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首页> 外文期刊>Yeast >Gelatin expression from an engineered Saccharomyces cerevisiae CUP1 promoter in Pichia pastoris
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Gelatin expression from an engineered Saccharomyces cerevisiae CUP1 promoter in Pichia pastoris

机译：来自毕赤酵母的工程化酿酒酵母酿酒酵母Cup1启动子的明胶表达

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摘要

The methylotrophic yeast Pichia pastoris (reclassified as Komagataella phaffii) is a versatile protein expression system, yet many commonly used promoters have attributes undesirable for fermentation or its optimization. Hence, the copper-inducible CUP1 gene promoter from the related yeast Saccharomyces cerevisiae was used to express human gelatin. Multimerization of a potential copper response element in the CUP1 promoter, a S. cerevisiae Ace1p binding site, significantly increased gelatin expression. Expression was induced by copper in a dose-dependent fashion and was not dependent on cell density. Gelatin was additionally induced in standard copper-containing fermentation basal salts media. Removal of a S. cerevisiae heat shock factor (Hsf1p) binding site reduced copper-dependent gelatin induction suggesting that a similar protein may regulate this promoter in P. pastoris. This engineered copper inducible promoter expands the yeast recombinant protein production tool kit.
机译：甲基营养酵母毕赤酵母（重新分类为Komagatella phaffii）是一种多功能的蛋白质表达系统，但许多常用的启动子具有不适合发酵或优化的特性。因此，从相关的酿酒酵母中提取的铜诱导CUP1基因启动子被用于表达人类明胶。CUP1启动子（酿酒酵母Ace1p结合位点）中潜在铜反应元件的多聚化显著增加了明胶的表达。铜以剂量依赖性方式诱导表达，且不依赖于细胞密度。明胶在标准含铜发酵基础盐培养基中额外诱导。去除酿酒酵母热休克因子（Hsf1p）结合位点减少了铜依赖性明胶诱导，这表明类似的蛋白质可能在毕赤酵母中调节该启动子。这种工程铜诱导启动子扩展了酵母重组蛋白生产工具包。

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