Enhanced Protein Production by Sorbitol Co-feeding with Methanol in Recombinant Pichia pastoris Strains

文献类型： 外文期刊

第一作者： Chen, Li

作者： Chen, Li;Liu, Yamei;Chen, Li;Mohsin, Ali;Chu, Ju;Zhuang, Yingping;Guo, Meijin;Liu, Yamei

作者机构：

关键词： sorbitol;co-feeding;multi-copy;Pichia pastoris;porcine insulin precursor

期刊名称：BIOTECHNOLOGY AND BIOPROCESS ENGINEERING （ 影响因子：2.836； 五年影响因子：2.281 ）

ISSN： 1226-8372

年卷期： 2017 年 22 卷 6 期

页码：

收录情况： SCI

摘要： Pichia pastoris strains carrying 1, 6, 12, and 18 copies of the porcine insulin precursor (PIP) gene, were employed to investigate the effects of sorbitol co-feeding with methanol on the physiology of the strains. Multicopy clones of the methylotrophic yeast were generated to vary the PIP gene dosage and recombinant proteins. Elevated gene dosage increased levels of the recombinant PIP protein when methanol served as the sole carbon and energy source i.e., an increase of 1.9% for a strain carrying 1 copy, 42.6% for a strain carrying 6 copies, 34.7% for a strain carrying 12 copies and 80.9% for a strain carrying 18 copies, respectively (using sorbitol co-feeding with methanol during the induction phase). However, it had no significant influence on a lower gene dosage strain (1 copy), but this approach affirmed enhancement in cell growth and PIP production for higher gene dosage strain (6, 12, and 18 copies) via using sorbitol co-feeding with methanol. Additionally, the co-feeding strategy could hold vital importance for recombinant protein production by a multi-copy P. pastoris system.

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