通过工程化酿酒酵母在较低 pH 条件下进行纤维二糖发酵的转运蛋白工程。
Transporter engineering for cellobiose fermentation under lower pH conditions by engineered Saccharomyces cerevisiae.
机构信息
Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
出版信息
Bioresour Technol. 2017 Dec;245(Pt B):1469-1475. doi: 10.1016/j.biortech.2017.05.138. Epub 2017 May 22.
The aim of this study was to engineer cellodextrin transporter 2 (CDT-2) from Neurospora crassa for improved cellobiose fermentation under lower pH conditions by Saccharomyces cerevisiae. Through directed evolution, a mutant CDT-2 capable of facilitating cellobiose fermentation under lower pH conditions was obtained. Specifically, a library of CDT-2 mutants with GFP fusion was screened by flow cytometry and then serial subcultured to isolate a CDT-2 mutant capable of transporting cellobiose under acidic conditions. The engineered S. cerevisiae expressing the isolated mutant CDT-2 (I96N/T487A) produced ethanol with a specific cellobiose consumption rate of 0.069g/gcell/h, which was 51% and 55% higher than those of the strains harboring wild-type CDT-1 and CDT-2 in a minimal medium with 2g/L of acetic acid.
本研究旨在通过酿酒酵母对来自粗糙脉孢菌的纤维二糖转运蛋白 2（CDT-2）进行工程改造，使其在较低 pH 条件下提高纤维二糖发酵能力。通过定向进化，获得了一种能够在较低 pH 条件下促进纤维二糖发酵的突变型 CDT-2。具体来说，通过流式细胞术筛选带有 GFP 融合的 CDT-2 突变文库，然后进行连续传代培养，以分离出一种能够在酸性条件下转运纤维二糖的 CDT-2 突变体。表达分离的突变型 CDT-2（I96N/T487A）的工程化酿酒酵母产生乙醇的比细胞内纤维二糖消耗率为 0.069g/gcell/h，比含有野生型 CDT-1 和 CDT-2 的菌株在含 2g/L 乙酸的最小培养基中分别提高了 51%和 55%。
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