YPRO15C过表达对酿酒酵母BY4742对糠醛抑制剂抗性的贡献

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2023-06-01 DOI:10.33073/pjm-2023-019
Getachew Tafere Abrha, Qian Li, Xiaolin Kuang, Difan Xiao, Ellen Ayepa, Jinjian Wu, Huan Chen, Zhengyue Zhang, Yina Liu, Xiumei Yu, Quanju Xiang, Menggen Ma
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引用次数: 1

摘要

木质纤维素生物质仍然被认为是生物乙醇生产的可行来源。酿酒酵母可以适应解毒木质纤维素来源的抑制剂,包括糠醛。菌株性能的耐受性已经通过在糠醛抑制剂挑战后细胞增殖的滞后期的程度来测量。本工作的目的是通过体内同源重组法过表达YPR015C,获得一株对糠醛耐受的酵母菌。对过表达酵母菌的生理观察表明,其对糠醛的抗性比亲本菌株更强。荧光显微镜显示,与亲本菌株相比,由于糠醛抑制剂的有害作用,酶还原酶活性和氧活性物质的积累有所提高。比较转录组学分析发现,在滞后期生长对糠醛的应激反应中,YPR015C过表达菌株可能涉及氨基酸生物合成、氧化应激、细胞壁反应、热休克蛋白和线粒体相关蛋白等79个基因。对酵母生长滞后期生存和适应糠醛胁迫的耐受性进行了时间过程研究,结果表明,酵母对糠醛胁迫的耐受性涉及多种功能范畴的上调和下调基因。本研究全面扩大了我们对YPR015C过表达菌株对糠醛胁迫耐受的生理和分子机制的认识。重组质粒构建示意图。a) pUG6-TEF1p-YPR015C, b)重组质粒pUG6-TEF1p-YPR与酿酒酵母染色体DNA的整合图。
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Contribution of YPRO15C Overexpression to the Resistance of Saccharomyces cerevisiae BY4742 Strain to Furfural Inhibitor.

Lignocellulosic biomass is still considered a feasible source of bioethanol production. Saccharomyces cerevisiae can adapt to detoxify lignocellulose-derived inhibitors, including furfural. Tolerance of strain performance has been measured by the extent of the lag phase for cell proliferation following the furfural inhibitor challenge. The purpose of this work was to obtain a tolerant yeast strain against furfural through overexpression of YPR015C using the in vivo homologous recombination method. The physiological observation of the overexpressing yeast strain showed that it was more resistant to furfural than its parental strain. Fluorescence microscopy revealed improved enzyme reductase activity and accumulation of oxygen reactive species due to the harmful effects of furfural inhibitor in contrast to its parental strain. Comparative transcriptomic analysis revealed 79 genes potentially involved in amino acid biosynthesis, oxidative stress, cell wall response, heat shock protein, and mitochondrial-associated protein for the YPR015C overexpressing strain associated with stress responses to furfural at the late stage of lag phase growth. Both up- and down-regulated genes involved in diversified functional categories were accountable for tolerance in yeast to survive and adapt to the furfural stress in a time course study during the lag phase growth. This study enlarges our perceptions comprehensively about the physiological and molecular mechanisms implicated in the YPR015C overexpressing strain's tolerance under furfural stress. Construction illustration of the recombinant plasmid. a) pUG6-TEF1p-YPR015C, b) integration diagram of the recombinant plasmid pUG6-TEF1p-YPR into the chromosomal DNA of Saccharomyces cerevisiae.

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ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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