Collagen and Gelatin Molecular Biology, FibroGen, Inc., San Francisco, CA, USA.
Yeast. 2021 Jun;38(6):382-387. doi: 10.1002/yea.3554. Epub 2021 Mar 15.
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.
甲醇营养型酵母巴斯德毕赤酵母（已重分类为毕赤酵母）是一种多功能的蛋白表达系统，但许多常用的启动子具有不适合发酵或优化的特性。因此，使用相关酵母酿酒酵母中的铜诱导 CUP1 基因启动子来表达人明胶。在 CUP1 启动子中潜在铜反应元件的多聚化，即酿酒酵母 Ace1p 结合位点，显著增加了明胶的表达。表达以依赖于铜剂量的方式诱导，而不依赖于细胞密度。在标准含铜发酵基础盐培养基中也可以诱导明胶。去除酿酒酵母热休克因子（Hsf1p）结合位点会降低铜依赖性明胶诱导，这表明类似的蛋白可能在巴斯德毕赤酵母中调节该启动子。这个经过工程设计的铜诱导启动子扩展了酵母重组蛋白生产工具包。
