2 cata-
lysts. Fuel 2010;89:238792. http://dx.doi.org/10.1016/j.fuel.2010.04.010.
[100] Szczsna Antczak M, Kubiak A, Antczak T, Bielecki S. Enzymatic biodiesel
synthesis Key factors affecting efciency of the process. Renew Energy
2009;34:118594. http://dx.doi.org/10.1016/j.renene.2008.11.013.
[101] Hidalgo P, Toro C, Ciudad G, Schober S, Mittelbach M, Navia R. Evaluation of
different operational strategies for biodiesel production by direct transes-
terication of microalgal biomass. Energy Fuels 2014;28:381420.
[102] Levine RB, Pinnarat T, Savage PE. Biodiesel production from wet algal bio-
mass through in situ lipid hydrolysis and supercritical transesterication.
Energy Fuels 2010;24:523543. http://dx.doi.org/10.1021/ef1008314.
[103] Atadashi IM, Aroua MK, Aziz a A. Biodiesel separation and purication: a
review. Renew Energy 2011;36:43743. http://dx.doi.org/10.1016/j.
renene.2010.07.019.
[104] Atadashi IM, Aroua MK, Aziz a RA, Sulaiman NMN. Rening technologies for
the purication of crude biodiesel. Appl Energy 2011;88:423951. http://dx.
doi.org/10.1016/j.apenergy.2011.05.029.
[105] Atadashi IM, Aroua MK, Abdul Aziz a R, Sulaiman NMN. Membrane biodiesel
production and rening technology: a critical review. Renew Sustain Energy
Rev 2011;15:505162. http://dx.doi.org/10.1016/j.rser.2011.07.051.
[106] Atadashi IM, Aroua MK, Aziz AA. High quality biodiesel and its diesel engine
application: a review. Renew Sustain Energy Rev 2010;14:19992008. http:
//dx.doi.org/10.1016/j.rser.2010.03.020.
[107] Rawat I, Ranjith Kumar R, Mutanda T, Bux F. Dual role of microalgae: phy-
coremediation of domestic wastewater and biomass production for sustain-
able biofuels production. Appl Energy 2011;88:341124. http://dx.doi.org/
10.1016/j.apenergy.2010.11.025.
[108] Li YG, Xu L, Huang YM, Wang F, Guo C, Liu CZ. Microalgal biodiesel in China:
opportunities and challenges. Appl Energy 2011;88:34327. http://dx.doi.
org/10.1016/j.apenergy.2010.12.067.
[109] Yang Z, Guo R, Xu X, Fan X, Luo S. Fermentative hydrogen production from
lipid-extracted microalgal biomass residues. Appl Energy 2011;88:346872.
270 M.B. Tasi et al. / Renewable and Sustainable Energy Reviews 64 (2016) 260270
http://dx.doi.org/10.1016/j.apenergy.2010.09.009.
[110] Watanabe H, Li D, Nakagawa Y, Tomishige K, Kaya K, Watanabe MM. Char-
acterization of oil-extracted residue biomass of Botryococcus braunii as a
biofuel feedstock and its pyrolytic behavior. Appl Energy 2014;132:47584.
http://dx.doi.org/10.1016/j.apenergy.2014.07.037.
[111] Cabanelas ITD, Marques SSI, de Souza CO, Druzian JI, Nascimento IA. Bo-
tryococcus, what to do with it? Effect of nutrient concentration on bior-
enery potential Algal Res 2015;11:439. http://dx.doi.org/10.1016/j.
algal.2015.05.009.
[112] Ruangsomboon S. Effects of different media and nitrogen sources and levels
on growth and lipid of green microalga Botryococcus braunii KMITL and its
biodiesel properties based on fatty acid composition. Bioresour Technol
2015;191:37784. http://dx.doi.org/10.1016/j.biortech.2015.01.091.
[113] Jones CS, Mayeld SP. Algae biofuels: versatility for the future of bioenergy.
Curr Opin Biotechnol 2012;23:34651. http://dx.doi.org/10.1016/j.
copbio.2011.10.013.
[114] Thurmond W. Algae 2020: Advanced Biofuel Markets and Commercialization
Outlook. rst ed. Emerging Markets Online; 2009.
[115] Chisti Y. Biodiesel from microalgae. Biotechnol Adv 2007;25:294306. http:
//dx.doi.org/10.1016/j.biotechadv.2007.02.001.
[116] Kirrolia A, Bishnoi NR, Singh R. Microalgae as a boon for sustainable energy
production and its future research & development aspects. Renew Sustain
Energy Rev 2013;20:64256. http://dx.doi.org/10.1016/j.rser.2012.12.003.
[117] Singh J, Gu S. Commercialization potential of microalgae for biofuels pro-
duction. Renew Sustain Energy Rev 2010;14:2596610. http://dx.doi.org/
10.1016/j.rser.2010.06.014.
[118] Davis R, Aden A, Pienkos PT. Techno-economic analysis of autotrophic mi-
croalgae for fuel production. Appl Energy 2011;88:352431. http://dx.doi.org/
10.1016/j.apenergy.2011.04.018.
[119] Woertz IC, Benemann JR, Du N, Unnasch S, Mendola D, Mitchell BG, et al. Life
cycle GHG emissions from microalgal biodiesela CA-GREET model. Environ
Sci Technol 2014;48:60608. http://dx.doi.org/10.1021/es403768q.
[120] Zaimes GG, Khanna V. Environmental sustainability of emerging algal bio-
fuels : a comparative life cycle evaluation of algal biodiesel and renewable
diesel. Environ Prog Sustain Energy 2013;00:15. http://dx.doi.org/10.1002/
ep.
[121] Stephenson AL, Kazamia E, Dennis JS, Howe CJ, Scott S a, Smith AG. Life-cycle
assessment of potential algal biodiesel production in the united kingdom: a
comparison of raceways and air-lift tubular bioreactors. Energy Fuels
2010;24:406277. http://dx.doi.org/10.1021/ef1003123.
[122] Frank ED, Elgowainy A, Han J, Wang Z. Life cycle comparison of hydrothermal
liquefaction and lipid extraction pathways to renewable diesel from algae.
Mitig Adapt Strateg Glob Change 2012;18:13758. http://dx.doi.org/10.1007/
s11027-012-9395-1.
[123] Gutirrez-Arriaga CG, Serna-Gonzlez M, Ponce-Ortega JM, El-Halwagi MM.
Sustainable integration of algal biodiesel production with steam electric
power plants for greenhouse gas mitigation. ACS Sustain Chem Eng
2014;2:1388403. http://dx.doi.org/10.1021/sc400436a.
[124] Shiho M, Kawachi M, Horioka K, Nishita Y, Ohashi K, Kaya K, et al. Business
evaluation of a green microalgae Botryococcus braunii oil production system.
Proc Environ Sci 2012;15:90109. http://dx.doi.org/10.1016/j.
proenv.2012.05.014.