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首页> 外文期刊>Journal of Biotechnology >Identification of gene targets eliciting improved alcohol tolerance in Saccharomyces cerevisiae through inverse metabolic engineering
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Identification of gene targets eliciting improved alcohol tolerance in Saccharomyces cerevisiae through inverse metabolic engineering

机译：通过逆向代谢工程鉴定引起酿酒酵母中耐酒精性提高的基因靶标

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The economic production of biofuels from renewable biomass using Saccharomyces cerevisiae requires tolerance to high concentrations of sugar and alcohol. Here we applied an inverse metabolic engineering approach to identify endogenous gene targets conferring improved alcohol tolerance in S. cerevisiae. After transformation with a S. cerevisiae genomic library, enrichment of the transformants exhibiting improved tolerance was performed by serial subculture in the presence of iso-butanol (1%). Through sequence analysis of the isolated plasmids from the selected transformants, four endogenous S. cerevisiae genes were identified as overexpression targets eliciting improved tolerance to both iso-butanol and ethanol. Overexpression of INO1, DOG1, HAL1 or a truncated form of MSN2 resulted in remarkably increased tolerance to high concentrations of iso-butanol and ethanol. Overexpression of INO1 elicited the highest ethanol tolerance, resulting in higher titers and volumetric productivities in the fermentation experiments performed with high glucose concentrations. In addition, the INO1-overexpressing strain showed a threefold increase in the specific growth rate as compared to that of the control strain under conditions of high levels of glucose (10%) and ethanol (5%). Although alcohol tolerance in yeast is a complex trait affected by simultaneous interactions of many genes, our results using a genomic library reveal potential target genes for better understanding and possible engineering of metabolic pathways underlying alcohol tolerance phenotypes
机译：使用酿酒酵母由可再生生物质经济生产生物燃料需要耐受高浓度的糖和酒精。在这里，我们应用逆代谢工程方法来鉴定赋予酿酒酵母耐酒精性的内源性基因靶标。用酿酒酵母基因组文库转化后，通过在异丁醇（1％）存在下的连续继代培养来富集表现出更高耐受性的转化子。通过对从选定的转化子中分离的质粒的序列分析，鉴定了四个内源啤酒酵母基因作为过表达靶标，其引起了对异丁醇和乙醇的改善的耐受性。 INO1，DOG1，HAL1或MSN2的截短形式的过表达导致对高浓度异丁醇和乙醇的耐受性显着提高。 INO1的过表达引起最高的乙醇耐受性，从而在高葡萄糖浓度的发酵实验中导致更高的滴度和体积生产率。另外，在高葡萄糖（10％）和乙醇（5％）的条件下，与对照菌株相比，INO1过表达菌株显示出比生长速率增加了三倍。尽管酵母中的酒精耐受性是受许多基因同时相互作用影响的复杂性状，但我们使用基因组文库的结果揭示了潜在的靶基因，可用于更好地理解和研究潜在的酒精耐受表型代谢途径

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