Engineering of lipid production in oleaginous microorganisms

Subject Areas : Microbial Biotechnology

Fateme Ghanaatian ¹ , Ahmad Farhad Talebi ²

1 - M.Sc. student, Faculty of Biotechnology, Semnan University, Semnan, Iran.
2 - Assistant Professor, Faculty of Biotechnology, Semnan University, Semnan, Iran.

Received: 2017-09-20 Accepted : 2017-10-02 Published : 2018-05-22

Keywords: Biodiesel, lipid metabolism, Genetic Engineering, Oily microorganisms, Microbial oils,

Abstract :

Microbial oils are of great interest to researchers as they provide essential fatty acids and are among renewable energy sources. Oleaginous microorganisms have the ability to produce oils up to 60% of their weight, most of which are accumulated in the form of triglycerides. Fouroleaginous genera including bacteria, microalgae, fungi, and yeasts are among the largestproducers of microbial oils. A variety of physical and chemical factors are effective in microbial oils production, among which carbon source, nutrient deficiency, temperature, light intensity and pH of the environment can be noticed. Considering the constraints faced by various physical and chemical treatments in microbial oils production, most researches are currently focusing ongenetic modifications to enhance lipid production in oleaginous microorganisms. This articlereviewed 210 peer-reviewed articles entitled engineering of lipid production in oleaginousmicroorganisms in scientific databases during 1990-2017, among which 89 to particles wereselected for further assessments. In order to have the best achievement in replacement of new oil sources on a large scale, these resources should be optimized in terms of unique properties such as unsaturated fatty acids production, lipid yield, as well as lipid profile. The present study is anattempt to recommend some basic approaches to increase microbial oils production by oleaginous microorganisms. These strategies should be in line with genetic and environment-protection priorities. Concurrently, they could be useful for knowledge-based enterprises from the economical point of view.

References:

1. Yen HW, Hu IC, Chen CY, Ho SH, Lee DJ, Chang JS. Microalgae-based biorefinery from
   biofuels to natural products. Bioresour Technol. 2013; 135: 166-174.
   2. Talebi AF, Mohtashami SK, Tabatabaei M, Tohidfar M, Bagheri A, Zeinalabedini M, Mirzaei
   H, Mirzajanzadeh M, Shafaroudi S, Bakhtiari S. Fatty acids profiling: a selective criterion for
   screening microalgae strains for biodiesel production. Algal Res. 2013; 2(3): 258-267.
   3. Liang MH, Jiang JG. Advancing oleaginous microorganisms to produce lipid via metabolic
   engineering technology. Prog Lipid Res. 2013; 52(4): 395-408.
   4. Bellou S, Triantaphyllidou IE, Aggeli D, Elazzazy AM, Baeshen MN, Aggelis G. Microbial
   oils as food additives: recent approaches for improving microbial oil production and its
   polyunsaturated fatty acid content. Curr Opin Biotechnol. 2016; 37: 24-35.
   5. Talebi AF, Tohidfar M, Mousavi Derazmahalleh SM, Sulaiman A, Baharuddin AS, Tabatabaei
   M. Biochemical modulation of lipid pathway in microalgae Dunaliella sp. for biodiesel
   production. Biomed Res Int. 2015; 2015: 1-12.
   6. Donot F, Fontana A, Baccou J, Strub C, Schorr-Galindo S. Single cell oils (SCOs) from
   oleaginous yeasts and moulds: production and genetics. Biomass Bioenergy. 2014; 68:
   135-150.
   7. Adrio JL. Oleaginous yeasts: promising platforms for the production of oleochemicals and
   biofuels. Biotechnol Bioeng. 2017; 144(19): 1915-1920.
   8. Beopoulos A, Nicaud J-M, Gaillardin C. An overview of lipid metabolism in yeasts and its
   impact on biotechnological processes. Appl Microbiol Biotechnol. 2011; 90(4): 1193-1206.
   9. Bazinet RP, Layé S. Polyunsaturated fatty acids and their metabolites in brain function and
   disease. Nat Rev Neurosci. 2014; 15(12): 771-778.
   10. Bruen R, Fitzsimons S, Belton O. Atheroprotective effects of conjugated linoleic acid. Br J
   Clin Pharmacol. 2017; 83(1): 46-53.
   11. Passoth V. Lipids of yeasts and filamentous fungi and their importance for biotechnology.
   Biotechnology of Yeasts and Filamentous Fungi: Springer; 2017; 83(1): 149-204.
   12. ača , arcinčá , Čertí , pel a , arcinčá á , t á , lnár ,
   lemp á , as aľ á . ffect f a in prefrmente cereal pr ct c ntainin
   gamma-linolenic acid to broiler feed on production indicators and fatty acid profile of chicken
   breast. Acta Ve Brno. 2014; 83(4): 379-384.
   13. Madani M, Enshaeieh M, Abdoli A. Single cell oil and its application for biodiesel
   production. Process Saf Environ. 2017; 92(3): 46-53.
   14. Li Q, Du W, Liu D. Perspectives of microbial oils for biodiesel production. Appl Microbiol
   Biotechnol. 2008; 80(5): 749-756.
   15. Spolaore P, Joannis-Cassan C, Duran E, Isambert A. Commercial applications of microalgae.
   J Biosci Bioeng. 2006; 101(2): 87-96.
   
   16. Odjadjare EC, Mutanda T, Olaniran AO. Potential biotechnological application of microalgae:
   a critical review. Crit Rev Biotechnol. 2017; 37(1): 37-52.
   17. Shaojin Y, Yiping Z. Research and application of oleaginous microorganism. China
   Foreign Energy. 2006; 2: 21-32.
   18. Xue F, Zhang X, Tan T. Research advance and prospect in microbial oils. Chinese J
   Bioprocess Engin. 2005; 3(1): 23-27.
   19. Gouda MK, Omar SH, Aouad LM. Single cell oil production by Gordonia sp. DG using
   agro-industrial wastes. World J Microbiol Biotechnol. 2008; 24(9): 1703-1659.
   20. Ratledge C. Fatty acid biosynthesis in microorganisms being used for single cell oil
   production. Biochem. 2004; 86(11): 807-815.
   21. Haslam TM, Kunst L. Extending the story of very-long-chain fatty acid elongation. Plant Sci.
   2013; 210: 93-107.
   22. Koolman J, Röhm KH, Wirth J, Robertson M. Color atlas of biochemistry. Thieme Stuttgart;
   2005; 120: 120-152.
   23. Berg JM, Tymoczko JL. Biochemistry/Jeremy M. Berg, John L. Tymoczko, Lubert Stryer;
   web content by Neil D. Clarke. 1998; pp: 524-560.
   24. Zhang JY, Kothapalli KS, Brenna JT. Desaturase and elongase-limiting endogenous
   long-chain polyunsaturated fatty acid biosynthesis. Curr Opin Clin Nutr. 2016; 19(2): 103-110.
   25. Kosa M, Ragauskas AJ. Lipids from heterotrophic microbes: advances in metabolism
   research. Trends Biotechnol. 2011; 29(2): 53-61.
   26. Rajakumari S, Grillitsch K, Daum G. Synthesis and turnover of non-polar lipids in yeast. Prog
   Lipid Res. 2008; 47(3): 157-171.
   27. Courchesne NMD, Parisien A, Wang B, Lan CQ. Enhancement of lipid production using
   biochemical, genetic and transcription factor engineering approaches. J Biotechnol. 2009; 141
   (1): 31-41.
   28. Talebi AF, Tohidfar M, Bagheri A, Lyon SR, Salehi-Ashtiani k, Tabatabaei M. Manipulation
   of carbon flux into fatty acid biosynthesis pathway in Dunaliella salina using AccD and ME
   genes to enhance lipid content and to improve produced biodiesel quality. Biofuel Res J. 2014;
   1(3): 91-97.
   29. Yang S, Wang W, Wei H, Van Wychen S, Pienkos PT, Zhang M, Himmel M. Comparison of
   nitrogen depletion and repletion on lipid production in yeast and fungal species. Energies.
   2016; 9(9): 685-699.
   30. Subramaniam R, Dufreche S, Zappi M, Bajpai R. Microbial lipids from renewable resources:
   production and characterization. J Ind Microbiol Biotechnol. 2010; 37(12): 1271-1287.
   31. Ruenwai R, Cheevadhanarak S, Laoteng K. Overexpression of acetyl-CoA carboxylase gene
   of Mucor rouxii enhanced fatty acid content in Hansenula polymorpha. Mol Biotechnol. 2009;
   42(3): 327-332.
   32. Zhang S, Skerker JM, Rutter CD, Maurer MJ, Arkin AP, Rao CV. Engineering
   
   Rhodosporidium toruloides for increased lipid production. Biotechnol Bioeng. 2016; 113(5):
   1056-1066.
   33. Feng D, Chen Z, Xue S, Zhang W. Increased lipid production of the marine oleaginous
   microalgae Isochrysis zhangjiangensis (Chrysophyta) by nitrogen supplement. Bioresour.
   Technol. 2011; 102(12): 6710-6716.
   34. Khozin-Goldberg I, Cohen Z. The effect of phosphate starvation on the lipid and fatty acid
   composition of the fresh water eustigmatophyte Monodus subterraneus. Phytochemistry. 2006;
   67(7): 696-701.
   35. Lei A, Chen H, Shen G, Hu Z, Chen L, Wang J. Expression of fatty acid synthesis genes and
   fatty acid accumulation in Haematococcus pluvialis under different stressors. Biotechnol
   Biofuels. 2012; 5(1): 1-25.
   36. Li Y, Han D, Hu G, Sommerfeld M, Hu Q. Inhibition of starch synthesis results in
   overproduction of lipids in Chlamydomonas reinhardtii. Biotechnol Bioeng. 2010; 107(2):
   258-268.
   37. Arabolaza A, Rodriguez E, Altabe S, Alvarez H, Gramajo H. Multiple pathways for
   triacylglycerol biosynthesis in Streptomyces coelicolor. Appl Environ Microbiol. 2008; 74(9):
   2573-2582.
   38. Kalscheuer R, Stöveken T, Malkus U, Reichelt R, Golyshin PN, Sabirova JS, Ferrer M,
   Timmis K, Steinbüchel A. Analysis of storage lipid accumulation in Alcanivorax borkumensis:
   evidence for alternative triacylglycerol biosynthesis routes in bacteria. J Bacteriol. 2007; 189
   (3): 918-928.
   39. Zhang X, Li M, Agrawal A, San KY. Efficient free fatty acid production in Escherichia coli
   using plant acyl-ACP thioesterases. Metab Eng. 2011; 13(6): 713-722.
   40. Fakas S, Papanikolaou S, Batsos A, Galiotou-Panayotou M, Mallouchos A, Aggelis G.
   Evaluating renewable carbon sources as substrates for single cell oil production by
   Cunninghamella echinulata and Mortierella isabellina. Biomass Bioenergy. 2009; 33(4):
   573-580.
   41. Čertí , altesz , Ša bi r J. ipi f rmati n an γ-linolenic acid production by
   Mucorales fungi grown on sunflower oil. Lett Appl Microbiol. 1997; 25(2): 101-105.
   42. Zhu L, Zong M, Wu H. Efficient lipid production with Trichosporonfermentans and its use for
   biodiesel preparation. Bioresour Technol. 2008; 99(16): 7881-7885.
   43. Xu J, Du W, Zhao X, Zhang G, Liu D. Microbial oil production from various carbon sources
   and its use for biodiesel preparation. Biofuel Bioprod Bior. 2013; 7(1): 65-77.
   44. Cheirsilp B, Kitcha S, Torpee S. Co-culture of an oleaginous yeast Rhodotorula glutinis and a
   microalga Chlorella vulgaris for biomass and lipid production using pure and crude glycerol as
   a sole carbon source. Ann microbiol. 2012; 62(3): 987-993.
   45. Kraisintu P, Yongmanitchai W, Limtong S. Selection and optimization for lipid production of
   a newly isolated oleaginous yeast, Rdodosporidium toruloides DMKU3-TK16. Kasetsart J Nat
   
   Sci. 2010; 44: 436-445.
   46. Xin L, Hong-ying H, Ke G, Ying-xue S. Effects of different nitrogen and phosphorus
   concentrations on the growth, nutrient uptake, and lipid accumulation of a freshwater
   microalga Scenedesmus sp. Bioresour Technol. 2010; 101(14): 5494-5500.
   47. Tang S, Chen M, Yang J, Ni Q, He D, Chen T. Research of producing oil by Mortierella
   isabellina. China Oils Fats. 2007; 32(12): 35-37.
   48. Wu S, Hu C, Jin G, Zhao X, Zhao ZK. Phosphate-limitation mediated lipid production by
   Rhodosporidium toruloides. Bioresour Technol. 2010; 101(15): 6124-6129.
   49. Esakkimuthu S, Krishnamurthy V, Govindarajan R ,Swaminathan K. Augmentation and
   starvation of calcium, magnesium, phosphate on lipid production of Scenedesmus obliquus.
   Biomass Bioenergy. 2016; 88: 126-134.
   50. Iwai M, Hori K, Sasaki-Sekimoto Y, Shimojima M, Ohta H. Manipulation of oil synthesis in
   Nannochloropsis strain NIES-2145 with a phosphorus starvation–inducible promoter from
   Chlamydomonas reinhardtii. Front Microbiol. 2015; 6: 912-918.
   51. Mizuno Y, Sato A, Watanabe K, Hirata A, Takeshita T, Ota S, Sato N, Zachleder V, Tsuzuki
   M, Kawano S. Sequential accumulation of starch and lipid induced by sulfur deficiency in
   Chlorella and Parachlorella species. Bioresour Technol. 2013; 129: 150-155.
   52. Sun X, Cao Y, Xu H, Liu Y, Sun J, Qiao D, Cao Y. Effect of nitrogen-starvation, light
   intensity and iron on triacylglyceride/carbohydrate production and fatty acid profile of
   Neochloris oleoabundans HK-129 by a two-stage process. Bioresour Technol. 2014; 155:
   204-212.
   53. 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(3): 3034-3040.
   54. Kim JH, Choi SK, Park YS, Yun C-W, Cho WD, Chee KM, Chang H. Effect of culture
   conditions on astaxanthin formation in red yeast Xanthophyllomyces dendrorhous mutant JH1.
   J Microbiol Biotechnol. 2006; 16(3): 438-442.
   55. Kruszewska J, Palamarczyk G, Kubicek CP. Stimulation of exoprotein secretion by choline
   and Tween 80 in Trichoderma reesei QM 9414 correlates with increased activities of dolichol
   phosphate mannose synthase. Microbiol. 1990; 136(7): 1293-1298.
   56. Dalmau E, Montesinos J, Lotti M, Casas C. Effect of different carbon sources on lipase
   production by Candida rugosa. Enzyme Microb Technol. 2000; 26(9): 657-663.
   57. Saenge C, Cheirsilp B, Suksaroge TT, Bourtoom T. Efficient concomitant production of lipids
   and carotenoids by oleaginous red yeast Rhodotorula glutinis cultured in palm oil mill effluent
   and application of lipids for biodiesel production. Biotechnol Bioprocess Eng. 2011; 16(1):
   23-33.
   58. Ageitos JM, Vallejo JA, Veiga-Crespo P, Villa TG. Oily yeasts as oleaginous cell factories.
   Appl Microbiol Biotechnol. 2011; 90(4): 1219-1227.
   
   59. Beltran G, Novo M, Guillamón JM, Mas A, Rozès N. Effect of fermentation temperature and
   culture media on the yeast lipid composition and wine volatile compounds. Int J Food
   Microbiol. 2008; 121(2): 169-177.
   60. Sharma KK, Schuhmann H, Schenk PM. High lipid induction in microalgae for biodiesel
   production. Energies. 2012; 5(5): 1532-1553.
   61. Angerbauer C, Siebenhofer M, Mittelbach M, Guebitz G. Conversion of sewage sludge into
   lipids by Lipomyces starkeyi for biodiesel production. Bioresour Technol. 2008; 99(8):
   3051-3056.
   62. Einicker-Lamas M, Mezian GA, Fernandes TB, Silva FLS, Guerra F, Miranda K, Attias M,
   Oliveira M. Euglena gracilis as a model for the study of Cu2+ and Zn2+ toxicity and
   accumulation in eukaryotic cells. Environ Pollut. 2002; 120(3): 779-786.
   63. Liu ZY, Wang GC, Zhou BC. Effect of iron on growth and lipid accumulation in Chlorella
   vulgaris. Bioresour Technol. 2008; 99(11): 17-22.
   64. Yen HW, Zhang Z. Enhancement of cell growth rate by light irradiation in the cultivation of
   Rhodotorula glutinis. Bioresour Technol. 2011; 102(19): 9279-9281.
   65. Nielsen J. Synthetic biology for engineering acetyl coenzyme A metabolism in yeast. MBio.
   2014; 5(6): 2141-2153.
   66. Tai M, Stephanopoulos G. Engineering the push and pull of lipid biosynthesis in oleaginous
   yeast Yarrowia lipolytica for biofuel production. Metab Eng. 2013; 15: 1-9.
   67. Runguphan W, Keasling JD. Metabolic engineering. of Saccharomyces cerevisiae for
   production of fatty acid-derived biofuels and chemicals. Metab Eng. 2014; 21: 3-13.
   68. Lu X, Vora H, Khosla C. Overproduction of free fatty acids in E. coli: implications for
   biodiesel production. Metab Eng. 2008; 10(6): 333-339.
   69. Voelker TA, Davies HM. Alteration of the specificity and regulation of fatty acid synthesis of
   Escherichia coli by expression of a plant medium-chain acyl-acyl carrier protein thioesterase. J
   Bacteriol. 1994; 176(23): 7320-7327.
   70. Zheng Z, Zou J. The initial step of the glycerolipid pathway identification of glycerol
   3-phosphate/dihydroxyacetone phosphate dual substrate acyltransferases in Saccharomyces
   cerevisiae. J Biol Chem. 2001; 276(45): 14710-14716.
   71. Zaremberg V, McMaster CR. Differential partitioning of lipids metabolized by separate yeast
   glycerol-3-phosphate acyltransferases reveals that phospholipase D generation of phosphatidic
   acid mediates sensitivity to choline-containing lysolipids and drugs. J Biol Chem. 2002; 277
   (41): 39035-39044.
   72. Olukoshi ER, Packter NM. Importance of stored triacylglycerols in Streptomyces: possible
   carbon source for antibiotics. Microbiol. 1994; 140(4): 931-943.
   73. Qiao K, Abidi SHI, Liu H, Zhang H, Chakraborty S, Watson N, Ajikumar P, Stephanopoulos
   G. Engineering lipid overproduction in the oleaginous yeast Yarrowia lipolytica. Metab Eng.
   2015; 29: 56-65.
   
   74. Vigeolas H, Waldeck P, Zank T, Geigenberger P. Increasing seed oil content in oil-seed rape
   (Brassica napus L.) by over‐expression of a yeast glycerol‐3‐phosphate dehydrogenase
   under the control of a seed‐specific promoter. Plant Biotechnol J. 2007; 5(3): 431-441.
   75. Beopoulos A, Mrozova Z, Thevenieau F, Le Dall M-T, Hapala I, Papanikolaou S, Chardot T,
   Nicaud J. Control of lipid accumulation in the yeast Yarrowia lipolytica. Appl Environ
   Microbiol. 2008; 74; 7779-7789.
   76. Dulermo T, Nicaud J-M. Involvement of the G3P shuttle and β-oxidation pathway in the
   control of TAG synthesis and lipid accumulation in Yarrowia lipolytica. Metab Eng. 2011; 13
   (5): 482-491.
   77. Pietrocola F, Galluzzi L, Bravo-San Pedro JM, Madeo F, Kroemer G. Acetyl coenzyme A: a
   central metabolite and second messenger. Cell Metab. 2015; 21(6): 805-821.
   78. Lin H, Castro NM, Bennett GN, San K-Y. Acetyl-CoA synthetase overexpression in
   Escherichia coli demonstrates more efficient acetate assimilation and lower acetate
   accumulation: a potential tool in Metabolic engineering. Appl Microbiol Biotechnol. 2006; 71
   (6): 870-874.
   79. Ratledge C. The role of malic enzyme as the provider of NADPH in oleaginous
   microorganisms: a reappraisal and unsolved problems. Biotechnol Lett. 2014; 36(8):
   1557-1568.
   80. Hou L, Shi D, Cai Z, Song D, Wang X. Regulation of lipids synthesis in transgenic
   Escherichia coli by inserting Cyanobacterial sense and antisense pepcA gene. China Biotech
   2008; 52: 25-28.
   81. Meng X, Yang J, Cao Y, Li L, Jiang X, Xu X, Liu W, Xian M, Zhang Y. Increasing fatty acid
   production in E. coli by simulating the lipid accumulation of oleaginous microorganisms. J Ind
   Microbiol Biotechnol. 2011; 38(8): 919-925.
   82. Beopoulos A, Cescut J, Haddouche R, Uribelarrea JL, Molina-Jouve C, Nicaud JM.
   Yarrowia lipolytica as a model for bio-oil production. Prog Lipid Res. 2009; 48(6): 375-387.
   83. Tamano K, Bruno KS, Karagiosis SA, Culley DE, Deng S, Collett JR, Umemura M, Koike H,
   Baker S, Machida M. Increased production of fatty acids and triglycerides in Aspergillus
   oryzae by enhancing expressions of fatty acid synthesis-related genes. Appl Microbiol
   Biotechnol. 2013; 97(1): 269-281.
   84. Scharnewski M, Pongdontri P, Mora G, Hoppert M, Fulda M. Mutants of Saccharomyces
   cerevisiae deficient in acyl‐CoA synthetases secrete fatty acids due to interrupted fatty acid
   recycling. FEBS J. 2008; 275(11): 2765-2778.
   85. Beopoulos A, Chardot T, Nicaud J-M. Yarrowia lipolytica: A model and a tool to understand
   the mechanisms implicated in lipid accumulation. Biochem. 2009; 91(6): 692-696.
   86. líč á , , t enstae t , ' n rea , a m , ar t , Nica J. ipi
   accumulation, lipid body formation, and acyl coenzyme A oxidases of the yeast Yarrowia
   lipolytica. Appl Environ Microbiol. 2004; 70(7): 3918-3924.
   
   87. Coleman RA, Lee DP. Enzymes of triacylglycerol synthesis and their regulation. Prog Lipid
   Res. 2004; 43(2): 134-176.
   88. Todd BL, Stewart EV, Burg JS, Hughes AL, Espenshade PJ. Sterol regulatory element
   binding protein is a principal regulator of anaerobic gene expression in fission yeast. Mol Cell
   Biol. 2006; 26(7): 2817-2831.
   89. Zhang B, Chen H, Li M, Gu Z, Song Y, Ratledge C, Chen Y, Zhang H, Chen W. Genetic
   engineering of Yarrowia lipolytica for enhanced production of trans-10, cis-12 conjugated
   linoleic acid. Microb Cell Fact. 2013; 12: 70.
   90. Najafi G, Ghobadian B, Tavakoli T, Yusaf T. Potential of bioethanol production from
   agricultural wastes in Iran. Renew Sust Energ Rev. 2009; 13(6): 1418-1427.
   91. Tabatabaei M, Tohidfar M, Jouzani GS, Safarnejad M, Pazouki M. Biodiesel production from
   genetically engineered microalgae: future of bioenergy in Iran. Renew Sust Energ Rev. 2011;
   15(4): 1918-1927.
   92. Raeesossadati MJ, Ahmadzadeh H, McHenry MP, Moheimani NR. CO2 bioremediation by
   microalgae in photobioreactors: impacts of biomass and CO2 concentrations, light, and
   temperature. Algal Res. 2014; 6: 78-85.
   93. Madadi R, Pourbabaee AA, Tabatabaei M, Zahed MA, Naghavi MR. Treatment of
   petrochemical wastewater by the green algae Chlorella vulgaris. Int J Environ Res. 2016; 10
   (4): 555-560.
   94. Najafi G, Ghobadian B, Yusaf TF. Algae as a sustainable energy source for biofuel production
   in Iran: a case study. Renew Sustainable Energy Rev. 2011; 15(8): 3870-3876.
   95. Abdoli A, Enshaeieh M, Nahvi I, Madani M. Isolation of oleaginous yeasts and optimization
   of lipid production using taguchi design. New Cell Mol Biotechnol J. 2012; 4(14): 13-20.
   96. Ghasemi L, Samadlouie H, Jalali H, Gharanjik S. Isolation and identification of Candida
   orthopsilosi SAGSGC as oleaginous yeast in perch fish by using ribosomal gene and
   optimization of oil and biomass production. J Agr Food Chem. 2017; 14(70): 1-12.
   97. Pourbabaee A, Mondaniizadeh M. Single cell oil production from petroleum sludge by
   native yeast strains. J Rene Energy Environ. 2014; 2: 19.
   98. Enshaeieh M, Abdoli A, Madani M. Single cell oil (SCO) production by Rhodotorula
   mucilaginosa and its environmental benefits. J Agr Sci Tech. 2015; 17(2): 387-400.
   99. Enshaeieh M, Abdoli A, Nahvi I, Madani M. Bioconversion of different carbon sources in to
   microbial oil and biodiesel using oleaginous yeasts. J Biol Todays World. 2012; 1(2): 82-92.
   100. Shafiei N, Beheshti MK, Madani M. Isolation, optimization, and investigation of production
   of linoleic acid in Aspergillus niger. Qom Univ Med Sci J. 2016; 10(6): 24-31.
   101. Mohammadi Nasr M, Nahvi I, Biria D, Mirbagheri M. Optimization of culture media for
   enhancing gamma-linolenic acid production by Mucor hiemalis. Biological J Microorganism.
   2016; 4(16): 25-31.
   102. Nasr MM, Nahvi I, Keyhanfar M, Mirbagheri M. The effect of carbon and nitrogen sources
   on the fatty acids profile of Mortierella vinacea. Biological J Microorganism. 2017; 5(20): 1