Evaluating renewable carbon sources as substrates for single cell oil production by Cunninghamella echinulata and Mortierella isabellina.

Abstract The biochemical behavior (biomass production, accumulation of total lipid, substrate uptake, fatty acid composition of fungal oil) of two oleaginous Mucorales strains, namely Mortierella isabellina ATHUM 2935 and Cunninghamella echinulata ATHUM 4411, was studied when the aforementioned microorganisms were cultivated on xylose, raw glycerol and glucose under nitrogen-limited conditions. Significant differences in the process of lipid accumulation as related to the carbon sources used were observed for both microorganisms. These differences were attributed to the different metabolic pathways involved in the assimilation of the above substrates. Therefore, the various carbon sources were channeled, at different extent, to storage lipid or to lipid-free biomass formation. Although glucose containing media favored the production of mycelial mass (15 g L −1 of total biomass in the case of C. echinulata and 27 g L −1 in the case of M. isabellina ), the accumulated lipid in dry matter was 46.0% for C. echinulata and 44.6% for M. isabellina . Lipid accumulation was induced on xylose containing media ( M. isabellina accumulated 65.5% and C. echinulata 57.7% of lipid, wt wt −1 , in dry mycelial mass). In these conditions, lipids of C. echinulata contained significant quantities of γ -linolenic acid (GLA). This fungus, when cultivated on xylose, produced 6.7 g L −1 of single cell oil and 1119 mg L −1 of GLA. Finally, the growth of both C. echinulata and M. isabellina on raw glycerol resulted in lower yields in terms of both biomass and oil produced than the growth on xylose.

[1]  J. Folch,et al.  A simple method for the isolation and purification of total lipides from animal tissues. , 1957, The Journal of biological chemistry.

[2]  I. Ellis,et al.  Gamma linolenic acid with tamoxifen as primary therapy in breast cancer , 1998, International journal of cancer.

[3]  A. McAloon,et al.  A process model to estimate biodiesel production costs. , 2006, Bioresource technology.

[4]  R. Fall,et al.  Bioconversion of Xylan to Triglycerides by Oil-Rich Yeasts , 1984, Applied and environmental microbiology.

[5]  E. Belarbi,et al.  Rapid simultaneous lipid extraction and transesterification for fatty acid analyses , 1998 .

[6]  C. T. Evans,et al.  A comparison of the oleaginous yeast,Candida curvata, grown on different carbon sources in continuous and batch culture , 1983, Lipids.

[7]  G. Walker,et al.  Dilute acid hydrolysis of Lignocellulosic biomass , 2010 .

[8]  S. Papanikolaou,et al.  Repression of reserve lipid turnover in Cunninghamella echinulata and Mortierella isabellina cultivated in multiple‐limited media , 2004, Journal of applied microbiology.

[9]  Yoon-Yong Lee,et al.  Dilute-Acid Hydrolysis of Lignocellulosic Biomass , 1999 .

[10]  E. Dumelin Biodiesel - A blessing in disguise? , 2005 .

[11]  Z. Cohen,et al.  Single Cell Oils , 2005 .

[12]  M. Dostálek,et al.  Effect of culture conditions on fatty acid composition in lipids produced by the yeastcryptococcus albidus var. albidus , 1986 .

[13]  S. Papanikolaou,et al.  Accumulation of a Cocoa-Butter-Like Lipid by Yarrowia lipolytica Cultivated on Agro-Industrial Residues , 2003, Current Microbiology.

[14]  Z. Zong-bao,et al.  Screening of Oleaginous Yeasts for Broad-Spectrum Carbohydrates Assimilating Capacity , 2005 .

[15]  A. H. Romano Microbial sugar transport systems and their importance in biotechnology , 1986 .

[16]  W. Rymowicz,et al.  Citric acid production from raw glycerol by acetate mutants of Yarrowia lipolytica , 2006 .

[17]  F. Shewmaker,et al.  γ-Linolenic acid in zygomycetous fungi: Syzygites megalocarpus , 1998 .

[18]  S. Narine,et al.  Implications for the use of Mortierella fungi in the industrial production of essential fatty acids , 2005 .

[19]  S. Narine,et al.  Maximizing the production of γ-linolenic acid in Mortierella ramanniana var. ramanniana as a function of pH, temperature and carbon source, nitrogen source, metal ions and oil supplementation , 2005 .

[20]  M. Dostálek,et al.  Effect of culture conditions on mycelial growth and production of γ-linolenic acid by the fungus Mortierella ramanniana , 1988, Applied Microbiology and Biotechnology.

[21]  Seraphim Papanikolaou,et al.  Single cell oil (SCO) production by Mortierella isabellina grown on high-sugar content media. , 2004, Bioresource technology.

[22]  G. L. Miller Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar , 1959 .

[23]  Hung-Chang Chen,et al.  Production of γ‐Linolenic Acid by the Fungus Cunninghamella echinulata CCRC 31840 , 1996 .

[24]  Wei Wang,et al.  Microbial production of 1,3-propanediol by Klebsiella pneumoniae using crude glycerol from biodiesel preparations , 2006, Biotechnology Letters.

[25]  C. Ratledge Single Cell Oils for the 21st Century , 2010 .

[26]  昌 清水,et al.  Single cell oil , 1982 .

[27]  Fengwu Bai,et al.  High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture , 2007 .

[28]  C Ratledge,et al.  Biochemical events leading to the diversion of carbon into storage lipids in the oleaginous fungi Mucor circinelloides and Mortierella alpina. , 2001, Microbiology.

[29]  M. Dostálek,et al.  Production of γ-linolenic acid by the fungus Mucor rouxii in fed-batch and continuous culture , 1989, Applied Microbiology and Biotechnology.

[30]  Seraphim Papanikolaou,et al.  Lipid production by Yarrowia lipolytica growing on industrial glycerol in a single-stage continuous culture. , 2002, Bioresource technology.

[31]  Hui Luo,et al.  A new method for preparing raw material for biodiesel production , 2006 .

[32]  S. Papanikolaou,et al.  Biotechnological valorisation of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: Production of 1,3-propanediol, citric acid and single cell oil , 2008 .

[33]  M. Komaitis,et al.  Production of γ-linolenic acid by Cunninghamella echinulata cultivated on glucose and orange peel , 2002, Applied Microbiology and Biotechnology.

[34]  G. Eggink,et al.  High-cell-density cultivation of the lipid accumulating yeast Cryptococcus curvatus using glycerol as a carbon source , 1996, Applied Microbiology and Biotechnology.

[35]  Seraphim Papanikolaou,et al.  Modelling aspects of the biotechnological valorization of raw glycerol: production of citric acid by Yarrowia lipolytica and 1,3‐propanediol by Clostridium butyricum , 2003 .

[36]  Sudheer Kumar Singh,et al.  Effects of various process parameters on the production of gamma-linolenic acid in submerged fermentation , 2006 .

[37]  C. T. Evans,et al.  Induction of xylulose-5-phosphate phosphoketolase in a variety of yeasts grown ond-xylose: the key to efficient xylose metabolism , 1984, Archives of Microbiology.

[38]  S. Papanikolaou,et al.  Lipids of Cunninghamella echinulata with emphasis to γ-linolenic acid distribution among lipid classes , 2006, Applied Microbiology and Biotechnology.