Abstract
Saccharomyces cerevisiae is a microorganism that is widely used for the bioproduction of useful substances, including ethanol. However, during bioproduction, yeast cells are subjected to various stresses. In particular, bioproduction via fermentation generates heat, leading to decreased rates of cell growth and fermentation. Improving the thermotolerance of yeast can help to maintain metabolic activity and bioproduction efficiency at high temperatures. In this chapter, we describe the improvement of thermotolerance in yeast through stepwise adaptive evolution under heat stress. The adaptation strategy, in which the cells were selected under two selective pressures (heat stress and growth rate), improved thermotolerance while maintaining growth rate. Through this adaptation strategy, a thermotolerant yeast strain, YK60-1, was successfully isolated after adaptation to 38 °C. Transcriptome and non-targeted metabolome analyses revealed that YK60-1 induced stress-responsive genes and accumulated more trehalose than the wild-type parent strain. Furthermore, comparative genomic analysis of the intermediate populations after adaptation to each elevated temperature revealed key mutations for improving thermotolerance in the CDC25 gene. A thermotolerant yeast strain was also reconstructed by introducing CDC25 mutations in the wild-type strain. CDC25 mutation is thought to alter global transcriptional regulation through downregulation of the cAMP/PKA pathway, leading to improved stress tolerance.
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References
Abe H, Fujita Y, Takaoka Y, Kurita E, Yano S, Tanaka N, Nakayama K (2009) Ethanol-tolerant Saccharomyces cerevisiae strains isolated under selective conditions by over-expression of a proofreading-deficient DNA polymerase δ. J Biosci Bioeng 108(3):199–204. https://doi.org/10.1016/j.jbiosc.2009.03.019
Almario MP, Reyes LH, Kao KC (2013) Evolutionary engineering of Saccharomyces cerevisiae for enhanced tolerance to hydrolysates of lignocellulosic biomass. Biotechnol Bioeng 110(10):2616–2623. https://doi.org/10.1002/bit.24938
Alper H, Moxley J, Nevoigt E, Fink GR, Stephanopoulos G (2006) Engineering yeast transcription machinery for improved ethanol tolerance and production. Science 314(5805):1565–1568. https://doi.org/10.1126/science.1131969
Boy-Marcotte E, Perrot M, Bussereau F, Boucherie H, Jacquet M (1998) Msn2p and Msn4p control a large number of genes induced at the diauxic transition which are repressed by cyclic AMP in Saccharomyces cerevisiae. J Bacteriol 180(5):1044–1052
Bro C, Nielsen J (2004) Impact of ‘ome’ analyses on inverse metabolic engineering. Metab Eng 6(3):204–211. https://doi.org/10.1016/j.ymben.2003.11.005
Broek D, Toda T, Michaeli T, Levin L, Birchmeier C, Zoller M, Powers S, Wigler M (1987) The S. cerevisiae CDC25 gene product regulates the RAS/adenylate cyclase pathway. Cell 48(5):789–799
Cakar ZP, Seker UO, Tamerler C, Sonderegger M, Sauer U (2005) Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae. FEMS Yeast Res 5(6–7):569–578. https://doi.org/10.1016/j.femsyr.2004.10.010
Cakar ZP, Alkim C, Turanli B, Tokman N, Akman S, Sarikaya M, Tamerler C, Benbadis L, Francois JM (2009) Isolation of cobalt hyper-resistant mutants of Saccharomyces cerevisiae by in vivo evolutionary engineering approach. J Biotechnol 143(2):130–138. https://doi.org/10.1016/j.jbiotec.2009.06.024
Cakar ZP, Turanli-Yildiz B, Alkim C, Yilmaz U (2012) Evolutionary engineering of Saccharomyces cerevisiae for improved industrially important properties. FEMS Yeast Res 12(2):171–182. https://doi.org/10.1111/j.1567-1364.2011.00775.x
Chen J, Pederson DS (1993) A distal heat shock element promotes the rapid response to heat shock of the HSP26 gene in the yeast Saccharomyces cerevisiae. J Biol Chem 268(10):7442–7448
Dragosits M, Mattanovich D (2013) Adaptive laboratory evolution—principles and applications for biotechnology. Microb Cell Fact 12:64. https://doi.org/10.1186/1475-2859-12-64
Eastmond DL, Nelson HC (2006) Genome-wide analysis reveals new roles for the activation domains of the Saccharomyces cerevisiae heat shock transcription factor (Hsf1) during the transient heat shock response. J Biol Chem 281(43):32909–32921. https://doi.org/10.1074/jbc.M602454200
Folch-Mallol JL, Martinez LM, Casas SJ, Yang R, Martinez-Anaya C, Lopez L, Hernandez A, Nieto-Sotelo J (2004) New roles for CDC25 in growth control, galactose regulation and cellular differentiation in Saccharomyces cerevisiae. Microbiology 150(9):2865–2879. https://doi.org/10.1099/mic.0.27144-0
Giardina C, Lis JT (1995) Dynamic protein-DNA architecture of a yeast heat shock promoter. Mol Cell Biol 15(5):2737–2744
Gibney PA, Lu C, Caudy AA, Hess DC, Botstein D (2013) Yeast metabolic and signaling genes are required for heat-shock survival and have little overlap with the heat-induced genes. Proc Natl Acad Sci USA 110(46):E4393–4402. https://doi.org/10.1073/pnas.1318100110
Gorner W, Durchschlag E, Martinez-Pastor MT, Estruch F, Ammerer G, Hamilton B, Ruis H, Schuller C (1998) Nuclear localization of the C2H2 zinc finger protein Msn2p is regulated by stress and protein kinase A activity. Genes Dev 12(4):586–597
Gorsich SW, Dien BS, Nichols NN, Slininger PJ, Liu ZL, Skory CD (2006) Tolerance to furfural-induced stress is associated with pentose phosphate pathway genes ZWF1, GND1, RPE1, and TKL1 in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 71(3):339–349. https://doi.org/10.1007/s00253-005-0142-3
Hahn JS, Hu Z, Thiele DJ, Iyer VR (2004) Genome-wide analysis of the biology of stress responses through heat shock transcription factor. Mol Cell Biol 24(12):5249–5256. https://doi.org/10.1128/MCB.24.12.5249-5256.2004
Kida K, Kume K, Morimura S, Sonoda Y (1992) Repeated-batch fermentation process using a thermotolerant flocculating yeast constructed by protoplast fusion. J Ferment Bioeng 74(3):169–173. https://doi.org/10.1016/0922-338X(92)90078-9
Kuroda K, Ueda M (2013) Arming technology in yeast-novel strategy for whole-cell biocatalyst and protein engineering. Biomolecules 3(3):632–650. https://doi.org/10.3390/biom3030632
Kuroda K, Ueda M (2017) Engineering of global regulators and cell surface properties toward enhancing stress tolerance in Saccharomyces cerevisiae. J Biosci Bioeng 124(6):599–605. https://doi.org/10.1016/j.jbiosc.2017.06.010
Kuyper M, Toirkens MJ, Diderich JA, Winkler AA, van Dijken JP, Pronk JT (2005) Evolutionary engineering of mixed-sugar utilization by a xylose-fermenting Saccharomyces cerevisiae strain. FEMS Yeast Res 5(10):925–934. https://doi.org/10.1016/j.femsyr.2005.04.004
Martinez-Pastor MT, Marchler G, Schuller C, Marchler-Bauer A, Ruis H, Estruch F (1996) The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). EMBO J 15(9):2227–2235
Matsui K, Kuroda K, Ueda M (2009) Creation of a novel peptide endowing yeasts with acid tolerance using yeast cell-surface engineering. Appl Microbiol Biotechnol 82(1):105–113. https://doi.org/10.1007/s00253-008-1761-2
Matsumoto K, Uno I, Oshima Y, Ishikawa T (1982) Isolation and characterization of yeast mutants deficient in adenylate cyclase and cAMP-dependent protein kinase. Proc Natl Acad Sci USA 79(7):2355–2359
Mitsuzawa H, Uno I, Oshima T, Ishikawa T (1989) Isolation and characterization of temperature-sensitive mutations in the RAS2 and CYR1 genes of Saccharomyces cerevisiae. Genetics 123(4):739–748
Pacheco A, Pereira C, Almeida MJ, Sousa MJ (2009) Small heat-shock protein Hsp12 contributes to yeast tolerance to freezing stress. Microbiology 155(6):2021–2028. https://doi.org/10.1099/mic.0.025981-0
Sakurai H, Takemori Y (2007) Interaction between heat shock transcription factors (HSFs) and divergent binding sequences: binding specificities of yeast HSFs and human HSF1. J Biol Chem 282(18):13334–13341. https://doi.org/10.1074/jbc.M611801200
Satomura A, Katsuyama Y, Miura N, Kuroda K, Tomio A, Bamba T, Fukusaki E, Ueda M (2013) Acquisition of thermotolerant yeast Saccharomyces cerevisiae by breeding via stepwise adaptation. Biotechnol Prog 29(5):1116–1123. https://doi.org/10.1002/btpr.1754
Satomura A, Kuroda K, Ueda M (2014) Environmental stress tolerance engineering by modification of cell surface and transcription factor in Saccharomyces cerevisiae. Curr Environ Eng 1(3):149–156. https://doi.org/10.2174/221271780103150522154913
Satomura A, Miura N, Kuroda K, Ueda M (2016) Reconstruction of thermotolerant yeast by one-point mutation identified through whole-genome analyses of adaptively-evolved strains. Sci Rep 6:23157. https://doi.org/10.1038/srep23157
Schmitt AP, McEntee K (1996) Msn2p, a zinc finger DNA-binding protein, is the transcriptional activator of the multistress response in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 93(12):5777–5782
Shi DJ, Wang CL, Wang KM (2009) Genome shuffling to improve thermotolerance, ethanol tolerance and ethanol productivity of Saccharomyces cerevisiae. J Ind Microbiol Biotechnol 36(1):139–147. https://doi.org/10.1007/s10295-008-0481-z
Thevelein JM, de Winde JH (1999) Novel sensing mechanisms and targets for the cAMP-protein kinase A pathway in the yeast Saccharomyces cerevisiae. Mol Microbiol 33(5):904–918
Ueda M, Tanaka A (2000) Genetic immobilization of proteins on the yeast cell surface. Biotechnol Adv 18(2):121–140
Wallace-Salinas V, Gorwa-Grauslund MF (2013) Adaptive evolution of an industrial strain of Saccharomyces cerevisiae for combined tolerance to inhibitors and temperature. Biotechnol Biofuels 6(1):151. https://doi.org/10.1186/1754-6834-6-151
Winkler JD, Kao KC (2014) Recent advances in the evolutionary engineering of industrial biocatalysts. Genomics 104(6):406-411. https://doi.org/10.1016/j.ygeno.2014.09.006
Wisselink HW, Toirkens MJ, Wu Q, Pronk JT, van Maris AJ (2009) Novel evolutionary engineering approach for accelerated utilization of glucose, xylose, and arabinose mixtures by engineered Saccharomyces cerevisiae strains. Appl Environ Microbiol 75(4):907–914. https://doi.org/10.1128/AEM.02268-08
Yazawa H, Iwahashi H, Uemura H (2007) Disruption of URA7 and GAL6 improves the ethanol tolerance and fermentation capacity of Saccharomyces cerevisiae. Yeast 24(7):551–560. https://doi.org/10.1002/yea.1492
Zou W, Ueda M, Yamanaka H, Tanaka A (2001) Construction of a combinatorial protein library displayed on yeast cell surface using DNA random priming method. J Biosci Bioeng 92(4):393–396
Zou W, Ueda M, Tanaka A (2002) Screening of a molecule endowing Saccharomyces cerevisiae with n-nonane-tolerance from a combinatorial random protein library. Appl Microbiol Biotechnol 58(6):806–812. https://doi.org/10.1007/s00253-002-0961-4
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Kuroda, K., Ueda, M. (2018). Adaptive Evolution of Yeast Under Heat Stress and Genetic Reconstruction to Generate Thermotolerant Yeast. In: Pontarotti, P. (eds) Origin and Evolution of Biodiversity. Springer, Cham. https://doi.org/10.1007/978-3-319-95954-2_2
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DOI: https://doi.org/10.1007/978-3-319-95954-2_2
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