文献类型：期刊文献

英文题名：Improved methanol-derived lovastatin production through enhancement of the biosynthetic pathway and intracellular lovastatin efflux in methylotrophic yeast

作者：Liu, Yiqi[1];Bai, Chenxiao[1];Xu, Qin[1];Yu, Jiahui[1];Zhou, Xiangshan[1];Zhang, Yuanxing[1,2];Cai, Menghao[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, Shanghai 200237, Peoples R China

年份：2018

卷号：5

期号：1

外文期刊名：BIORESOURCES AND BIOPROCESSING

收录：;EI(收录号：20224513075920);WOS:【SCI-EXPANDED(收录号:WOS:000461106800022)】；

基金：This work was supported by Fundamental Research Funds for the Shanghai Science and Technology Innovation Action Plan (Grant number 17JC1402400) and the Talent Program of the School of Biotechnology at East China University of Science and Technology.

语种：英文

外文关键词：Enhanced biosynthetic pathway; Lovastatin; Methanol; Pichia pastoris; Statin pump protein

摘要：Background: Methanol has attracted interest as a substrate for improvement of product titers and yields of bioprocesses, because methanol has a more reduced state than sugars do and can be renewably synthesized from abundant natural gas supplies. Our aim was to engineer methylotrophic Pichia pastoris to convert methanol into value-added lovastatin more productively. Results: A strengthened biosynthetic pathway of lovastatin was constructed through the assembly of three modules with increasing module-specific antibiotic stress in the methylotrophic yeast P pastoris. The resulting strain (P.p/LV_V#9) produced 287.5 +/- 2.0 mg/L lovastatin in a 5-L bioreactor from methanol. The production was further improved by identification and overexpression of a statin pump protein, TapA, a membrane protein capable of lovastatin efflux out of the cell. A TapA-overexpressing strain, P.p/LV_V#9-TapA, produced 419.0 +/- 9.5 mg/L lovastatin from methanol: 46% more than P.p/LV_V#9 did, and 520% more relative to the strain (P.p/LV_SC) with single-copy genes. Conclusions: A methylotrophic yeast strain producing 419.0 +/- 9.5 mg/L lovastatin was constructed by optimization of biosynthetic gene dosages and coexpression of a statin pump protein; these results proved that P. pastoris is a promising chassis organism for natural-product biosynthesis. A membrane protein,TapA was found to perform the function of exporting intracellular lovastatin and enhanced lovastatin production.