Comparative lipidomic analysis of S. cerevisiae cells during industrial bioethanol fermentation

Abstract

Variations in the composition and level of phospholipids (PLs) in yeast cells during industrial ethanol fermentation processes were analyzed. A comparative lipidomic method was used to investigate the changes in total cellular PLs during continuous and fed-batch/batch processes. The phospholipid metabolism in yeast changed during both processes, mainly due to the presence of long-chain poly unsaturated fatty acids (PUFA) that contained phosphatidyglycerol (PG), phosphatidylethanolamine (PE) and phosphatidylserine (PS). The complexity of the media affected the growth of the yeast and the membrane composition. Yeast incorporated lots of exogenous saturated and PUFAs from the feedstock during the fermentations. During the continuous fermentation, there was an increase in PLs with shorter chains as the fermentation progressed and early in process there were more long-chains. During the fed-batch/batch process, the PG species increased as the fermentation progressed. This is probably due to an inositol deficiency in the earlier part of the fermentation.

Keywords

lipidomics / systems biology / phospholipids / stirred-tank / saccharomyces cerevisiae / biorefinery engineering

References

[1] Ikner A, Shiozaki K.Yeast signaling pathways in the oxidative stress response mutation research/fundamental and molecular mechanisms of mutagenesis, 2005, 569(1–2): 13–27

[2] Yamaji K, Hara S, Mizoguchi H. Influence of Ras function on ethanol stress response of sake yeast. Journal of Bioscience and Bioengineering, 2003, 96(5): 474–480

[3] Gasch A P, Spellman P T, Kao C M, Carmel-harel O, Eisen M B, Storz G, Botstein D, Brown P O. Genomic expression programs in the response of yeast cells to environmental changes. Molecular Biology of the Cell, 2000, 11: 4241–4257

[4] Puig S, Perez-Ortin J E. Expression levels and patterns of glycolytic yeast genes during wine fermentation. Original Research Article Systematic and Applied Microbiology, 2000, 23(2): 300–303

[5] Beltran G, Novo M, Guillamon J M, Mas A, Rozes N. Effect of fermentation temperature and culture media on the yeast lipid composition and wine volatile compounds. International Journal of Food Microbiology, 2008, 121(2): 169–177

[6] Torija M J, Beltran G, Novo M, Poblet M, Guillamon J M, Mas A, Rozes N. Effects of fermentation temperature and Saccharomyces species on the cell fatty acid composition and presence of volatile compounds in wine. International Journal of Food Microbiology, 2003, 85(1–2): 127–136

[7] Xu T J, Zhao X Q, Bai F W. Continuous ethanol production using self-flocculating yeast in a cascade of fermentors. Enzyme and Microbial Technology, 2005, 37(6): 634–640

[8] Tang Y Q, An M Z, Zhong Y L, Shigeru M, Wu X L, Kida K. Continuous ethanol fermentation from non-sulfuric acid-washed molasses using traditional stirred tank reactors and the flocculating yeast strain KF-7. Original Journal of Bioscience and Bioengineering, 2010, 109(1): 41–46

[9] Alfenore S, Cameleyre X, Benbadis L, Bideaux C, Uribelarrea J L, Goma G, Molina-Jouve C, Guillouet S E. Aeration strategy: a need for very high ethanol performance in saccharomyces cerevisiae fed-batch process. Applied Microbiology and Biotechnology, 2004, 63(5): 537–542

[10] Laluce C, Souza C S, Abud C L, Gattas E A L, Walker G M. Continuous ethanol production in a nonconventional five-stage system operating with yeast cell recycling at elevated temperatures. Journal of Industrial Microbiology & Biotechnology, 2002, 29(3): 140–144

[11] Hojo O, Hokka C O, Ana M M S. Ethanol production by a flocculant yeast strain in a CSTR type fermentor with cell recycling. Applied Microbiology and Biotechnology, 1999, 78(1–3): 535–545

[12] Mannazzu I, Angelozzi D, Belviso S, Budroni M, Farris G A, Goffrini P, Lodi T, Marzona M, Bardi L. Behaviour of saccharomyces cerevisiae wine strains during adaptation to unfavourable conditions of fermentation on synthetic medium: cell lipid composition, membrane integrity, viability and fermentative activity. International Journal of Food Microbiology, 2008, 121(1): 84–91

[13] Benchekroun K, Bonaly R. Physiological properties and plasma membrane composition of saccharomyces cerevisiae grown in sequential batch culture and in presence of surfactant. Applied Microbiology and Biotechnology, 1992, 36(5): 673–678

[14] Boumann H A, Damen M J A, Versluis C, Heck A J R, de Kruijff B, de Kroon A. The two biosynthetic routes leading to phosphatidylcholine in yeast produce different sets of molecular species. Evidence for lipid remodeling. Biochem, 2003, 42(10): 3054–3059

[15] Garnier M, Dufourc E J, Larijani B. Characterisation of lipids in cell signaling and membrane dynamics by nuclear magnetic resonance spectroscopy and mass spectrometry. Signal Transduction, 2006, 6(2): 133–143

[16] Nikolic M, Stanic D, Baricevic I, Jones D R, Nedic O, Niketic V. Efflux of cholesterol and phospholipids derived from the haemoglobin-lipid adduct in human red blood cells into plasma. Clinical Biochemistry, 2007, 40(5–6): 305–309

[17] Czabany T, Athenstaedt K, Daum G. Synthesis, storage and degradation of neutral lipids in yeast. Biochimica et Biophysica Acta (BBA)-. Molecular and Cell Biology of Lipids, 2007, 1771(3): 299–309

[18] Takeshi K, Kazuo S. Disruption of a gene encoding phosphatidic acid phosphatase causes abnormal phenotypes in cell growth and abnormal cytokinesis in saccharomyces cerevisiae. Biochemical and Biophysical Research Communications, 1998, 248(1): 87–92

[19] Vance J E, Steenbergen R. Metabolism and functions of phosphatidylserine. Progress in Lipid Research, 2005, 44(4): 207–234

[20] Choi J Y, Martin W E, Murphy R C, Voelker D R. Phosphatidylcholine and N-methylated phospholipids are nonessential in saccharomyces cerevisiae. Journal of Biological Chemistry, 2004, 279(40): 42321–42330

[21] Chen S, Li K W. Comparison of molecular species of various transphosphatidylated phosphatidylserine (PS) with bovine cortex PS by mass spectrometry. Chemistry and Physics of Lipids, 2008, 152(1): 46–56

[22] Yang S, Lu S H, Yuan Y J. Lipidomics analysis reveals different defense responses of Taxus cuspidate cells to two elicitors, methyljasmonate and cerium (Ce⁴⁺). Biochimica et Biophysica Acta (BBA)-. Molecular and Cell Biology of Lipids, 2008, 1781(3): 123–134

[23] Karin A Z B, Robert C M. Electrospray ionization tandem mass spectrometry of glycerophosphoethanolamine plasmalogen phospholipids. Journal of the American Society for Mass Spectrometry, 2004, 15(10): 1499–1508

[24] Wang C, Xie S, Yang J, Yang Q, Xua G. Structural identification of human blood phospholipids using liquid chromatography/quadrupole-linear ion trap mass spectrometry. Analytica Chimica Acta, 2004, 525(1): 1–10

[25] Cheng J S, Qiao B, Yuan Y J. Comparative proteome analysis of robust Saccharomyces cerevisiae insights into industrial continuous and batch fermentation. Applied Microbiology and Biotechnology, 2008, 81(2): 327–338

[26] Folch J, Lees M, Stanley G H S. A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry, 2008, 226: 497–509

[27] Yang S, Qiao B, Lu S H, Yuan Y J. Comparative lipidomics analysis of cellular development and apoptosis in two Taxus cell lines. Biochimica et Biophysica Acta (BBA)-. Molecular and Cell Biology of Lipids, 2007, 1771(5): 600–612

[28] Valero E, Millan C, Ortega J M. Changes in the lipid composition of saccharomyces cerevisiae race capensis (G-1) during alcoholic fermentation and flor film formation. LWT-Food Science and Technology, 2002, 35(7): 593–599

[29] Yokoyama K, Saitoh S, Ishida M, Yamakawa Y, Nakamura K, Inoue K, Taguchi R, Tokumura A, Nishijima M, Yanagida M, Setaka M. Very-long-chain fatty acid-containing PL accumulate in fatty acid synthase temperature-sensitive mutant strains of the fission yeast schizosaccharomyces pombe fas2/lsd1. Biochimica et Biophysica Acta, 2001, 1532(3): 223–233

[30] Schneiter R, Hitomi M, Ivessa A S, Fasch E V, Kohlwein S D, Tartakoff A M. A yeast acetyl-CoA carboxylase mutant links very long-chain fatty acid synthesis to structure and function of the nuclear membrane/pore complex. Molecular and Cellular Biology, 1996, 16(12): 7161–7172

[31] Schneiter R, Kohlwein S D. Organelle structure, function and inheritance in yeast: a role for fatty acid synthesis? Cell, 1997, 88(4): 431–434

[32] Stukey J E, Mcdonough V M, Martin C E. Isolation and characterization of OLE1, a gene affecting fatty acid desaturation from Saccharomyces cerevisiae. Journal of Biological Chemistry, 1989, 264(28): 16537–16544

[33] Mizoguchi H, Hara S. Ethanol-induced alterations in lipid composition of Saccharomyces cerevisiae in the presence of exogenous fatty acid. Journal of Fermentation and Bioengineering, 1997, 83(1): 12–16

[34] Black P N, Dirusso C C. Yeast acyl-CoA synthetases at the crossroads of fatty acid metabolism and regulation. Biochimica et Biophysica Acta (BBA)-. Molecular and Cell Biology of Lipids, 2007, 1771(3): 286–298

[35] Ishii I, Fukushima N, Ye X Q, Chun J. Lysophospholipid receptors: signaling and biology. Annual Review of Biochemistry, 2004, 73(1): 321–354

[36] Hoffmann P R, deCathelineau A M, Ogden C A, Leverrier Y, Bratton D L, Daleke D L, Ridley A J, Fadok V A, Henson P M. Phosphatidylserine (PS) induces PS receptormediated macropinocytosis and promotes clearance of apoptotic cells. Journal of Cell Biology, 2001, 155(4): 649–660

[37] Matsko C M, Hunter O C, Rabinowich H, Lotze M T, Amoscato A A. Mitochondrial lipid alterations during fas- and radiationinduced apoptosis. Biochemical and Biophysical Research Communications, 2001, 287(5): 1112–1120

[38] Qiu Y P, Cai G X, Su M M, Chen T L, Zheng X J, Xu Y, Ni Y, Zhao A H, Xu L X, Cai S J, Jia W. Serum metabolite profiling of human colorectal cancer using GC-TOFMS and UPLC-QTOFMS. Journal of Proteome Research, 2009, 8(10): 4844–4850

[39] Huo T G, Cai S, Lu X M, Sha Y, Yu M Y, Li F M. Metabonomic study of biochemical changes in the serum of type 2 diabetes mellitus patients after the treatment of metformin hydrochloride. Journal of Pharmaceutical and Biomedical Analysis, 2009, 49(4): 976–982

[40] Lewis B A, Engelman D M. Lipid bilayer thickness varies linearly with acyl chain-length in fluid phosphatidylcholine vesicles. Journal of Molecular Biology, 1983, 166(2): 211–217

[41] Paula S, Volkov A G, VanHoek A N, Haines T H, Deamer D W. Permeation of protons, potassium ions, and small polar molecules through phospholipid bilayers as a function of membrane thickness. Biophysical Journal, 1996, 70(1): 339–348

[42] Holub B J. Metabolism and function of myo-inositol and inositol glycerophospholipids. Annual Review of Nutrition, 1986, 6(1): 563–597

[43] Greenberg M L, Hubbell S, Lam C. Inositol regulates phosphatidylglycerolphosphate synthase expression in Saccharomyces cerevisiae. Molecular and Cellular Biology, 1988, 8(11): 4773–4779

Acknowledgments

The authors are grateful for the financial support from the National High-tech R&D Program (Grant No. 2012AA021204), the National Basic Research Program of China (Grant No. 2011CBA00802), and the National Natural Science Foundation of China (Grant No. 20736006 and Grant No. 21020102 040).

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg