文献类型：期刊文献

英文题名：Engineered monoculture and co-culture of methylotrophic yeast for de novo production of monacolin J and lovastatin from methanol

作者：Liu, Yiqi[1];Tu, Xiaohu[1];Xu, Qin[1];Bai, Chenxiao[1];Kong, Chuixing[1];Liu, Qi[1];Yu, Jiahui[1];Peng, Qiangqiang[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, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2018

卷号：45

起止页码：189

外文期刊名：METABOLIC ENGINEERING

收录：;EI(收录号：20180804824666);WOS:【SCI-EXPANDED(收录号:WOS:000424292100020)】；

基金：This work was supported Fundamental Research Funds for the Shanghai Science and Technology Innovation Action Plan (17JC1402400), Fundamental Research Funds for the Central Universities (22A201514040) and Talent Program of School of Biotechnology in East China University of Science and Technology. The authors would like to thank Prof. John C. Vederas (University of Alberta) for kindly providing the standard of monacolin J.

语种：英文

外文关键词：Methanol; Methylotrophic yeast; Co-culture; Heterologous biosynthesis; Monacolin J; Lovastatin

摘要：As a promising one-carbon renewable substrate for industrial biotechnology, methanol has attracted much attention. However, engineering of microorganisms for industrial production of pharmaceuticals using a methanol substrate is still in infancy. In this study, the methylotrophic yeast Pichia pastoris was used to produce anti-hypercholesterolemia pharmaceuticals, lovastatin and its precursor monacolin J, from methanol. The biosynthetic pathways for monacolin J and lovastatin were first assembled and optimized in single strains using single copies of the relevant biosynthetic genes, and yields of 60.0 mg/L monacolin J and 14.4 mg/L lovastatin were obtained using methanol following pH controlled monoculture. To overcome limitations imposed by accumulation of intermediates and metabolic stress in monoculture, approaches using pathway splitting and co-culture were developed. Two pathway splitting strategies for monacolin J, and four for lovastatin were tested at different metabolic nodes. Biosynthesis of monacolin J and lovastatin was improved by 55% and 71%, respectively, when the upstream and downstream modules were separately accommodated in two different fluorescent strains, split at the metabolic node of dihydromonacolin L. However, pathway distribution at monacolin J blocked lovastatin biosynthesis in all designs, mainly due to its limited ability of crossing cellular membranes. Bioreactor fermentations were tested for the optimal co-culture strategies, and yields of 593.9 mg/L monacolin J and 250.8 mg/L lovastatin were achieved. This study provides an alternative method for production of monacolin J and lovastatin and reveals the potential of a methylotrophic yeast to produce complicated pharmaceuticals from methanol.