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Contents lists available at ScienceDirect

Journal of Industrial and Engineering Chemistry

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

1
2 Review

3 Sustainable production of liquid biofuels from renewable microalgae

4 biomass
5 Q1 Ok Kyung Lee a, Dong Ho Seong b, Choul Gyun Lee b, Eun Yeol Lee a,*
6 a
Department of Chemical Engineering, Kyung Hee University, Gyeonggi-do 446-701, Republic of Korea
7 b
Marine Bioenergy Research Center, Department of Biological Engineering, Inha University, Incheon 402-751, Korea

A R T I C L E I N F O A B S T R A C T

Article history: Microalgae are important as feedstock in production of liquid biofuels such as biodiesel, bioethanol and
Received 15 February 2015 bio-oil. Biodiesel and bioethanol can be produced from lipids and carbohydrates of microalgae biomass,
Received in revised form 13 April 2015 respectively. Bio-oil and bio-char are prepared using thermochemical treatment of microalgae biomass
Accepted 14 April 2015
or residual biomass after lipid extraction and/or saccharication of cellular carbohydrates. Recent
Available online xxx
advances in biorenery present opportunities to develop sustainable and integrated productions of
various liquid fuels from microalgae biomass in economical way within the next decades. This review
Keywords:
examines the recent progress of microalgae-based liquid biofuel production with regard to
Microalgae
Biodiesel
characteristics and applicability of microalgae as feedstock.
Bioethanol 2015 Published by Elsevier B.V. on behalf of The Korean Society of Industrial and Engineering
Bio-oil Chemistry.
Thermochemical treatment
Q2 Biorenery

8
9 Contents

10 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
11 Biodiesel production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
12 Saccharication and bioethanol fermentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
13 Thermochemical microalgae conversion for bio-oil production . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
14 Biorenery approach for integrated production of liquid fuels from microalgae biomass . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
15 Conclusions and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
16 Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
17 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 000
18

19 Introduction benets. Second, microalgae have high growth rates of 13 28

doublings/day, which is at least ve to ten times higher than those 29
20 Biofuel production from biomass is a promising alternative to of plants. Third, microalgae can use growth nutrients such as 30
21 petroleum-based fuels [1]. Among available biomass sources, nitrogen and phosphorus from waste streams. Additionally, 31
22 microalgae have been frequently considered and investigated as a microalgae can be cultivated from non-productive, non-arable 32
23 third-generation biomass [25]. Microalgae are photoautotrophic lands such as deserts, coasts and offshore marine environments. 33
24 microorganisms that use carbon dioxide, water and sunlight to Large-scale photobioreactors can also be used for microalgae 34
25 grow. Microalgae have several distinct advantages over other cultivation [811]. Thus, large amounts of biomass can be obtained 35
26 biomass sources [6]. First, microalgae can be cultivated using as non-food-based sustainable feedstock from cheap substrates. 36
27 carbon dioxide [7], thus providing greenhouse gas mitigation Production of liquid fuels from microalgae requires many 37
downstream processing steps (Fig. 1). Harvesting is the subsequent 38
step right after cultivation of microalgae and potentially contrib- 39
* Corresponding author. Tel.: +82 31 201 3839; fax: +82 31 204 8114. utes up to 2030% of the total production cost of microalgae 40
E-mail address: eunylee@khu.ac.kr (E.Y. Lee). biomass. Generally, after harvesting step such as centrifugation 41

http://dx.doi.org/10.1016/j.jiec.2015.04.016
1226-086X/ 2015 Published by Elsevier B.V. on behalf of The Korean Society of Industrial and Engineering Chemistry.

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Fig. 1. Downstream processing of biomass harvesting, disruption, and integrated production of biofuels from the major components of microalgae.

42 [12], ltration [13] and occulation [14], microalgae consist of 30% biodiesel is mainly produced from rapeseed and palm oils. 74
43 solid with 70% moisture content [15]. Drying or dewatering of Recently, microalgae are being considered as feedstock for 75
44 microalgae biomass is generally used prior to various conversion biodiesel production [16,18,19]. The lipid content of some 76
45 steps. The energy required for dewatering is known to constitute microalgae species such as Botryococcus braunii exceeds 80% of 77
46 up to 84.9% of total energy consumption [16]. Extraction of lipid is the dry weight [20]. Chlorella and Dunaliella are known to have lipid 78
47 required for biodiesel production. contents as great as 50% of the dry weight. Thus, considerable 79
48 The main components of microalgae for liquid biofuel produc- amounts of lipid for biodiesel production can be obtained from 80
49 tion are lipids, carbohydrates and others such as proteins. Various large-scale cultivation of microalgae. As indicated in some reports, 81
50 conversion methods including transesterication, fermentation, the yield of microalgae lipid per hectare is around 58,700 l/ha, 82
51 pyrolysis, liquefaction and anaerobic digestion are used for almost ten times greater than those from palm crops [21]. 83
52 production of biodiesel, bioethanol, bio-oil and methane (Fig. 2). Biodiesel can be produced using a two-step method of oil 84
53 The lipid is extracted using organic solvent and then transesteried extraction-transesterication or one-step transesterication (so 85
54 to biodiesel in the presence of base or acid catalysts. Bioethanol is called in site or direct transesterication). The common method 86
55 produced from microalgae fermentation, while methane can be for extracting oil from microalgae is a solvent extraction using 87
56 produced through anaerobic digestion. Thermochemical processes hexane, ethanol, methanol and methanolchloroform mixture 88
57 such as hydrothermal liquefaction and pyrolysis can be applied for (2:1 v/v) [22]. The extraction efciency of microalgae oil using 89
58 bio-oil production. Currently, no liquid biofuel production from n-hexane is rather low, although it is widely used for oil extraction 90
59 microalgae is commercially implemented because liquid fuel from seed crops [23]. Ultrasonic-assisted extraction, microwave- 91
60 production only is not economically feasible. Thus, the whole assisted extraction and supercritical uid extraction can be an 92
61 components of microalgae biomass should be used to produce alternative to organic solvent extraction. After lipid extraction, 93
62 multiple products including various liquid biofuels for economi- alkali, acid catalysts and lipase are used for transesterication. One 94
63 cally viable option [17]. In this review paper, recent progress on the important thing is microalgae oil generally contains a certain 95
64 production of liquid biofuels, including biodiesel, bioethanol and amount of free fatty acid. Thus, an acid-catalyzed conversion can 96
65 bio-oil, are addressed, and some concluding remarks are also be an efcient method with high conversion, though its reaction 97
66 discussed for a successful zero-waste microalgae biorenery. rate is approximately 4000 times slower than base-catalyzed 98
processes [24]. 99
67 Biodiesel production Recent advances in biodiesel production from microalgae are 100
summarized in Table 1. In two-step transesterication, microalgae 101
68 Biodiesel is a mixture of fatty acid methyl esters (FAMEs) lipids were extracted using different solvents and extraction 102
69 produced by transesterication of triglycerides with alkyl accep- methods from various microalgae strains [2532]. The extracted 103
70 tors such as methanol in the presence of a catalyst, usually NaOH, lipids were transesteried to FAMEs using H2SO4, HCl and lipase. 104
71 KOH or lipase. Since methanolysis of triglyceride is an equilibrium An acid catalyst was used because microalgae biomass generally 105
72 reaction, a high excess of methanol, generally six moles of contains large amounts of free fatty acids that cause the formation 106
73 methanol per mole of triglyceride, is applied [6]. At present, of soaps in the presence of alkali catalysts [25]. Lam et al. [26] 107

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Fig. 2. Various biochemical and thermochemical approaches for production of liquid biofuels from microalgae.

Table 1
Biodiesel production from various microalgae.

Method Microalgae species (dry or Biodiesel production process Catalyst Lipid Biodiesel Reference
wet) content (%)a conversion (%)b

Two-step Nannochloropsis gaditana Stirring extraction Transesterication solvent HCl 13.1 41.8 [25]
(dry biomass) Chloroformmethanol Chloroformmethanol
Phaeodactylum Tricornutum Ultrasonic assisted Transesterication solvent HCl 24.5 37.5
(dry biomass) extraction Chloroformmethanol
Chloroformmethanol
Chlorella vulgaris (dry Stirring extraction Transesterication solvent H2SO4 95.0 [26]
biomass) Chloroformmethanol Methanol-THF
Chlorella protothecoides Soxhlet extraction Transesterication solvent lipase 4448.7 98.1 [27]
(dry biomass) Hexane Hexane (Candidia sp.)
Chlorella vulgaris (dry Soxhlet extraction Transesterication solvent IL lipase 40.7 90.7 [28]
biomass) Hexane ([BMIm][PF6]) (P. expansum)
Chlorella vulgaris ESP-31 Ultrasonic assisted Transesterication solvent lipase 63.2 72.1 [29]
(dry biomass) extraction Hexane (Burkholderia sp.)
Chloroformmethanol
Chlorella sp. KR-1 (dry Stirring extraction Transesterication solvent lipase 38.9 75.5 [30]
biomass) DMCmethanol DMC (Novozyme 435)
Tribonema minus (wet Subcritical ethanol Transesterication solvent H2SO4 20.2 96.5 [31]
biomass) extraction Methanol
Nannochloropsis sp. Microwave assisted Transesterication solvent NaOH 38.3 86.4 [32]
(80% moisture) extraction Hexane Methanolhexane
methanol
One step Chlorella sp. KR-1 (dry DMC based lipase (Novozyme 435) 40.9 89.7 [33]
biomass) transesterication
Chlorella protothecoides Non-catalytic 90.8 (87.8) [34]
(dry biomass) transesterication in
supercritical methanol
(ethanol)
Tetraselmis suecica Microwave assisted H2SO4 23.0 78.0 [35]
(dry biomass) Chlorella transesterication
sorokiniana (dry biomass)
23.5 77.0
Nannochloropsis oceanica In situ H2SO4 19.1 90.6 [36]
(65% moisture) transesterication in
water bath
Chlorella pyrenoidosa Microwave assisted H2SO4 19.0 55.2 [37]
(80% moisture) transesterication
a
Lipid content = (total lipid/total biomass)  100.
b
Biodiesel conversion = (weight of FAMEs/total lipid)  100.

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108 reported that more than 95% biodiesel yield from highly viscous Saccharication and bioethanol fermentation 174
109 lipids (extracted from Chlorella vulgaris) was achieved by
110 introducing tetrahydrofuran as co-solvent. The lipids extracted Microalgae contain large amounts of various carbohydrates 175
111 from Chlorella protothecoides using hexane were converted to such as starch, cellulose and hemicellulose, depending on the 176
112 FAMEs by lipase, with biodiesel conversion as high as 98.15% [27], species [40]. Microalgae such as Chlorella, Dunaliella, Spirulina and 177
113 because lipase generally showed greater activity toward hexane- others are known to contain carbohydrates in the form of starch 178
114 extracted lipids consisting mainly of triglycerides with very small and glycogen in amounts as great as 50% of the dry weight [41,42]. 179
115 amounts of free fatty acids. Ionic liquid was successfully used for Carbohydrates can be used for bioethanol fermentation [43]. 180
116 lipid extraction from C. vulgaris, with a 40.7% yield based on dry cell Microalgae biomass has several advantages over other biomasses 181
117 weight [28]. Lee et al. [30] reported a method for the highly such as lignocellulose. The most advantageous feature of micro- 182
118 efcient lipid extraction and lipase-catalyzed transesterication of algae is that they contain little or no lignin [42], which mitigates 183
119 lipids from Chlorella sp. KR-1 using dimethyl carbonate (DMC). cost-intensive biomass pretreatment. Additionally, carbon dioxide 184
120 With a mixture of DMC and methanol at a ratio of 7:3, total lipids produced as a byproduct of bioethanol fermentation can be used to 185
121 were almost completely extracted under simple stirring at mild cultivate microalgae. 186
122 conditions of 50 8C. Biodiesel conversion of 75.5% was obtained Saccharication is one of the most critical steps for economi- 187
123 using lipase. cally viable bioethanol production. Acid hydrolysis, the most 188
124 Recently, direct lipids extractions from wet microalgae were commonly applied method, is conducted at 121 8C in the presence 189
125 conducted to minimize the energy required for microalgae of H2SO4 and HCl (15%) [44,45]. Acid hydrolysis provides high 190
126 biomass drying. Lipids were extracted from wet paste of Tribonema sugar yields up to 100% for short time. However, acid hydrolysate 191
127 minus using subcritical ethanol extraction, and then converted to needs to be neutralized before fermentation. On the contrary, 192
128 biodiesel through acid-alkaline transesterication [31]. Washidin enzymatic saccharication is conducted at mild condition and can 193
129 et al. [32] reported that 38.3% (w/w) of lipids in wet microalgae was be liked to fermentation, simultaneous saccharication and 194
130 extracted by microwave irradiation. The subsequent biodiesel fermentation (SSF). Each species of microalgae has different 195
131 conversion using microwave irradiation was 86.4% based on the polysaccharide composition, thus suitable enzymes such as 196
132 extracted lipid. cellulase, glucanase, xylanase and etc. should be employed. 197
133 In case of one step transesterication, Jo et al. [33] developed a Various microalgae have been used for saccharication and 198
134 DMC-based direct transesterication for Chlorella sp. KR-1 biomass bioethanol fermentation (Table 2) [4553]. One advantage in a 199
135 in the presence of lipase. A 367.31 mg of biodiesel was obtained saccharication of microalgae biomass is that intensive pretreat- 200
136 from 1 g of Chlorella sp. KR-1 containing 40% (w/w) lipid. ment is not required. Various chemical saccharications have 201
137 Interestingly, the biodiesel yield of direct transesterication was been employed for microalgae. Harun et al. [46] used alkali 202
138 increased 1.25-fold, compare with two step transesterication saccharication for bioethanol production from Chlorococcum 203
139 (293.82 mg of biodiesel) [30,33]. To enhance the biodiesel infusionum. The highest glucose yield was 0.35 g glucose/g 204
140 conversion, supercritical uid was applied. Nan et al. [34] reported microalgae using 0.75% (w/v) of NaOH and 120 8C for 30 min, 205
141 a production of biodiesel with 90.8% yield through non-catalytic and the maximum bioethanol yield was 0.26 g ethanol/g 206
142 transesterication of microalgae lipid in supercritical methanol. microalgae. Harun and Danquah [47] investigated the inuence 207
143 For direct transesterication of wet microalgae, Im et al. [36] of acid saccharication on Chlorococcum humicola for bioethanol 208
144 reported a direct transesterication of wet Nannochloropsis production. A 7.2 g/L bioethanol was obtained after 15 g/L 209
145 oceanica (65% moisture) with 91.1% biodiesel conversion in the biomass was treated using 1% (v/v) of sulfuric acid at 140 8C for 210
146 presence of 0.3 g H2SO4, 2 ml chloroform, and 1 ml methanol at 30 min. A 40 g/L Scenedesmus obliquus CNW-N containing 51.8% 211
147 95 8C for 90 min. Microwave has been employed for an one-step (w/w) carbohydrate was hydrolyzed using 2% (v/v) H2SO4. The 212
148 transesterication of Chlorella pyrenoidosa (80% moisture) in the resulting broth was fermented by Zymomonas mobilis, and an 213
149 presence of chloroform, methanol and sulfuric acid as the catalyst. 8.6 g/L bioethanol was produced at a 21.3% yield (g ethanol/g 214
150 The biodiesel conversion with the aid of microwave was 1.3-fold biomass) [48]. A 14.6 g/L bioethanol was produced from 50 g/L 215
151 higher than that of conventional two-step heating method [37]. biomass of Chlamydomonas reinhardtii UTEX 90 by Saccharomyces 216
152 The fuel properties of microalgae biomass-based biodiesel such cerevisiae with 29.2% yield after the biomass was saccharied 217
153 as density, viscosity, ash point, cold lter plugging point, heating using 3% (v/v) H2SO4 at 110 8C for 30 min [49]. Lee et al. [50] 218
154 value and solidifying point have been well characterized [38]. The hydrolyzed C. vulgaris using 5% (v/v) H2SO4 at 110 8C for 120 min. 219
155 heating value of microalgae-based biodiesel was approximately The maximum ethanol yield of 0.4 g ethanol/g biomass was 220
156 3141 MJ/kg, a little bit low compared to that of petroleum-based achieved using ethanolic E. coli SJL2526. Carbohydrate-rich C. 221
157 diesel (46 MJ/kg). One interesting characteristics of microalgae- vulgaris FSP-E containing 51% carbohydrate per dry weight was 222
158 based biodiesel is its lower cold lter plugging point of 11 8C, used as feedstock for bioethanol fermentation [51]. Dilute acidic 223
159 more suitable fuel for aviation than petroleum-based diesel. On the (1% H2SO4) and enzymatic saccharication of C. vulgaris FSP-E 224
160 other hand, viscosity of microalgae biodiesel is high, approximate- biomass produced glucose yields of 93.6 and 90.4%, respectively. A 225
161 ly 1.22.6 times more than that of petroleum-based diesel. The 11.7 g/L bioethanol with 87.6% theoretical yield was obtained 226
162 composition of biodiesel from microalgae depends on the species, from the acidic hydrolysate of a 50 g/L biomass using a separate 227
163 which also strongly affects fuel properties like viscosity. Thus, hydrolysis and fermentation (SHF) process. The enzymatic 228
164 selection and genetic improvement of microalgae species contain- hydrolysate was converted into bioethanol by both SHF and SSF 229
165 ing an appropriate fatty acid composition needs to be conducted to processes, producing 79.9 and 92.3% theoretical yields, respec- 230
166 lower the fatty acid composition for lower viscosity. Although the tively. Choi et al. [52] used enzyme saccharication using two 231
167 fuel properties of biodiesel is similar to petro-diesel, biodiesel different commercial enzymes (including amylase from Bacillus 232
168 production from microalgae is still too expensive to be commer- licheniformis and glucoamylase from Aspergillus niger) for 233
169 cialized based on a commercial feasibility study. Davis et al. bioethanol production from C. reinhardtii UTEX 90. A sugar 234
170 reported that the production of microalgae biodiesel could cost conversion of 0.5 g sugar/g microalgae was obtained and 0.23 g 235
171 from $9.84 (open pond) to $20.53 (photobioreactor) per gallon, ethanol was produced from 1.0 g microalgae by SHF method. 236
172 much higher than $2.60 per gallon for petroleum diesel production Likewise, many investigations have been successfully conducted 237
173 [39]. in order to commercialize bioethanol production from microalgae. 238

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Table 2
Saccharication and bioethanol fermentation of microalgae biomass.

Microalgae Hydrolysis Method Biomass Fermentation Carbohydrates Yeast Ethanol Ethanol yield Reference
species concentration type (%) [glucose strain (g/L) (%, g ethanol/g
(g/L) (g/L)] biomass)

Chlorococcum Chemical 50 SHF 43.9 [35] S. cerevisiae N.D 26 [46]

infusionum 0.75%(w/v) NaOH
Chlorococcum Chemical 15 SHF 32.5 S. cerevisiae 7.2 52 [47]
humicola 1% (v/v) H2SO4
Scenedesmus Chemical 40 SHF 51.8 Z. mobilis 8.6 21 [48]
obliquus 2% (v/v) H2SO4
CNW-N
Chlamydomonas Chemical 50 SHF 60 [28.5] S. cerevisiae 14.6 29 [49]
reinhardtii 3% (v/v) H2SO4
UTEX 90
Chlorella vulgaris Chemical N.D SHF N.D E.coli N.D 40 [50]
5% (v/v) H2SO4
Chlorella vulgaris Chemical 50 SHF 4748 [23.6] Z. mobilis 11.7 23 [51]
FSP-E 1% (v/v) H2SO4
Enzymatic 20 SHF 43.546.8 [7.78] Z. mobilis 3.6 18
Cellulase + Amylases
Enzymatic 20 SSF N.D Z. mobilis 4.3 21
Cellulase + Amylases
Chlamydomonas Enzymatic 50 SHF 59.7 [25.21] S. cerevisiae 11.7 23 [52]
reinhardtii Termamyl 120L
UTEX 90 +AMG 300L
Dunaliella tertiolecta Enzymatic AMG 50 SHF 51.9 [42.0] S. cerevisiae 7.2 14 [45]
(lipid-extracted 300L
microalgae)
Chlorococcum sp. Physical 10 SHF N.D S. bayanus 3.8 38 [53]
(lipid-extracted Supercritical CO2
microalgae)

239 However, the commercial production of bioethanol from microalgae because its feedstock is wet biomass itself instead of dry matters. 250
240 is still in the research stages, similar to biodiesel production from The drying step of biomass is one of the most energy consuming 251
241 microalgae. processes in fuels production process from microalgae. The main 252
product of the HTL is bio-oil namely biocrude, a liquid crude fuel 253
242 Thermochemical microalgae conversion for bio-oil production composed of many hydrocarbon compounds. Bio-oil is organic 254
solvent extractable black liquid oil with a high heating value and 255
243 Representative two thermochemical processes for bio-oil high viscosity. The reported high bio-oil production yield of 256
244 production using microalgae biomass are hydrothermal liquefac- microalgae HTL is 5060% based on dry biomass [54]. The microalgae 257
245 tion (HTL) and pyrolysis process (Fig. 3). The HTL is conducted at bio-oil is typically composed of 7075% carbon, 1016% oxygen, 258
246 subcritical conditions in water, for obtaining liquid fuels from 46% nitrogen, and 0.51% sulfur. It has higher heating value (HHV) 259
247 biomass. During the HTL reaction, water is acted as both a reaction of 33.439.9 MJ/kg. The major components of bio-oil are aromatic 260
248 solvent and reagent for hydrolysis of biomass. The HTL have hydrocarbons, long chain fatty acids, nitrogen heterocyclic com- 261
249 relatively good efciency of energy recovery on energy investment pounds, alcohols, organic acid, aldehydes and etc.[55]. The HHV of 262

Fig. 3. Production of bio-oil from thermochemical treatment of microalgae.

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263 microalgae bio-oil is relatively high because of low content of product. In contrast, slow pyrolysis produces more bio-char [62]. 304
264 oxygen. The characteristics of microalgae bio-oil as a fuel and One disadvantage of the pyrolysis process is the high capital cost of 305
265 production yield are closely affected by the microalgae composition separation equipment for various fractions. Another signicant 306
266 as a feedstock, reaction temperature, time, catalyst and extraction hurdle in using pyrolysis for microalgae is the high moisture 307
267 method of bio-oil from reactant. When the same microalgae biomass content of biomass feedstock, which requires dewatering to 308
268 is used as a HTL feedstock, the temperature is the most critical control moisture content of the bio-oil. Development of an 309
269 factor among the inuencing conditions for production of microalgae inexpensive dewatering or extraction process is a key factor for 310
270 bio-oil. cost-competitive pyrolysis. Reports on pyrolysis of microalgae for 311
271 The typical reaction temperature for microalgae HTL is 300 bio-oil production are less available compared to those for 312
272 374 8C, a subcritical temperature, and as the reaction temperature lignocellulosic biomass. Pyrolysis converts triglycerides to fatty 313
273 is elevated within that subcritical water temperature, the bio-oil acid alkyl esters as bio-oil [63]. Microalgae carbohydrate and 314
274 yield and nitrogen content in bio-oil are increased and the oxygen protein components as well as fat are converted to bio-oil by 315
275 content is decreased [56,57]. When the reaction temperature pyrolysis [64]. Bio-oil was obtained at a 57.9% yield from fast 316
276 exceeds the critical point, the bio-oil is produced less and gas takes pyrolysis of C. protothecoides at a heating value of 41 MJ/kg [65]. 317
277 place more. The microalgae bio-oil produced through HTL is more Bio-oil from HTL and pyrolysis can be used as drop-in fuel in 318
278 suitable liquid fuel than that produced by pyrolysis method industrial boilers for heat production [66,67]. To use bio-oil 319
279 because not only it contains a less oxygen and water in bio-oil but directly as transportation fuel, catalytic hydrogenation can be 320
280 also it has higher storage stability than that of pyrolysis oil [58]. applied to upgrade the bio-oil [68]. Lower oxygen content and a 321
281 Although bio-oil from HTL contains less nitrogen and sulfur than higher heating value were obtained using a catalyst [69,70]. 322
282 the feed microalgae biomass, nitrogen and sulfur contents in bio- Carbon yield of aromatic hydrocarbons increased from 0.9 to 25.8% 323
283 oil are still in a high level. Bio-oil contains a large quantity of in the pyrolysis of C. vulgaris using zeolite HZSM-5 [71]. When 324
284 double bonds, so upgrading needs to be conducted for practical use ZSM-5 catalyst was used with an increased catalyst to biomass 325
285 as liquid fuel. Hydrotreatment, a hydrogen addition process, is ratio, oxygen content in the bio-oil decreased from 30.1 to 326
286 being used for upgrading bio-oil. Generally, although HTL and 19.5 wt%, and the heating value of the bio-oil improved to 24.4 327
287 pyrolysis have their own advantages, HTL is expected to be more 32.2 MJ/kg [72]. 328
288 cost-effective for wet microalgae biomass due to the omitting of
289 energy-intensive drying process in the thermal conversion. Biorenery approach for integrated production of liquid fuels 329
290 Pyrolysis is a thermochemical conversion of biomass to bio-oil, from microalgae biomass 330
291 bio-char and gas at medium to high temperatures (generally 450
292 500 8C) with short residence time (order of seconds to minutes) in Biorenery is a process of integrated utilization of every 331
293 the absence of oxygen [59,60]. Contrary to HTL, biomass drying is component of the biomass in order to enhance the economics of 332
294 necessary (Fig. 3). Pyrolysis of microalgae can produce a wide the process [73,74]. In a biorenery process, major components of 333
295 range of products including liquid oil, solid char and gas, microalgae are subsequently used to produce biofuels, chemicals 334
296 depending on the reaction parameters such as heating rate, and feeds [75,76]. Some biologically active materials such as 335
297 residence time, and pyrolysis temperature. In fast pyrolysis, astaxanthin or tocopherol that are used as additives for nutra- 336
298 according to the vapor residence time, nely ground biomass is ceuticals and cosmetics are produced from microalgae. However, 337
299 treated for short residence times and fast heating rates at 350 the amount of those biomaterials to meet market demand does not 338
300 500 8C [61]. One major advantage of pyrolysis over other require large scale production using biorenery process for 339
301 conversion processes is fast biomass conversion on the order of biofuels and chemicals. In case of feed production, there might 340
302 seconds to minutes. Fast pyrolysis with short residence times and be some problem in safety and approval when animal feed 341
303 fast heating rates tends to favor formation of bio-oil as the main production is integrated with the production of biofuels and 342

Fig. 4. Sequential production of biodiesel, bioethanol and bio-oil from microalgae biomass.

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343 chemicals. Thus, an integrated production of various liquid microalgae biomass, extraction and separation of cell components 407
344 biofuels in large scale based on biorenery concept needs to be and conversion of the components to biofuels need to be 408
345 developed from the industrial point of view. integrated. Less energy-intensive pretreatment processes such 409
346 All cellular components in microalgae biomass should be as cost-effective harvesting and dewatering prior to extraction and 410
347 utilized to produce valuable liquid fuels in order to reduce overall conversion need to be developed. A cost-effective extraction 411
348 production costs. Each cellular component of microalgae has own method is still required even though microalgae biomass is lignin- 412
349 advantage for different liquid fuels. Lipid is suitable for production free. Recruit of new methods for extraction of oil directly from wet 413
350 of biodiesel and bio-oil, whereas polysaccharide is appropriate for microalgae biomass without energy-intensive processing are 414
351 bioethanol. Recently, sequential production of biodiesel, bioetha- important. Along with oil for biofuel, other substances such as 415
352 nol and bio-oil were successfully conducted for Dunaliella carbohydrates and proteins should be used to improve biorenery 416
353 tertiolecta biomass (Fig. 4) [45,77]. The lipids of D. tertiolecta were process economics. Fully integrated production of biodiesel, 417
354 extracted using a chloroform and methanol mixture and then bioethanol, bio-oil and bio-char can be a key solution for zero- 418
355 converted to biodiesel. The residual biomass after lipid extraction waste biorenery of microalgae. In the near future, biorenery for 419
356 has been successfully used as raw material for saccharication and biofuel production from microalgae is expected to play an 420
357 subsequent bioethanol fermentation due to the large amount of important role in the bio-based economy. 421
358 polysaccharides present in the residual biomass. The residual
359 biomass was composed of 51.9 wt% carbohydrate, 35.0 wt% Acknowledgements 422
360 protein, 13.1 wt% ash and no trace of lipids. The residual biomass
361 was saccharied using an AMG 300 L enzyme with a saccharica- This research was supported by Basic Science Research Program 423
362 tion yield of 80.9% (w/w) based on the total amount of through the National Research Foundation of Korea (NRF) funded 424
363 carbohydrates [45]. Without any pretreatment, bioethanol was by the Ministry of Science, ICT and Future Planning (NRF- Q3425
364 directly produced from the enzymatic saccharication products 2013R1A2A2A01068863). This work was supported by a grant 426
365 with a fermentation yield of 0.14 g ethanol/g residual biomass, from the Marine Biotechnology Program funded by the Ministry of 427
366 which was 82.0% of the theoretical fermentation yield. For Oceans and Fisheries of the Korean Government. Q4428
367 complete utilization of D. tertiolecta biomass, the residual biomass
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