为达到同步糖化发酵(simultaneous saccharification and fermentation,SSF)中菌株高温耐受能力提升的目的,该文以实验室保藏的马克斯克鲁维酵母(Kluyveromyces marxianus)GX-UN120为出发菌株,采用常压室温等离子体(atmospheric and room temperature plasma,ARTP)诱变技术,结合高温胁迫筛选获得1株具有较好耐高温能力和产乙醇能力的菌株GX-UN127。结果表明,诱变菌株GX-UN127在48 ℃培养72 h,OD600可达到1.27(原始菌株无法生长)。同时,当以100 g/L麸皮为原料,45 ℃ SSF 12 h,诱变菌株GX-UN127发酵乙醇的产量可达7.6 g/L,较出发菌株提升15.2%。在此基础上,结合生理指标和代谢组学进一步探究诱变菌株耐受高温的生理机制,研究表明半胱氨酸和蛋氨酸代谢、谷胱甘肽代谢、精氨酸和脯氨酸代谢、甘油磷脂代谢等代谢途径与诱变菌株GX-UN127的更强高温耐受性密切相关。此诱变选育耐高温马克斯克鲁维酵母菌株的方案可行,为该类菌株进行SSF生产燃料乙醇提供理论支持。
To enhance the high-temperature resistance of fermentation strains for simultaneous saccharification and fermentation, this study utilized Kluyveromyces marxianus GX-UN120, a strain known for its high ethanol yield, as the starting point.The atmospheric and room temperature plasma (ARTP) mutagenesis technology was employed to obtain a mutant strain, GX-UN127, which exhibited robust high-temperature resistance and ethanol production capabilities.This was achieved through a screening test for high-temperature resistance and subsequent rescreening for high-temperature growth ability.The findings of this study revealed that after 72 hours of cultivation at 48 ℃, the OD600 of GX-UN127 reached 1.27.Moreover, when using 100 g/L bran as the raw material, the ethanol yield reached 7.6 g/L after 12 h of simultaneous saccharification and fermentation at 45 ℃.This represented a 15.2% increase compared to the parent strain building upon these results, the study further investigated the physiological mechanism underlying the high-temperature tolerance of the mutant strain.By combining physiological indicators and metabolomics, it was determined that the metabolism of arginine and proline, as well as cysteine and methionine, were associated with the high-temperature tolerance of GX-UN127.Additionally, glycerophospholipid metabolism, glutathione metabolism, and other metabolic pathways were found to be closely related to the enhanced high-temperature tolerance observed in the mutant strain.Overall, this research demonstrates the feasibility of breeding high-temperature resistant Kluyveromyces marxianus strains through mutation.The findings provide valuable theoretical support for the simultaneous saccharification and fermentation of this strain for ethanol production.
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