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线粒体靶向增加异源缬氨酸同化途径在酿酒酵母中的比活性-外文翻译/外文文献翻译

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发布于 2017-1-28 23:19:11 | 显示全部楼层 |阅读模式
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外文标题: Mitochondrial targeting increases specific activity of a heterologous valine assimilation pathway in Saccharomyces cerevisiae
外文摘要: Bio-based isobutantol is a sustainable ‘drop in’ substitute for petroleum-based fuels. However, well- studied production routes, such as the Ehrlich pathway, have yet to be commercialized despite more than a century of research. The more versatile bacterial valine catabolism may be a competitive alternate route producing not only an isobutanol precursor but several carboxylic acids with applications as biomono- mers, and building blocks for other advanced biofuels. Here, we transfer the first two committed steps of the pathway from pathogenic Pseudomonas aeruginosa PAO1 to yeast to evaluate their activity in a safer model organism. Genes encoding the heteroligomeric branched chain keto-acid dehydrogenase (BCKAD; bkdA1, bkdA2, bkdB, lpdV), and the homooligomeric acyl-CoA dehydrogenase (ACD; acd1) were tagged with fluorescence epitopes and targeted for expression in either the mitochondria or cytoplasm of S. cerevisiae. We verified the localization of our constructs with confocal fluorescence microscopy before measuring the activity of tag-free constructs. Despite reduced heterologous expression of mitochondria- targeted enzymes, their specific activities were significantly improved with total enzyme activities up to 138% greater than those of enzymes expressed in the cytoplasm. In total, our results demonstrate that the choice of protein localization in yeast has significant impact on heterologous activity, and suggests a new path forward for isobutanol production.
英关键词: Pseudomonas Isobutanol Dehydrogenase Mitochondria Saccharomyces cerevisiae Metabolic engineering
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译文标题: 线粒体靶增加异源缬氨酸同化途径在酿酒酵母中的比活性
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译文摘要: 生物基异丁醇是石油基燃料的一种可持续的而且“随手可得”的替代品。然而,尽管花了一个多世纪的调查,精心研究的生产方式如艾利希途径仍未能商业化。更多功能的细菌缬氨酸分解代谢可能是具有竞争性的替代途径,不仅能产出异丁醇前体,还可得到几种羧酸,作为生物单体进行应用,并为其他高级生物燃料的生产奠定基石。本文从致病性铜绿假单胞菌PAO1到酵母改变了路径的前两个关键步骤,在更安全的模型生物体中评估活性。用荧光表位标记编码异寡聚体支链酮酸脱氢酶(BCKAD; bkdA1, bkdA2, bkdB, lpdV)和均低聚酰基-CoA脱氢酶的基因,并在线粒体或细胞质表达基因。测量无标记物的活性前,用共聚焦荧光显微镜确认目标的定位。尽管线粒体靶向酶的异源表达减弱,而与在细胞质中表达的酶相比,比活性显著提高,酶的总活性达到138%。总体上,结果表明,酵母中蛋白质定位对异源活性有显着影响,昭示了异丁醇生产的新路径。
译文格式: WORD
中关键词: 关键词:假单胞菌 异丁醇 脱氢酶 线粒体 酿酒酵母 代谢工程
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线粒体靶增加异源缬氨酸同化途径在酿酒酵母中的比活性.pdf

 
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