文献类型：期刊文献

英文题名：Strategy of employing plug-and-play vectors and LC-MS screening to facilitate the discovery of natural products using Aspergillus oryzae

作者：Shi, Hanliang[1];Lin, Beibei[1];Zheng, Mengmeng[2,3,4];Gan, Fengyu[1];Lin, Zhi[2,3,4];Xin, Xiujuan[1];Zhao, Jian[1,5];Qu, Xudong[2,3,4];An, Faliang[1,6]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[2]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China;[3]Wuhan Univ, Sch Pharmaceut Sci, Key Lab Combinatorial Biosynth & Drug Discovery, Minist Educ, Wuhan 430071, Peoples R China;[4]Shanghai Jiao Tong Univ, Zhangjiang Inst Adv Study, Shanghai 201203, Peoples R China;[5]East China Univ Sci & Technol, Dept Appl Biol, 130 Mei Long Rd, Shanghai 200237, Peoples R China;[6]Marine Biomed Sci & Technol Innovat, 4 Lane 218,Haiji Sixth Rd,Platform Lin Gang Specia, Shanghai 201306, Peoples R China

年份：2025

卷号：12

期号：1

外文期刊名：BIORESOURCES AND BIOPROCESSING

收录：;EI(收录号：20250817910823);WOS:【SCI-EXPANDED(收录号:WOS:001392372100001)】；

基金：We sincerely acknowledge Prof. Ikuro Abe and Prof. Dan Hu for the kind provision of A. oryzae NSAR1 and the six original vectors, Prof. Mancheng Tang at Shanghai Jiao Tong University for the kind provision of Saccharomyces cerevisiae BJ5464-NpgA and pXW55, and Prof. Renxiang Tan at Nanjing University for the kind provision of Talaromyces sp. YE3016.

语种：英文

外文关键词：Plug-and-play vectors; LC-MS detection; Aspergillus oryzae; Heterologous expression; PamyB

摘要：Aspergillus oryzae is a widely used host for heterologous expression of fungal natural products. However, the vectors previously developed are not convenient for use and screening positive transformants by PCR and fermentation is time- and effort-consuming. Hence, three plug-and-play vectors were developed here for multi-gene expression and liquid chromatography mass spectrometry detection was introduced to screen positive transformants. Using rug BGC for verification, we demonstrated that the vectors we developed perform well and liquid chromatography mass spectrometry detection is feasible to screen positive transformants. For deleterious gene expression, PxyrA rather than PamyB was employed. Utilizing the toolkit described here to express natural products, dozen days can be saved.