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Renewable and Sustainable Energy Reviews: Emmanuel B. D 'Alessandro, Nelson R. Antoniosi Filho
Renewable and Sustainable Energy Reviews: Emmanuel B. D 'Alessandro, Nelson R. Antoniosi Filho
art ic l e i nf o a b s t r a c t
Article history: This review describes compounds produced microalgae, such as biodiesel, lipids, fatty acids (FA), tria-
Received 11 December 2015 cylglycerides (TAG), and pigments (phycobilins, chlorophylls, and carotenoids). We discuss the factors
Accepted 17 December 2015 inducing the accumulation of these metabolites and their economic importance. We focused on cell wall
breaking methods of microalgae used to produce biodiesel. A special approach was made to extremo-
Keywords: phile microalgae used in biodiesel production. The type of methodology used in the cultivation and the
Biodiesel use of extremophiles microalgae can permit feasible biodiesel production.
Carotenoids & 2016 Elsevier Ltd. All rights reserved.
Stress
Extremophile
Lipid metabolism
Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 832
2. Lipids of microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 833
3. Fatty acids in microalgae. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 833
4. Factors influencing the increase of lipids in microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 834
5. Pigments in microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 835
5.1. Phycobilins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 835
5.2. Chlorophyll. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 835
5.3. Carotenoids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 835
6. Market for pigment production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 836
7. Cell membrane of microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 837
8. Extremophile microalgae . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 837
9. Metabolism of fatty acids and acylglycerides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 838
10. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 838
Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 839
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 839
http://dx.doi.org/10.1016/j.rser.2015.12.162
1364-0321/& 2016 Elsevier Ltd. All rights reserved.
E.B. D’Alessandro, N.R. Antoniosi Filho / Renewable and Sustainable Energy Reviews 58 (2016) 832–841 833
vegetable oils for their uses as fuels" [4]. Biodiesel production is Table 1
mainly based on grease raw materials, such as palm oil, canola oil, Lipid content in different cyanobacteria and microalgae.
animal tallow, and soybean oil [5]. However, the prices of these
Division Lipid content (%) Reference
commodities are regulated internationally and affect biodiesel
price [6]. For example, about 80% of biodiesel cost vary with the Cyanophyta
price of grease raw materials [7], suggesting that new greases Lyngbya birgei 127 2.8 [22]
sources are needed in order to reduce the biodiesel price. Spirulina maxima 67 [10]
Spirulina platensis 8.5 7 2 [22]
As a result, there is growing interest in using microalgae for Spirulina platensis 4–9 [10]
biodiesel production, especially because some produce about 150 Synechocystis pevalekii 97 2 [22]
times more oil per hectare than soybean [8]. Soybean is the main Chlorophyta
oilseed used for biodiesel production in Brazil. However, turning Ankistrodesmus falcatus 1.58 [23]
Ankistrodesmus gracilis 7.9–20.5 [24]
biodiesel production from microalgae into an economically viable
Auxenochlorella protothecoides 39.3 7 0.8 [25]
activity is still a challenge [9]. Auxenochlorella protothecoides 32.9 7 0.6 [25]
Producing biofuel from microalgae has many advantages [10], Botryococcus braunii 14.0–28.6 [26]
such as: (1) high growth rate; (2) lower demand for water than Botryococcus braunii 25–75 [19]
commercial crops; (3) High efficiency in CO2 mitigation; (4) smal- Chlamydomonas rheinhardii 21 [10]
Chlorella pyrenoidosa 2 [10]
ler areas for cultivation. However, there are also disadvantages, Chlorella sorokiniana 19 [27]
such as: (1) dependence of light incidence and penetration into Chlorella spp. 10.5 [23]
the aquatic environment to ensure high biomass production Chlorella vulgaris 12.0 7 0.56 [28]
(2) difficulty of developing simple and inexpensive procedures to Chlorella vulgaris 267 0.5 [25]
Chlorella vulgaris 14 22 [10]
convert lipids into biodiesel.
Chlorococcum infusionum 11.3 7 1 [22]
Additionally, there are other drawbacks related to the selection Dunaliella bioculata 8 [10]
of species with appropriate production time and yield of biomass, Dunaliella salina 6 [10]
lipids, and pigments. Ideal species would also have biomass easily Scenedesmus acuminatus 1.58 [23]
separable from the culture medium, adapted to low-cost cultiva- Scenedesmus dimorphus 16–40 [10]
Scenedesmus obliquus 12–14 [10]
tion conditions, and resistant to invasive organisms. Scenedesmus obliquus 6.187 0.20 [29]
Thus, closed systems (photobioreactors) and open ponds Scenedesmus quadricauda 1.9 [10]
(raceways) are used for microalgae cultivation on a large scale and Spirogyra sp. 11-21 [10]
both have advantages and disadvantages. The cost to build and Spirogyra orientalis 217 2.5 [22]
Heterokontophyta
operate a closed system is higher than ponds. Conversely, it
Navicula minima 16.2 7 0.6 [22]
requires less light and area, besides having volumetric productivity Nitzschia spp 3.68 [23]
about 13 times higher than raceways [11]. Phaeodactylum tricornutum 9.40 7 1.77 [28]
Key aspects of lipid metabolism in microalgae have been ana- Euglenophyta
lyzed to improve biodiesel production. For example: (1) stress may Euglena acus 5.78 [23]
Euglena gracilis 14–20 [10]
increase lipid content [12]; (2) manipulating the growing medium Rhodophyta
can increase biomass productivity [13]; (3) some microalgae can Catenella repens 87 1.5 [22]
survive in extreme environments, which facilitates production in Ceramium manorensis 87 1.9 [22]
raceway and decreases contamination [14]; (4) the lipid metabo- Geledium pusillum 9.7 7 2.8 [22]
Dinophyta
lism is known at the molecular level ([15]; and 5) some microalgae
Gymnodinium sp. 29.6 [30]
produce valuable pigments, such as astaxanthin, lutein, and β-
carotene, which can prevent and treat diseases [16].
Thus, the cultivation of microalgae for biodiesel production The main Chlorophyta studied for biodiesel production are
involves not only the selection and production of species, assess- unicellular, such as Auxenochlorella protothecoides, Chlorella vul-
ment of types and amount of lipids, but also the potential market garis, Chlamydomonas rheinhardii, and Dunaliella salina. Since
for co-products. growth rate of these species are faster, lipid productivity is higher
than in other divisions. But the divisions Euglenophyta and
Dinophyta have potential to be used for biodiesel production, since
2. Lipids of microalgae their lipid amount is greater than some Chlorophyta (Table 1).
Cyanophyta are also evaluated for biodiesel production, due to
Lipids can be classified into two groups: polar and neutral. They their high cell growth rate. However, some produce toxic and
are insoluble in water but soluble in most organic solvents. Polar
bioaccumulative substances, such as microcystin, which is carci-
lipids include phospholipids and glycolipids, while neutral lipids
nogen [20].
include acylglicerids (tri, di- and monoglycerides) and free fatty
It is also important to assess the types of fatty acids in micro-
acids. Microalgae use neutral lipids as energy source and polar
algae, since they influence biodiesel quality, especially oxidative
lipids to form cell membranes. Nonetheless, microalgae also have
stability, cold filter plugging point, and contents of mono-, di- and
fatty-acid free components that are not converted into biodiesel,
triglycerides [21].
such as steroids and pigments [17]. As a result, higher production
of pigment implies lower production of fatty acids. Therefore,
although some microalgae produce high lipid content [18,19], this
not necessary means a high biodiesel production. Accordingly, the 3. Fatty acids in microalgae
percentage of lipid content per dry weight of microalgae may
range from 1.5% to 75% (Table 1), and a given microalga can pro- Several authors have evaluated fatty acid composition in
duce different amounts of lipids depending on the culture medium microalgae [18,30–37]. Fatty acids are carboxylic acids with 4-36
and procedure. For example, the lipid content of Chlorella vulgaris carbons chains. Fatty acids without unsaturations are called satu-
ranged from 12% to 26% and Botryococcus braunii ranges from 14% rated (SFA), those with only one unsaturated bond are called
to 75%. monounsaturated (MUFA), those with two are di-unsaturated
834 E.B. D’Alessandro, N.R. Antoniosi Filho / Renewable and Sustainable Energy Reviews 58 (2016) 832–841
Table 2 Table 5
Main saturated fatty acids (SFA) in microalgae. Main tri-unsaturated fatty acids (TUFA) in microalgae.
Fatty acids Common name Representation Fatty acids Common name Representations
Table 6
Main polyunsaturated fatty acids (PUFA) in microalgae.
cis-6, cis-9, cis-12, cis-15 octadecatetraenoic Stearidonic C18:4 c6 c9 c12 c15 or C18:4ω3
cis-5, cis-8, cis-11, cis-14 eicosatetraenoic Arachidonic C20:4 c5 c8 c11 c14 or C20:4ω6
cis-8, cis-11, cis-14, cis-17 eicosatetraenoic – C20:4 c8 c11 c14 c17 or C20:4ω3
cis-7, cis-10, cis-13, cis-16 docosatetraenoic Adrenic C22:4 c7 c10 c13 c16 or C22:4ω6
cis-10, cis-13, cis-16, cis-19 docosatetraenoic – C22:4 c10 c13 c16 c19 or C22:4ω3
cis-5, cis-8, cis-11, cis-14, cis-17 eicosapentaenoic EPA C20:5 c5 c8 c11 c14 c17 or C20:5ω3
cis-4, cis-7, cis-10, cis-13, cis-16 docosapentaenoic Docosapentaenoic C22:5 c4 c7 c10 c13 c16 or C22:5ω6
cis-7, cis-10, cis-13, cis-16, cis-19 docosapentaenoic DPA C22:5 c7 c10 c13 c16 c19 or C22:5ω3
cis-4, cis-7, cis-10, cis-13, cis-16, cis-19 docosahexaenoic DHA C22:6 c4 c7 c10 c13 c16 c19 or C22:6ω3
with increasing light intensity and concluded that the best con- number of patents, the range of red algae and cyanobacteria stu-
dition was 600 mE m2/s with lipid productivity of 85 mg/L/day. died for this purpose is still small.
Zhang et al. [52] found that Chlorella vulgaris produced more
lipid in photoautotrophic cultures mixed with bacteria than in 5.2. Chlorophyll
pure autotrophic and heterotrophic cultures. Previous studies have
also shown that mixed photoautotrophic cultures are more ben- Chlorophylls are greenish and fat-soluble pigments with a
eficial and low cost, eliminating the need to add organic carbon porphyrin ring [55]. They are responsible for converting solar
sources. Therefore, mixed photoautotrophic cultures are more energy into chemical energy in photosynthesis. Most microalgae
economically viable to produce biodiesel than autotrophic or have chlorophyll a, but some may have chlorophyll b and c (Fig. 2),
heterotrophic cultures. such as Dinophyta [58]. Microalgae contain from 0.5 to 1.0% of
chlorophyll per dry weight [57].
Chlorophyllin is a derivate of chlorophyll in which the mag-
nesium is replaced by sodium or copper and phytol chains are lost
5. Pigments in microalgae
[59]. Chlorophyllins have been used to control body odor of ger-
iatric patients [60], and dietary supplement [61]. Several studies
Natural pigments have an important role in the photosynthetic
found that chlorophyll and chlorophyllin have also antimutagenic
metabolism and pigmentation in algae. Also, they have beneficial
and anticarcinogenic action [62–65].
biological activities, such as antioxidant, anticancer, anti-inflam-
matory, anti-obesity, anti-angiogenic, and neuroprotective [53,54].
5.3. Carotenoids
At least three classes of pigments occur in microalgae: phycobilins,
chlorophylls, and carotenoids.
Carotenoids (Fig. 3) are fat-soluble substances with colors
varying form brown, red, orange to yellow. They perform two key
5.1. Phycobilins roles in photosynthesis: 1) absorb light in regions of the visible
spectrum, in which chlorophyll does not absorb efficiently; 2)
Phycobilins (or phycobiliproteins; Fig. 1) are soluble in water photoprotect the photosynthetic systems. Photoprotection
and relatively easy to isolate and purify, because they comprise a mechanisms remove the most energetic states of chlorophyll,
large portion of the total cell protein. Phycobilins are accessory for resulting from the excessive absorption of light radiation. This
collecting light during photosynthesis. For example, at four phy- hinders the formation of reactive oxygen species (ROS), makes
cobilin classes are produced in red algae (Rhodophyta and Glau- carotenoids good antioxidants [66]. The main carotenoids of
cophytas): allophycocyanin (green-bluish), phycocyanin (blue), microalgae are: β-carotene, lycopene, astaxanthin, zeaxanthin,
phycoerythrin (purple), and phycoerythrocyanin (orange) [55,56]. violaxanthin, and lutein. Of these, β-carotene, lutein, and astax-
Phycobilins are widely used in industry and immunology anthin are the most studied ones [67].
laboratories, due to their absorption properties. They are often β-carotene is a orange-yellowish pigment with growing
employed in molecular biology as fluorescent markers, used in demand, being used as colorant for food or nutritional supple-
immunoassays and as fluorescent dyes for microscopy [57]. There ment, because it is a precursor of vitamin A (retinol) [68,69].
are at least 297 patent records for these substances, mostly based Demand for natural carotenoids is increasing, because the natural
on applications, such as fluorescent markers. The United States of form has properties that the artificial one does not. For example,
America have most patents related to fluorescent applications, Jayappriyan et al. [70] found that β-carotene from Dunaliella salina
while Japan has patents relating to production, purification, and causes a higher apoptosis rate in prostate cancer cells than the
therapeutic and diagnostic purposes [56]. Despite the large synthetic version.
836 E.B. D’Alessandro, N.R. Antoniosi Filho / Renewable and Sustainable Energy Reviews 58 (2016) 832–841
microalgae is US $ 2500/kg to $ 3000/kg, at production cost of H2O2 alone and with H2O2 þFeSO4. Lipid extraction in the first
microalgae of US$ 5 to US$ 20/kg dry weight [100]. The artificial treatment increased from 6.9 to 9.2% and from 6.9 to 17.4% in the
form costs around US$ 1000/kg. However, Li et al. [85] surveyed second treatment. Authors also optimized the time that algal
the costs and concluded that China can sell astaxanthin at a lower biomass was exposed to peroxide at 4 minutes, since peroxide
price (US$ 718/kg) using low-cost photobioreactors, two phases in starts degrading lipids after that time.
raceways (ponds), and low cost of labor and reagents. Thus, it is important that the cell membrane of microalgae used
Thus, various carotenoids have high commercial value and can for biodiesel production and other byproducts (e.g., pigments) be
be produced from microalgae together with biodiesel, specially for rigid to allow movement and isolation of biomass in the culture.
their high demand as nutraceutical. Thus, antioxidant pigments, However, cell membrane cannot be too rigid, because it could
known as nutraceuticals, are currently the most marketed pro- make component extraction unviable.
ducts from microalgae, renovating the interest in increasing the
concentration of these substances in microalgae.
Stressful conditions in cultivation (e.g., changes in nutrient 8. Extremophile microalgae
levels or light intensity), induce microalgae to produce ROS as a
signaling molecule for control processes, such as cell death pro- Important parameters to select microalgae for biodiesel pro-
gramming, response to abiotic stress, pathogen defense, and sys- duction include lipid content and production, biomass productiv-
temic signaling. ROS are reduced forms of atmospheric oxygen ity and potential of adaptability to different growth conditions.
(O2), produced from O2 excitement to form singlet oxygen (1O2) or Additionally, the same microalgae species can produce different
from the transfer of one, two, or three electrons to O2, forming a fatty acids depending on the environment in which it is grown. As
superoxide radical (O2 ), hydrogen peroxide (H2O2), or hydroxyl a result, recent studies have attempted to increase biomass pro-
radical ( OH), respectively. Therefore, they can oxidize various ductivity or change the profile and content of fatty acids of
cellular components and cause oxidative cell destruction [101]. microalgae by using species collected in extreme environments, or
Thus, microalgae produce ROS as a defense mechanism to stress, collected in conventional environments, but grown in extreme
producing antioxidant enzymes as by-products, such as super- ones, such as thermal, acidic, alkaline, salty, or eutrophic waters.
oxide dismutase, catalase, peroxidase, and glutathione reductase, For example, Teoh et al. [115] compared the fatty acid profile of
as well as non-enzymatic antioxidant molecules, such as phy- Chlamydomonas from the Antarctica, temperate and tropical
tochelatins, polysaccharides, polyphenols, and pigments [102,103]. environments. The percentage of saturated fatty acids of Antarctic
However, few studies relate ROS to pigment induction in micro- Chlamydomonas increased from 21.1% when grown at 4 °C to 32.6%
algae, despite the pathways for pigment production being known. when grown at 20 °C, while that of temperate Chlamydomonas
increased from 48.7% at 4 °C to 51.6% at 25 °C. Finally, the content
of saturated fatty acids of tropical Chlamydomonas increased from
7. Cell membrane of microalgae 28.7% when grown at 18 °C to 69.8% at 33 °C. Thus, the percentage
of saturated fatty acids of microalgae from warmer environments
Several steps are involved in converting microalgae products or cultured in high temperatures may increase. This suggests that
(e.g., pigments and fatty acids) into biodiesel, specially breaking microalgae reduces the production of unsaturated fatty acids,
microalgae cell, since it influences lipid productivity and pigments. which maintains the cellular content in the liquid state at low
Studies of cell disruption have advanced in recent years [104–106]. temperatures, changing to produce saturated fatty acids that
For example, the efficiency of cell disruption methods depends on provide more energy. Accordingly, An et al. [116] found genes in
microalgae species and is related to cell membrane composition microalgae more sensitive at low temperatures, and associated
and morphology [107]. Moreover, culture medium influences cell unsaturated fatty acids production to cell protection, preventing
membrane rigidity [9]. Thus, the costs of breakage and extraction freezing. Also, Ghozzi et al. [117] collected microalgae and cyano-
can be significantly reduced by using the proper medium. bacteria from different geothermal sources in Tunisia, in which
Solvents are key components of lipid extraction, since several temperatures ranged from 40 to 61 °C. They found 14 taxa of
studies found a variation in lipid content extracted from the same cyanobacteria, but only three microalgae above 60 °C, suggesting
microalgae using different solvents and extraction forms [108– that cyanobacteria are more adapted to high temperatures than
112]. In turn, lipid content is influenced not only by the solubility microalgae.
of fatty acids, but also by solvent ability to permeate and break cell Onay et al. [118] collected Micractinium sp., H. tetrachotoma, and
membrane. Scenedesmus sp. in geothermal waters (43–44 °C) in Turkey. They
There are two methods for breaking cell membrane [107]: cultivated these species at three temperatures (25, 37, and 50 °C),
(1) the mechanical one, subdivided into grinding, high-speed and found that biomass productivity of all decay at 50 °C. Con-
homogenization, high pressure homogenization, ultrasonication, versely, biomass productivity of Micractinium sp. increased at
microwave, and pulsed electric field; and (2) non-mechanical, 37 °C, but lipid content was greater at 25 °C than 37 °C (22.7 and
subdivided into chemical and enzymatic. The mechanical method 10.7%, respectively). These results were similar to Smith-Bädorf
is the best one for industry, but its energy consumption is high. et al. [119]. Therefore, microalgae from warm environments may
Conversely, the treatment time of the non-mechanical method is be resistant to high temperatures, but generally biomass and fatty
longer, affecting product quality, although consuming less energy acid productivity is greater under conventional conditions (20–
and disrupting cell membrane uniformly. Furthermore, the non- 30 °C).
mechanical method is more difficult to control than mechanical Thus, microalgae usually produce large amounts of poly-
methods. unsaturated fatty acids, which are useless for biodiesel production,
The laser is a new mechanical method of cell disruption of because they reduce biofuel oxidative stability. As a result, using
microalgae, whereas the hydrogen peroxide (H2O2) is a new non- species adapted to or cultivated in high temperatures can provide
mechanic one. McMillan et al. [113] tested five methods for fatty acids profile more suitable for biodiesel production, i.e. with
breaking cells of N. oculata: microwave, heated bath, blender, lower proportion of unsaturated fatty acids.
ultrasound, and laser (1064 nm). Laser was the best one, with a Ruiz-Dominguez et al. [14] isolated Coccomyxa sp. from acidic
mean of 96.5% breakage, while ultrasound was less efficient, waters of Rio Tinto (Huelva, Spain) to produce lipids and anti-
averaging 67.6% breakage. Steriti et al. [114] treated C. vulgaris with oxidants. This river has an acidic pH (1.7–3.1) and high
838 E.B. D’Alessandro, N.R. Antoniosi Filho / Renewable and Sustainable Energy Reviews 58 (2016) 832–841
concentrations of potentially toxic metals. Authors concluded that The de novo lipid synthesis pathway in microalgae has been
Coccomuxa sp. acidophile can be produced in highly acidic cultures studied lately. Lipids can be produced in three locations: fatty
(pHr2.5), allowing outdoor cultivation with low bacterial acids synthesized in the chloroplast and triacylglyceride (TAG) in
contamination. the endoplasmic reticulum (ER), and thylakoids [44,134]. In the de
Skorupa et al. [120] analyzed the effects of both visible and UV novo lipid synthesis pathway, acetyl-CoA (CoA) is converted into
lights on cellular events of Cyanidioschyzon, a red algae that grow malonyl-CoA by acetyl-CoA-carboxylase (ACCase) combined with
in acidic (pH 0.2–4.0) and hot (38–56 °C) environments. They HCO3-, which is the first step in fatty-acid biosynthesis in various
found that both light wavelengths can be photoinhibitor of organisms. The second step is the conversion of malonyl-CoA into
growth. Thus, wavelength and light intensity must be carefully malonyl-ACP by malonyl-CoA:ACP transacylase (MAT). Posteriorly,
chosen in microalgae cultivation, since they can strongly affect malonyl-ACP is converted into 3-ketoacyl-ACP by 3-ketoacyl-ACP
biomass and lipid productivity. For example, Ho et al. [121] found synthase (KAS), starting the fatty-acid cycle. Finalized by acyl-ACP
that the appropriate light intensity for Scenedesmus obliquus cul- fatty-acid thioesterase (FAT), free fatty acids are exported to the
ture was 420 mmol/m2/s. Conversely, Chen et al. [122] cultivated cytosol. In the chloroplast, acyl-ACP and glycerol 3-phosphate
Chlorella sp. and developed a lumostatic strategy, based on the (G3P) form the diacylglicerids (DAG). Then, the TAGs are expor-
distribution of light detected by image analysis and chlorophyll ted to the cytosol through thylakoids. In the cytosol, the dihy-
content, which increases biomass productivity by 74.3% compared droxyacetone phosphate is converted into G3P by G3P dehy-
to constant light intensity. drogenase (G3PDH). Then, G3P along with acyl-CoA catalyze the
Eutrophic waters with high concentrations of toxic metals are formation of TAG by various enzymes produced by the ER, such as
extreme environments for some microalgae. For example, Park GPAT (glycerol 3-phosphate acyltransferase), LPAAT (lysopho-
et al. [123] cultivated microalgae in eutrophic water from waste- sphatidic acid acyltransferase) LPAT (lysophosphatidylcholine
water treatment effluents. The major difference between acyltransferase), and DAGAT (diacylglycerol acyltransferase).
eutrophic waters and regular culture media is the high con- The enzymes involved in lipid metabolism may be significantly
centration of nitrogen, phosphorus, and toxic metals [124]. Most altered by gene expression levels, because they depend on envir-
nitrogen content in sewage is in form of ammonia, which in high onmental conditions. For example, the expression of the FAD gene
concentrations can inhibit algal growth [125,126]. Sewage waters in H. pluvialis was about six times higher in the low temperature
can still contain metals that inhibit algal growth, such as cadmium, (4 °C) than control (22 °C). This difference in gene expression
zinc, iron, nickel, and copper [127]. Furthermore, such metals may correlated with increasing C18:3ω3 content (1.83 to 2.66 mg/g dry
influence catalysis during biodiesel production or remain in the weight) [46]. Thus, low temperatures can induce the expression of
FAD, raising the content of linolenic acid. Similarly, [116] cultured
final product, producing toxic fumes during biofuel combustion.
Chlamydomonas sp. from Antarctica and found that gene expres-
Some aerial microalgae are also extremophiles, because they
sion of Δ9ACPCiFAD, ω3CiFAD1 and ω3CiFAD2 were more sensi-
can grow on the surface or inside a variety of terrestrial habitats,
tive at 0 °C than 5, 10, and 15 °C, indicating that these genes may
such as rocks, soil, and vascular plants [128,129]. Microalgae are
be involved in a defense mechanism at 0 °C. However, Δ6CiFAD
more exposed to difficult and rapid climate change than aquatic
was more sensitive at 15 °C than at low temperatures, suggesting
ones and produce several carotenoids to adapt to these environ-
that it may be a defense mechanism above 10 °C. The highest fatty
mental conditions (e.g., canthaxanthin, β-carotene, astaxanthin,
acid content in Chlamydomonas sp. at all temperatures was C18:3
and lutein [130,131]). The main stress sources used in carotenoid
(28.2% in mean).
production in aerial microalgae is increasing light intensity and
A way to increase TAG content is to block the metabolic path-
nitrogen deficiency. Aburai et al. [132] found that the color of
ways involved in the production of energetic compounds, such as
cellular content of aerial microalgae changed from green to red
starch [44]. Recently, [135] found a positive correlation between
when cultivated under high light intensity, indicating high pro-
the increase in the enzyme G3PDH and TAG concentration in in
duction of free-ester carotenoids (astaxanthin and adonixanthin),
Phaeodactylum tricornutum growing in nitrogen poor environ-
as well as lutein, canthaxanthin, and β-carotene.
ments. Therefore, to make biodiesel production from microalgae
Thus, culture conditions directly affect not only the levels and
economically feasible it is key to understand the expression pat-
types of pigments, but also fatty acids synthesized by microalgae.
tern of genes involved in lipid metabolism to improve the pro-
Accordingly, pigment production must be reduced to increase fatty
duction and accumulation of FAs and TAGs.
acids production for biodiesel production. Furthermore, lipid
production, especially those with lower unsaturation, should be
stimulated by means of nitrogen depletion [15] and increased 10. Conclusions
water temperature [38].
Microalgae can be used to produce fatty acids and biodiesel. Its
cultivation should consider the following characteristics: (1) avoid
9. Metabolism of fatty acids and acylglycerides using marine microalgae, because they contaminate biodiesel with
sodium and other inorganic compounds; (2) prefer extremophile,
Most of what we known about lipid metabolism in photo- since they are less susceptible to contamination by other micro-
synthetic organisms is based on Arabidopsis [133]. Consequently, organisms (3) microalgae should preferably be able to develop in
Lei et al. [46] used Arabidopsis gene sequences to understand lipid extreme culture media with nutritional disposal, to reduce
metabolism of the microalgae H. pluvialis. It is known that spending with reagents; (4) microalgae must produce large
microalgae accumulate lipids when cell division and growth sta- amounts of biomass in a short term, both at small and large scales;
bilize. For example, Wan et al. [49] found that growth rate (5) species must also withstand stress that induce fatty acid pro-
decreased with increasing lipid accumulation, and Mansour et al. duction, such as temperature or light changes and nutrient
[30] shown that higher lipid accumulation occurred in the linear depletion; (6) cell membrane should have little adherence to the
phase of Gymnodinium sp. cultures, but higher lipid yield took culture system; (7) must cell membrane should allow permeation
place in the stationary phase, due to increased cell density. This of solvents or mechanical action to ease the extraction of fatty acid
relationship is economically important for biodiesel production and other components; (8) should have adequate percentage of
from microalgae. saturated and unsaturated fatty acids; (9) should the microalgae
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