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Construction of a xylose-metabolizing yeast by genome integration of xylose isomerase gene and investigation of the effect of xylitol on fermentation

机译:木糖异构酶基因的基因组整合构建木糖代谢酵母并研究木糖醇对发酵的影响

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A yeast with the xylose isomerase (XI) pathway was constructed by the multicopy integration of XI overexpression cassettes into the genome of the Saccharomyces cerevisiae MT8-1 strain. The resulting yeast strain successfully produced ethanol from both xylose as the sole carbon source and a mixed sugar, consisting of xylose and glucose, without any adaptation procedure. Ethanol yields in the fermentation from xylose and mixed sugar were 61.9% and 62.2% of the theoretical carbon recovery, respectively. Knockout of GRE3, a gene encoding nonspecific aldose reductase, of the host yeast strain improved the fermentation profile. Not only specific ethanol production rates but also xylose consumption rates was improved more than twice that of xylose-metabolizing yeast with the XI pathway using GRE3 active yeast as the host strain. In addition, it was demonstrated that xylitol in the medium exhibits a concentration-dependent inhibition effect on the ethanol production from xylose with the yeast harboring the XI-based xylose metabolic pathway. From our findings, the combination of XI-pathway integration and GRE3 knockout could be result in a consolidated xylose assimilation pathway and increased ethanol productivity.
机译:通过将XI过表达盒多拷贝整合到酿酒酵母MT8-1菌株的基因组中,构建了具有木糖异构酶(XI)途径的酵母。所得的酵母菌株成功地从作为唯一碳源的木糖和由木糖和葡萄糖组成的混合糖中成功生产出乙醇,无需任何适应程序。木糖和混合糖在发酵中的乙醇收率分别为理论碳回收率的61.9%和62.2%。宿主酵母菌株的编码非特异性醛糖还原酶非编码基因GRE3的敲除改善了发酵特性。使用GRE3活性酵母作为宿主菌株,通过XI途径,不仅特定的乙醇生产速率提高,而且木糖消耗速率也提高了两倍以上。另外,已经证明,培养基中的木糖醇对木糖产生的乙醇具有浓度依赖性的抑制作用,而酵母具有基于XI的木糖代谢途径。根据我们的发现,XI途径整合和GRE3敲除的组合可能导致巩固的木糖同化途径并提高乙醇生产率。

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