Comparative analyses of three Chlorella species in response to light and sugar reveal distinctive lipid accumulation patterns in the microalga C. sorokiniana

PLoS ONE

Volumn 9, Issue 4, 2014, Pages

  Rosenberg, Julian N ^a,b   Kobayashi, Naoko ^c   Barnes, Austin ^c   Noel, Eric A ^c   Betenbaugh, Michael J ^a   Oyler, George A ^a,b,c  

^a Whiting School of Engineering   (United States)

^b SYNAPTIC RESEARCH LLC   (United States)

^c UNIVERSITY OF NEBRASKA LINCOLN   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

GLUCOSE; LIPID; SUGAR; TRIACYLGLYCEROL; BIOFUEL; CARBOHYDRATE;

ALGAL GROWTH; ARTICLE; BIOMASS; CHLORELLA; CHLORELLA PROTOTHECOIDES; CHLORELLA SOROKINIANA; CHLORELLA VULGARIS; COMPARATIVE STUDY; CONTROLLED STUDY; DRY WEIGHT; GROWTH RATE; LIGHT; LIPID COMPOSITION; LIPID STORAGE; MOLECULAR PHYLOGENY; NONHUMAN; SPECIES CULTIVATION; BIODIVERSITY; BIOSYNTHESIS; CLASSIFICATION; DRUG EFFECTS; GROWTH, DEVELOPMENT AND AGING; HETEROTROPHY; MASS FRAGMENTOGRAPHY; METABOLISM; PHYLOGENY; RADIATION RESPONSE;

BIODIVERSITY; BIOFUELS; BIOMASS; CARBOHYDRATES; CHLORELLA; GAS CHROMATOGRAPHY-MASS SPECTROMETRY; HETEROTROPHIC PROCESSES; LIGHT; LIPIDS; PHYLOGENY;

EID: 84899080970     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0092460     Document Type: Article

References (117)

1. Blankenship RE, Tiede DM, Barber J, Brudvig GW, Fleming G, et al. (2011) Comparing photosynthetic and photovoltaic efficiencies and recognizing the potential for improvement. Science 332: 805-809.
2. Zhu X-G, Long SP, Ort DR (2008) What is the maximum efficiency with which photosynthesis can convert solar energy into biomass? Curr Opin Biotech 19: 153-159.
3. Bumbak F, Cook S, Zachleder V, Hauser S, Kovar K (2011) Best practices in heterotrophic high-cell-density microalgal processes: achievements, potential and possible limitations. Appl Microbiol Biot 91: 31-46.
4. Senanayake SPJN, Ahmed N, Fichtali J (2010) Chapter 37. Nutraceuticals and Bioactives from Marine Algae. In: C . Alasavar, F . Shahidi, K . Miyashita and U . Wanasundara, editors. Handbook of Seafood Quality, Safety and Health Applications. Oxford, UK: Blackwell Publishing Ltd. pp. 455-462.
5. Patnaik S, Samocha TM, Davis DA, Bullis RA, Browdy CL (2006) The use of HUFA-rich algal meals in diets for Litopenaeus vannamei. Aquacult Nutr 12: 395-401. (Pubitemid 44337502)
6. Davis R, Fishman D, Frank ED, Wigmosta MS, Aden A, et al. (2012) Renewable Diesel from Algal Lipids: An Integrated Baseline for Cost, Emissions, and Resource Potential from a Harmonized Model. Technical Report Numbers: ANL/ESD/12-4; NREL/TP-5100-55431; PNNL-21437. Argonne, IL: Argonne National Laboratory; Golden, CO: National Renewable Energy Laboratory; Richland, WA: Pacific Northwest National Laboratory.
7. Schenk PM, Thomas-Hall SR, Stephens E, Marx UC, Mussgnug JH, et al. (2008) Second generation biofuels: high-efficiency microalgae for biodiesel production. Bioenerg Res 1: 20-43.
8. Wu Q, Miao X (2006) Biodiesel production from heterotrophic microalgal oil. Bioresource Technol 97: 841-846. (Pubitemid 43042411)
9. Li X, Xu H, Wu Q (2007) Large-scale biodiesel production from microalga Chlorella protothecoides through heterotrophic cultivation in bioreactors. Biotechnol Bioeng 98: 764-771. (Pubitemid 350032681)
10. Harwood JL, Guschina IA (2009) The versatility of algae and their lipid metabolism. Biochimie 91: 679-684.
11. Niederholtmeyer H, Wolfstadter BT, Savage DF, Silver PA, Way JC (2010) Engineering cyanobacteria to synthesize and export hydrophilic products. Appl Environ Microb 76: 3462-3466.
12. National Research Council (2012) Sustainable Development of Algal Biofuels in the United States. In: L.G . Coplin, editor. Washington DC: The National Academies Press. Available: http://www.nap.edu/catalog.php?record-id= 13437. Accessed 2012 Oct 25.
13. Vasudevan V, Stratton RW, Pearlson MN, Jersey GR, Beyene AG, et al. (2012) Environmental performance of algal biofuel technology options. Environ Sci Technol 46: 2451-2459.
14. Converti A, Casazza AA, Ortiz EY, Perego P, Borghi MD (2009) Effect of temperature and nitrogen concentration on the growth and lipid content of Nannochloropsis oculata and Chlorella vulgaris for biodiesel production. Chem Eng Process 48: 1146-1151.
15. Msanne J, Xu D, Konda AR, Casas-Mollano JA, Awada T, et al. (2011) Metabolic and gene expression changes triggered by nitrogen deprivation in the photoautotrophically grown microalgae Chlamydomonas reinhardtii and Coccomyxa sp. C-169. Phytochemistry 75: 50-59.
16. Sheehan J, Dunahay T, Benemann J, Roessler P (1998) A Look Back at the U.S. Department of Energy's Aquatic Species Program: Biodiesel from Microalgae. Report Number: TP-580-24190. Golden, Colorado: National Renewable Energy Lab.
17. Stephenson AL, Dennis JS, Howe CJ, Scott SA, Smith AG (2010) Influence of nitrogen-limitation regime on the production by Chlorella vulgaris of lipids for biodiesel feedstocks. Biofuels 1: 47-58.
18. Spoehr HA, Milner HW (1949) The chemical composition of Chlorella; effect of environmental conditions. Plant Physiol 24: 120-149.
19. Perez-Garcia O, Escalante FME, de-Bashan LE, Bashan Y (2011) Heterotrophic cultures of microalgae: metabolism and potential products. Water Res 45: 11-36.
20. Samejima H, Meyers J (1958) On the heterotrophic growth of Chlorella pyrenoidosa . J Gen Microbiol 18: 107-117.
21. Xiong W, Liu L, Wu C, Yang C, Wu Q (2010) 13C-tracer and gas chromatography-mass spectrometry analyses reveal metabolic flux distribution in the oleaginous microalga Chlorella protothecoides. Plant Physiol 154: 1001-1011.
22. Fan J, Huang J, Li Y, Han F, Wang J, et al. (2012) Sequential heterotrophy-dilution-photoinduction cultivation for efficient microalgal biomass and lipid production. Bioresource Technol 112: 206-211.
23. Zheng Y, Chi Z, Lucker B, Chen S (2012) Two-stage heterotrophic and phototrophic culture strategy for algal biomass and lipid production. Bioresource Technol 103: 484-488.
24. Lee Y-K (2006) Ch. 7 Algal Nutrition: Heterotrophic Carbon Nutrition. In: A . Richmond, editor. Handbook of Microalgal Culture: Biotechnology and Applied Phycology. Oxford, UK: Backwell Science Ltd. pp. 116-124.
25. Wan M-X, Wang R-M, Xia J-L, Rosenberg JN, Nie Z-Y, et al. (2012) Physiological evaluation of a new Chlorella sorokiniana isolate for its biomass production and lipid accumulation in photoautotrophic and heterotrophic cultures. Biotechnol Bioeng 109: 1958-1964.
26. Kiron V, Phromkunthong W, Huntley M, Archibald I, Scheemaker GD (2012) Marine microalgae from biorefinery as a potential feed protein source for Atlantic salmon, common carp and whiteleg shrimp. Aquacult Nutr 18: 521-531.
27. Tredici MR, Biondi N, Ponis E, Rodolfi L (2009) Chapter 20: Advances in microalgal culture for aquaculture feed and other uses. In: G . Burnell, G . Allan, editors. New technologies in aquaculture production efficiency, quality and environmental management. Boca Raton, FL: CRC Press. pp. 610-676.
28. Gouveia L, Batista AP, Sousa, Raymundo A, Bandarra NM (2008) Microalgae in novel food products. In: N . Konstantinos, P.P . Papadopoulos, editors. Food Chemistry Research Developments. New York, NY: Nova Science Publishers.
29. Warburg O, Negelein E (1920) Uber die Reduktion der Salpetersaure in griinen Zellen. Biochem Z 110: 66-115.
30. Kessler E (1976) Comparative physiology, biochemistry, and the taxonomy of Chlorella (Chlorophyceae). Plant Syst Evol 125: 129-138.
31. Burlew JS (1953) Algal Culture from Laboratory to Pilot Plant. Washington DC: Carnegie Institution of Washington.
32. Blanc G, Duncan G, Agarkova I, Borodovsky M, Gurnon J, et al. (2010) The Chlorella variabilis NC64A genome reveals adaptation to photosymbiosis, coevolution with viruses, and cryptic sex. Plant Cell 22: 2943-2955.
33. McAuley PJ (1986) Glucose uptake by symbiotic Chlorella in the green-hydra symbiosis. Planta 168: 523-529.
34. Muscatine L, Karakashian SJ, Karakashian MW (1967) Soluble extracellular products of algae symbiotic with a ciliate, a sponge, and a mutant hydra. Comp Biochem Physiol 20: 1-12.
35. Ziesenisz E, Reisser W, Wiessner W (1981) Evidence of de novo synthesis of maltose excreted by the endosymbiotic Chlorella from Paramecium bursaria. Planta 153: 481-485.
36. Dunigan DD, Cerny RL, Bauman AT, Roach JC, Lane LC, et al. (2012) Paramecium bursaria Chlorella virus 1 proteome reveals novel architectural and regulatory features of a giant virus. J Virol 86: 8821-8834.
37. Van Etten JL (2003) Unusual life style of Chlorella viruses. Annu Rev Genet 37: 153-195.
38. Yamada T, Onimatsu H, Van Etten JL (2006) Chlorella viruses. Adv Virus Res 66: 293-336. (Pubitemid 44107835)
39. Shi X, Wu Z, Chen F (2006) Kinetic modeling of lutein production by heterotrophic Chlorella at various pH and temperatures. Mol Nutr Food Res 50: 763-768. (Pubitemid 44348626)
40. Shi X-M, Zhang X-W, Chen F (2000) Heterotrophic production of biomass and lutein by Chlorella protothecoides on various nitrogen sources. Enzyme Microb Technol 27: 312-318. (Pubitemid 30427206)
41. Ip P-F, Chen F (2005) Production of astaxanthin by the green microalga Chlorella zofingiensis in the dark. Process Biochem 40: 733-738. (Pubitemid 39437875)
42. Sun N, Wang Y, Li Y-T, Huang J-C, Chen F (2008) Sugar-based growth, astaxanthin accumulation and carotenogenic transcription of heterotrophic Chlorella zofingiensis (Chlorophyta). Process Biochem 43: 1288-1292.
43. Feng P, Deng Z, Fan L, Hu Z (2012) Lipid accumulation and growth characteristics of Chlorella zofingiensis under different nitrate and phosphate concentrations. J Biosci Bioeng 114: 405-410.
44. Matsukawa R, Hotta M, Masuda Y, Chihara M, Karube I (2000) Antioxidants from carbon dioxide fixing Chlorella sorokiniana. J Appl Phycol 12: 263-267.
45. Guarnieri MT, Nag A, Smolinski SL, Darzins A, Seibert M, et al. (2011) Examination of triacylglycerol biosynthetic pathways via de novo transcriptomic and proteomic analyses in an unsequenced microalga. PLoS ONE (6:10): e25851.
46. Wan M, Faruq J, Rosenberg JN, Xia J, Oyler GA, et al. (2012) Achieving high throughput sequencing of cDNA library utilizing an alternative protocol for the bench top next-generation sequencing system. J Microbiol Meth 92: 122-126.
47. Fan J, Cui Y, Huang J, Wang W, Yin W, et al. (2012) Suppression subtractive hybridization reveals transcript profiling of Chlorella under heterotrophy to photoautotrophy transition. PLoS ONE 7: e50414.
48. Takeda H (1988) Classification of Chlorella strains by cell wall sugar composition. Phytochemistry 27: 3823-3826.
49. Banskota AH, Stefanova R, Gallant P, Osbornw JA, Melanson R, et al. (2013) Nitric oxide inhibitory activity of monogalactosylmonoacylglycerols from a freshwater microalgae Chlorella sorokiniana. Nat Prod Res 27: 1028-1031.
50. Kodner RB, Summons RE, Knoll AH (2009) Phylogenetic investigation of the aliphatic, nonhydrolyzable biopolymer algaenan, with a focus on green algae. Org Geochem 40: 854-862.
51. Brand JJ, Wright JN, Lien S (1989) Hydrogen production by eukaryotic algae. Biotechnol Bioeng 33: 1482-1488.
52. Komor E, Cho B-H, Kraus M (1988) The occurrence of the glucose-inducible transport systems for glucose, proline, and arginine in different species of Chlorella. Biotanica Acta 101: 321-326.
53. Lee YK, Ding SY, Hoe CH, Low CS (1996) Mixotrophic growth of Chlorella sorokiniana in outdoor enclosed photobioreactor. J Appl Phycol 8: 163-169. (Pubitemid 26368292)
54. Running JA, Huss RJ, Olson PT (1996) Heterotrophic production of ascorbic acid by microalgae. J Appl Phycol 6: 99-104.
55. Jeong ML, Gillis JM, Hwang J-Y (2003) Carbon dioxide mitigation by microalgal photosynthesis. Bull Korean Chem Soc 24: 1763-1766.
56. Kobayashi N, Noel EA, Barnes A, Watson A, Rosenberg JN, et al. (2013) Characterization of Chlorella sorokiniana strains in anaerobic digested effluent from cattle manure. Bioresource Technol 150: 377-386.
57. Li Y, Zhou W, Hu B, Min M, Chen P, et al. (2011) Integration of algae cultivation as biodiesel production feedstock with municipal wastewater treatment: Strains screening and significance evaluation of environmental factors. Bioresource Technol 102: 10861-10867.
58. Polakovičová G, Kušnír P, Nagyová S, Mikulec J (2012) Process integration of algae production and anaerobic digestion. Chem Eng Trans 29: 1129-1134.
59. Sorokin C, Meyers J (1953) A high-temperature strain of Chlorella . Science 117: 330-331.
60. Fahey R, Buschbacher R, Newton G (1987) The evolution of glutathione metabolism in phototrophic microorganisms. J Mol Evol 25: 81-88.
61. Haehnel W, Nairn JA, Reisberg P, Sauer K (1982) Picosecond fluorescence kinetics and energy transfer in chloroplasts and algae. Biochim Biophys Acta 680: 161-173.
62. Rosen BH, Berliner MD, Petro MJ (1985) Protoplast induction in C. pyrenoidosa. Plant Sci 41: 23-30.
63. 2. Energ Convers Manage 36: 693-696.
64. 13C nuclear magnetic resonance spectroscopy. Appl Environ Microb 54: 1051-1060.
65. Tanaka K, Tomita Y, Tsuruta M, Konishi F, Okuda M, et al. (1990) Oral administration of Chlorella vulgaris augments concomitant antitumor immunity. Immunopharmacol Immunotoxicol 12: 277-291. (Pubitemid 20325688)
66. Wakasugi T, Nagai T, Kapoor M, Sugita M, Ito M, et al. (1997) Complete nucleotide sequence of the chloroplast genome from the green alga Chlorella vulgaris: the existence of genes possibly involved in chloroplast division. Proc Natl Acad Sci USA 94: 5967-5972. (Pubitemid 27241881)
67. Meyer JR, Ellner SP, Hairston NG, Jones LE, Yoshida T (2006) Prey evolution on the time scale of predator-prey dynamics revealed by allele-specific quantitative PCR. Proc Natl Acad Sci USA 103: 10690-10695. (Pubitemid 44078038)
68. Yong-Ha P, Seonggi Y, Mi P, Hoon MS (1997) Livestock waste handling and a high degree of functional biological fertilizer. In: O. . Huimok, editor: Korean Institute of Science and Technology: Biotechnology Institute. Available: http://attfile.konetic.or.kr/konetic/xml/use/31A1D0205268.pdf. Accessed 2014 Mar 1.
69. Guarnieri MT, Nag A, Yang S, Pienkos PT (2013) Proteomic analysis of Chlorella vulgaris: Potential targets for enhanced lipid accumulation. J Proteomics 93: 245-253.
70. Dillon HF, Day AG, Trimbur DE, Im C-S, Franklin S, et al. (2009) Sucrose feedstock utilization for oil-based fuel manufacturing. Patent Number: US8476059. Solazyme, Inc.
71. Dillon HF, Day AG, Trimbur DE, Im C-S, Franklin S, et al. (2009) Glycerol feedstock utilization for oil-based fuel manufacturing. Patent Application Number: US12/194,389. Solazyme, Inc.
72. Wan M, Li P, Xia J, Rosenberg JN, Oyler GA, et al. (2011) The effect of mixotrophy on microalgal growth, lipid content, and expression levels of three pathway genes in Chlorella sorokiniana. Appl Microbiol Biot 91: 835-844.
73. Cooksey KE, Guckert JB, Williams SA, Calli PR (1987) Fluorometric determination of the neutral lipid content of microalgal cells using Nile Red. J Microbiol Meth 6: 333-345.
74. Guckert JB, Cooksey KE (1990) Triglyceride accumulation and fatty acid profile changes in Chlorella (Chlorophyta) during high pH-induced cell cycle inhibition. J Phycol 26: 72-79.
75. Hu Q, Sommerfeld M, Jarvis E, Ghirardi M, Posewitz M, et al. (2008) Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J 54: 621-639. (Pubitemid 351678050)
76. Nguyen HM, Cuiné S, Beyly-Adriano A, Légeret B, Billon E, et al. (2013) The green microalga Chlamydomonas reinhardtii has a single ω-3 fatty acid desaturase which localizes to the chloroplast and impacts both plastidic and extraplastidic membrane lipids. Plant Physiol 113: 223941.
77. Liu J, Sun Z, Zhong Y, Huang J, Hu Q, et al. (2012) Stearoyl-acyl carrier protein desaturase gene from the oleaginous microalga Chlorella zofingiensis: cloning, characterization and transcriptional analysis. Planta 236: 1665-1676.
78. Poerschmann J, Spijkerman E, Langer U (2004) Fatty acid patterns in Chlamydomonas sp. as a marker for nutritional regimes and temperature under extremely acidic conditions. Microb Ecol 48: 78-89. (Pubitemid 39160990)
79. Stephens E, Ross IL, King Z, Mussgnug JH, Kruse O, et al. (2010) An economic and technical evaluation of microalgal biofuels. Nat Biotechnol 28: 126-128.
80. Rogers JN, Rosenberg JN, Guzman BJ, Oh VH, Mimbela LE, et al. (2014) A critical analysis of paddlewheel-driven raceway ponds for algal biofuel production at commercial scales. Algal Res In press.
81. Fan J, Yan C, Zhang X, Xu C (2013) Dual role for phospholipid: diacylglycerol acyltransferase: enhancing fatty acid synthesis and diverting fatty acids from membrane lipids to triacylglycerol in Arabidopsis leaves. Plant Cell Online advance.
82. Prescott GW (1954) How to know the fresh-water algae. Dubuque, IA: C. Brown Company.
83. Bock C, Krienitz L, Pröschold T (2011) Taxonomic reassessment of the genus Chlorella (Trebouxiophyceae) using molecular signatures (barcodes), including description of seven new species. Fottea 11: 293-312.
84. Luo W, Flugmacher SP, Pröschold N, Walz N, Krienitz L (2006) Genotype versus phenotype variability in Chlorella and Micractinium (Chlorophyta, Trebouxiophyceae). Protist 157: 315-333. (Pubitemid 44224242)
85. An SS, Friedl T, Hegewald E (1999) Phylogenetic relationships of Scenedesmus and Scenedesmus-like coccoid green algae as inferred from ITS-2 rDNA sequence comparisons. Plant Biol 1: 418-428. (Pubitemid 29345267)
86. th year. Phycol Res 58: 188-201.
87. Huss VAR, Frank C, Hartmann EC, Hirmer M, Kloboucek A, et al. (1999) Biochemical taxonomy and molecular phylogeny of the genus Chlorella sensu lato (Chlorophyta). J Phycol 35: 587-598. (Pubitemid 29311546)
88. Wan M, Rosenberg JN, Faruq J, Betenbaugh MJ, Xia J (2011) An improved colony PCR procedure for genetic screening of Chlorella and related microalgae. Biotechnol Lett 33: 1615-1619.
89. Noutoshi Y, Ito Y, Kanetani S, Fujie M, Usami S, et al. (1998) Molecular anatomy of a small chromosome in the green alga Chlorella vulgaris . Nucleic Acids Res 26: 3900-3907. (Pubitemid 28401338)
90. Blanc G, Agarkova I, Grinwood J, Kuo A, Brueggeman A, et al. (2012) The genome of the polar eukaryotic microalga Coccomyxa subellipsoidea reveals traits of cold adaptation. Genome Biol 13: R39.
91. Joint Genome Institute (2011) Coccomyxa subellipsoidea C-169 v2.0. U.S. Department of Energy. Available: http://genome.jgi.doe.gov/Coc-C169-1. Accessed 2013 Nov 28.
92. Tirichine L, Bowler C (2011) Decoding algal genomes: tracing back the history of photosynthetic life on Earth. Plant J 66: 45-67.
93. Rosenberg JN, Oyler GA, Wilkinson L, Betenbaugh MJ (2008) A green light for engineered algae: redirecting metabolism to fuel a biotechnology revolution. Curr Opin Biotech 19: 430-436.
94. Campenni L, Nobre BP, Santos CA, Oliveira AC, Aires-Barros MR, et al. (2013) Carotenoid and lipid production by the autotrophic microalga Chlorella protothecoides under nutritional, salinity, and luminosity stress conditions. Appl Microbiol Biot 97: 1383-1393.
95. He PJ, Mao B, Shen CM, Shao LM, Lee DJ, et al. (2013) Cultivation of Chlorella vulgaris on wastewater that contains high levels of ammonia for biodiesel production. Bioresource Technol 129: 177-181.
96. Heredia-Arroyo T, Wei W, Hu B (2010) Oil accumulation via heterotrophic/mixotrophic Chlorella protothecoides. Appl Biochem Biotechnol 162: 1978-1995.
97. Liu ZY, Wang GC, Zhou BC (2008) Effect of iron on growth and lipid accumulation in Chlorella vulgaris. Bioresource Technol 99: 4717-4722.
98. Lu S, Wang J, Niu Y, Yang J, Zhou J, et al. (2012) Metabolic profiling reveals growth related FAME productivity and quality of Chlorella sorokiniana with different inoculum sizes. Biotechnol Bioeng 109: 1651-1662.
99. Santos CA, Ferreira ME, de Silva TL, Gouveia L, Novais JM, et al. (2011) A symbiotic gas exchange between bioreactors enhances microalgal biomass and lipid productivities: taking advantage of complementary nutritional modes. J Ind Microbiol Biotechnol 38: 909-917.
100. Vigeolas H, Duby F, Kaymak E, Niessen G, Motte P, et al. (2012) Isolation and partial characterization of mutants with elevated lipid content in Chlorella sorokiniana and Scenedesmus obliquus. J Biotechnol 162: 3-12.
101. Shen Y, Yuan W, Pei Z, Mao E (2010) Heterotrophic culture of Chlorella protothecoides in various nitrogen sources for lipid production. Appl Biochem Biotechnol 160: 1674-1684.
102. Wu QY, Yin S, Sheng GY, Fu JM (1992) A comparative study of gases generated from stimulant thermal degradation of autotrophic and heterotrophic Chlorella. Prog Nat Sci (in Chinese) 3: 435-440.
103. Kobayashi N, Noel EA, Barnes A, Rosenberg J, DiRusso C, et al. (2013) Rapid detection and quantification of triacylglycerol by HPLC-ELSD in Chlamydomonas reinhardtii and Chlorella strains. Lipids 48: 1035-1049.
104. Wase N, Black PN, Stanley BA, DiRusso CC (2014) Integrated quantitative analysis of nitrogen stress response in Chlamydomonas reinhardtii using metabolite and protein profiling. J Proteome Res, In press.
105. m values and translocation constants of the uptake system. J Gen Physiol 64: 568-581.
106. Ngangkhama M, Rathaa SK, Prasannaa R, Kumarb R, Babua S, et al. (2013) Substrate amendment mediated enhancement of the valorization potential of microalgal lipids. Biocatal Agric Biotechnol 2: 240-246.
107. Haass D, Tanner W (1974) Regulation of hexose transport in Chlorella vulgaris. Plant Physiol 53: 14-20.
108. Kamiya A, Kowallik W (1987) Photoinhibition of glucose uptake in Chlorella. Plant Cell Physiol 28: 617-679.
109. Kim DG, Hur SB (2013) Growth and fatty acid composition of three heterotrophic Chlorella species. Algae 28: 101-109.
110. Reddy HK, Muppaneni T, Rastegary J, Shirazi SA, Ghassemi A, et al. (2013) Hydrothermal extraction and characterization of bio-crude oils from wet Chlorella sorokiniana and Dunaliella tertiolecta . Environ Prog Sust Energy 32: 910-915.
111. Mitra D, van Leeuwen J, Lamsal B (2012) Heterotrophic/mixotrophic cultivation of oleaginous Chlorella vulgaris on industrial co-products. Algal Res 1: 40-48.
112. Nichols HW, Bold HC (1965) Trichosarcina polymorpha Gen. et Sp. Nov. J Phycol 1: 34-38.
113. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, et al. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731-2739.
114. Bligh EG, Dyer WJ (1959) A rapid method for total lipid extraction and purification. Can J Biochem Phys 37: 911-917.
115. Laurens LML, Dempster TA, Jones HDT, Wolfrum EJ, Wychen SV, et al. (2012) Algal biomass constituent analysis: method uncertainties and investigation of the underlying measuring chemistries. Anal Chem 84: 1879-1889.
116. Luthria D, Vinjamoori D, Noel K, Ezzell J (2004) Chapter 3: Accelerated Solvent Extraction. In: D.L . Luthria, editor. Oil Extraction and Analysis: Critical Issues and Comparative Studies. Champaign, IL: American Oil Chemists' Society Press. pp. 25-38.
117. Mulbry W, Kondrad S, Buyer J, Luthria DL (2009) Optimization of an oil extraction process for algae from the treatment of manure effluent. J Am Oil Chem Soc 86: 909-915.