Abstract
To improve S-Adenosyl-L-methionine (a compound with important physiological functions, SAM) production, atmospheric and room temperature plasma and ultraviolet-LiCl mutagenesis were carried out with Saccharomyces cerevisiae strain ZY 1–5. The mutants were screened with ethionine, L-methionine, nystatin and cordycepin as screening agents. Adaptive evolution of a positive mutant UV6-69 was further performed by droplet microfluidics cultivation with ethionine as screening pressure. After adaptation, mutant T11-1 was obtained. Its SAM titer in shake flask fermentation reached 1.31 g/L, which was 191% higher than that of strain ZY 1–5. Under optimal conditions, the SAM titer and biomass of mutant T11-1 in 5 L bioreactor reached 10.72 g/L and 105.9 g dcw/L (142.86% and 34.22% higher than those of strain ZY 1–5), respectively. Comparative transcriptome analysis between strain ZY 1–5 and mutant T11-1 revealed the enhancements in TCA cycle and gluconeogenesis/glycolysis pathways as well as the inhibitions in serine and ergosterol synthesis of mutant T11-1. The elevated SAM synthesis of mutant T11-1 may attribute to the above changes. Taken together, this study is helpful for industrial production of SAM.
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References
Beneyton T, Wijaya I, Postros P et al (2016) High-throughput screening of filamentous fungi using nanoliter-range droplet-based microfluidics. Sci Rep 6(1):1–10. https://doi.org/10.1038/srep27223
Cao X, Yang M, Xia Y et al (2012) Strain improvement for enhanced production of S-adenosyl-L-methionine in Saccharomyces cerevisiae based on ethionine-resistance and SAM synthetase activity. Ann Microbiol 62(4):1395–1402. https://doi.org/10.1007/s13213-011-0389-0
Chan SY, Appling DR (2003) Regulation of S-adenosylmethionine levels in Saccharomyces cerevisiae. J Biol Chem 278(44):43051–43059. https://doi.org/10.1074/jbc.M308696200
Chen YW (2018) Cofactor engineering strategy for enhanced S-adenosylmethionine production in Saccharomyces cerevisiae. Chin J Biotechnol 34(2):246–254. https://doi.org/10.13345/j.cjb.170174
Chen H, Wang Z, Cai H et al (2016a) Progress in the microbial production of S-adenosyl-L-methionine. World J Microbiol Biotechnol 32(9):1–8. https://doi.org/10.1007/s11274-016-2102-8
Chen H, Wang Z, Wang Z et al (2016b) Improving methionine and ATP availability by MET6 and SAM2 co-expression combined with sodium citrate feeding enhanced SAM accumulation in Saccharomyces cerevisiae. World J Microbiol Biotechnol 32(4):1–10. https://doi.org/10.1007/s11274-016-2010-y
Chen H, Zhu N, Wang Y et al (2021) Increasing glycolysis by deletion of kcs1 and arg82 improved S-adenosyl-L-methionine production in Saccharomyces cerevisiae. AMB Express 11(1):1–16. https://doi.org/10.1186/s13568-021-01179-8
Choi E, Park B, Lee S et al (2009) Increased production of S-adenosyl-L-methionine using recombinant Saccharomyces cerevisiae sake K6. Korean J Chem Eng 26(1):156–159. https://doi.org/10.1007/s11814-009-0025-x
Cregg JM, Hessler BK, A, et al (1985) Pichia pastoris as a host system for transformations. Mol Cell Biol 5(12):3376–3385. https://doi.org/10.1128/mcb.5.12.3376-3385.1985
Dong H, Liu F, Tan T (2006) Study on screening the strain yielding highly s-adenosylmethionine by complex mutation and its culture manner. Chin J Bioprocess Eng 4(2):20–23
Feng XX, Wang YQ (2011) Breeding of Saccharomyces cerevisiae giving an enhanced yield of S-adenosyl-L-methionine by composite mutagenesis and protoplast fusion. J Beijing Uni Chem Technol 38(6):87–92
Fontecave M, Atta M, Mulliez E (2004) S-adenosylmethionine: nothing goes to waste. Trends Biochem Sci 29(5):243–249. https://doi.org/10.1016/j.tibs.2004.03.007
Guo X, Wang L, Zhang C et al (2021) Technology development and instrumentation of a high-throughput and automated microbial microdroplet culture system for microbial evolution and screening. Chin J Biotechnol 37(3):991–1003. https://doi.org/10.13345/j.cjb.200667
Herrero P, Galindez J, Ruiz N et al (1995) Transcriptional regulation of the Saccharomyces cerevisiae HXK1, HXK2 and GLK1 genes. Yeast 11(2):137–144. https://doi.org/10.1002/yea.320110205
Huang Y, Gou X, Hu H et al (2012) Enhanced S-adenosyl-L-methionine production in Saccharomyces cerevisiae by spaceflight culture, overexpressing methionine adenosyltransferase and optimizing cultivation. J Appl Microbiol 112(4):683–694. https://doi.org/10.1111/j.1365-2672.2012.05251.x
Jian X, Guo X, Wang J et al (2020) Microbial microdroplet culture system (MMC): an integrated platform for automated, high-throughput microbial cultivation and adaptive evolution. Biotechnol Bioeng 117(6):1724–1737. https://doi.org/10.1002/bit.27327
Kanai M, Yasuda N, Morimoto T et al (2019) Breeding of a cordycepin-resistant and adenosine kinase-deficient sake yeast strain that accumulates high levels of S-adenosylmethionine. Biosci Biotechnol 83(8):1530–1537. https://doi.org/10.1080/09168451.2019.1571896
Kim SY, Choi JS, Park C et al (2010) Ethyl pyruvate stabilizes hypoxia-inducible factor 1 alpha via stimulation of the TCA cycle. Cancer Lett 295(2):236–241. https://doi.org/10.1016/j.canlet.2010.03.006
Lee SW, Park BS, Choi ES et al (2010) Overexpression of ethionine resistance gene for maximized production of S-adenosylmethionine in Saccharomyces cerevisiae sake kyokai No. 6. Korean J Chem Eng 27(2):587–589. https://doi.org/10.1007/s11814-010-0100-3
Leng N, Dawson JA, Thomson JA et al (2013) EBSeq: an empirical Bayes hierarchical model for inference in RNA-seq experiments. Bioinformatics 29(8):1035–1043. https://doi.org/10.1093/bioinformatics/btt087
Li G, Li H, Tan Y et al (2020) Improved S-adenosyl-l-methionine production in Saccharomyces cerevisiae using tofu yellow serofluid. J Biotechnol 309:100–106. https://doi.org/10.1016/j.jbiotec.2020.01.004
Lin JP, Tian J, You JF et al (2004) An effective strategy for the co-production of S-adenosyl-L-methionine and glutathione by fed-batch fermentation. Biochem Eng J 21(1):19–25. https://doi.org/10.1016/j.bej.2004.04.013
Liu W, Tang D, Shi R et al (2019) Efficient production of S-adenosyl-l-methionine from dl-methionine in metabolic engineered Saccharomyces cerevisiae. Biotechnol Bioeng 116(12):3312–3323. https://doi.org/10.1002/bit.27157
Matsushika A, Goshima T, Fujii T et al (2012) Characterization of non-oxidative transaldolase and transketolase enzymes in the pentose phosphate pathway with regard to xylose utilization by recombinant Saccharomyces cerevisiae. Enzyme Microb Tech 51(1):16–25. https://doi.org/10.1016/j.enzmictec.2012.03.008
Qin HB, Niu K, Wang YS (2020) Mutation breeding of S-adenosylmethionine-producing yeast. Food Ferment Ind 46(21):23–27. https://doi.org/10.13995/j.cnki.11-1802/ts.024211
Schlenk F, Zydek CR, Ehninger DJ et al (1965) The production of S-adenosyl-L-methionine and S-adenosyl-L-ethionine by yeast. Enzymologia 29(3):283–298
Shobayashi M, Mukai N, Iwashita K et al (2006) A new method for isolation of S-adenosylmethionine (SAM)-accumulating yeast. Appl Microbiol Biotechnol 69(6):704–710. https://doi.org/10.1007/s00253-005-0009-7
Trapnell C, Williams BA, Pertea G et al (2010) Transcript assembly and quantification by RNA-seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28(5):511–515. https://doi.org/10.1038/nbt.1621
Umekawa M, Hamada K, Isono N et al (2020) The Emi2 protein of Saccharomyces cerevisiae is a hexokinase expressed under glucose limitation. J Appl Microbiol 67(4):103–109. https://doi.org/10.5458/jag.jag.JAG-2020_0007
Wang L, Zhao H, He D et al (2020) Insights into the molecular-level effects of atmospheric and room-temperature plasma on mononucleotides and single-stranded homo-and hetero-oligonucleotides. Sci Rep 10(1):1–11. https://doi.org/10.1038/s41598-020-71152-1
Zhang X, Zhang XF, Li HP et al (2014) Atmospheric and room temperature plasma (ARTP) as a new powerful mutagenesis tool. Appl Microbiol Biotechnol 98(12):5387–5396. https://doi.org/10.1007/s00253-014-5755-y
Zhao W, Shi F, Hang B et al (2016) The improvement of SAM accumulation by integrating the endogenous methionine adenosyltransferase gene SAM2 in genome of the industrial Saccharomyces cerevisiae strain. Appl Biochem Biotechnol 178(6):1263–1272. https://doi.org/10.1007/s12010-015-1943-1
Zhao WJ, Huang L, Xu ZN (2017) Research progress on the synthesis of S-adenosyl-L-methionine in microorganism. Biol Bull 33(1):99–105. https://doi.org/10.13560/j.cnki.biotech.bull.1985.2017.01.010
Acknowledgements
This research is supported by National Key R&D Program of China (2020YFA0907900), Ministry of Science and Technology of the People´s Republic of China, and Zhejiang Provincial Natural Science Foundation (No. LY22C010005, LY17C010005).
Funding
Ministry of Science and Technology of the People’s Republic of China, 2020YFA0907900, Yuanshan Wang, Natural Science Foundation of Zhejiang Province,LY22C010005, Chunyue Weng, LY17C010005, Yuanshan Wang
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MZ, LM and QH, implementation of the experiment, preparation of manuscript. WY, QH, WC, LZ, English check. WY, WC, HZ, LZ and ZY supervision, experimental program design.
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Weng, C., Mi, Z., Li, M. et al. Improvement of S-adenosyl-L-methionine production in Saccharomyces cerevisiae by atmospheric and room temperature plasma-ultraviolet compound mutagenesis and droplet microfluidic adaptive evolution. 3 Biotech 12, 223 (2022). https://doi.org/10.1007/s13205-022-03297-x
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DOI: https://doi.org/10.1007/s13205-022-03297-x