文献类型：期刊文献

英文题名：Improved AP-3 production through combined ARTP mutagenesis, fermentation optimization, and subsequent genome shuffling

作者：Li, Juan[1];Guo, Siyu[1];Hua, Qiang[1];Hu, Fengxian[1,2]

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 Technol, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2021

卷号：43

期号：6

起止页码：1143

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20211310149599);WOS:【SCI-EXPANDED(收录号:WOS:000631311400001)】；

基金：This work was financially supported by the National Natural Science Foundation of China (21406070).

语种：英文

外文关键词：Actinosynnema pretiosum; Ansamitocin P-3; ARTP mutation; Genome shuffling

摘要：Ansamitocin (AP-3) is an ansamycins antibiotic isolated from Actinosynnema pretiosum and demonstrating high anti-tumor activity. To improve AP-3 production, the A. pretiosum ATCC 31565 strain was treated with atmospheric and room temperature plasma (ARTP). Four stable mutants were obtained by ARTP, of which the A. pretiosum L-40 mutant produced 242.9 mg/L AP-3, representing a 22.5% increase compared to the original wild type strain. With seed medium optimization, AP-3 production of mutant L-40 reached 307.8 mg/L; qRT-PCR analysis revealed that AP-3 biosynthesis-related gene expression was significantly up-regulated under optimized conditions. To further improve the AP-3 production, genome shuffling (GS) technology was used on the four A. pretiosum mutants by ARTP. After three rounds of GS combined with high-throughput screening, the genetically stable recombinant strain G3-96 was obtained. The production of AP-3 in the G3-96 strain was 410.1 mg/L in shake flask cultures, which was 44.5% higher than the L-40 production from the parental strain, and AP-3 was increased by 93.8% compared to the wild-type A. pretiosum. These results suggest that the combination of mutagenesis, seed medium optimization, and GS technology can effectively improve the AP-3 production capacity of A. pretiosum and provide an enabling methodology for AP-3 industrial production.