文献类型：期刊文献

英文题名：Methylotrophic yeast Pichia pastoris as a chassis organism for polyketide synthesis via the full citrinin biosynthetic pathway

作者：Xue, Ying[1];Kong, Chuixing[1];Shen, Wei[1];Bai, Chenxiao[1];Ren, Yanna[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：2017

卷号：242

起止页码：64

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20165203160600);WOS:【SCI-EXPANDED(收录号:WOS:000392772500006)】；

基金：We thank Prof. Russell J. Cox, Leibniz Universitat Hannover, for the direction of citrinin gene cluster mining. This work was funded by the National Natural Science Funds of China (41306121) and Fundamental Research Funds for the Central Universities of China (22A201514040), and partially supported by Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Polyketides; Pichia pastoris; Heterologous biosynthesis; Citrinin; Biosynthetic pathway

摘要：With the rapid development of synthetic biology, exploring various chassis organisms has become necessary to improve the heterologous biosynthesis of natural products and pharmaceuticals. In this study, we tested the potential of the industrial methylotrophic yeast strain Pichia pastoris for the heterologous synthesis of polyketides. A recombinant P. pastoris GS-pksCT-npgA carrying the Monascus purpureus citrinin polyketide synthase gene pksCT and the Aspergillus nidulans phosphopantetheinyl transferase gene npgA was constructed. Subsequently, a specific compound was isolated and identified as citrinin intermediate trimethylated pentaketide aldehyde. On account of the hypothetic functions of the genes in the citrinin gene cluster, mpl1 encoding serine hydrolase, mpl2 encoding oxygenase, and mpl4 encoding dehydrogenase were gradually expressed. Proteins were also normally expressed, but a new compound was undetected. Basing on the recently reported citrinin gene cluster in Monascus ruber, we obtained two other genes (mpl6 and mpl7) participating in citrinin biosynthesis by genome walking in M. purpureus. Then, we co-transformed intron-removed mpl6 and mpl7 into the P. pastoris strain carrying pksCT, npgA, mpl1, mpl2, and mpl4. All genes were activated by the methanol-induced AOX1 promoter, and a complete biosynthetic pathway of citrinin was assembled. Finally, citrinin was successfully produced under methanol induction in P. pastoris. These results prove that P. pastoris is a promising chassis organism for polyketide production. (C) 2016 Elsevier B.V. All rights reserved.