文献类型：期刊文献

英文题名：Metabolomics analysis of Actinosynnema pretiosum with improved AP-3 production by enhancing UDP-glucose biosynthesis

作者：Liu, Ting[1];Jin, Ziwen[1];Wang, Ziwei[1];Chen, Jun[1];Wei, Liu-Jing[1];Hua, Qiang[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：2020

卷号：130

期号：1

起止页码：36

外文期刊名：JOURNAL OF BIOSCIENCE AND BIOENGINEERING

收录：;EI(收录号：20201108305785);WOS:【SCI-EXPANDED(收录号:WOS:000614230100006)】；

基金：This study was supported by the 111 Project (B18022), Research Program of State Key Laboratory of Bioreactor Engineering, China; Research Fund for the Doctoral Program of Higher Education of China (20130074110002), China. The authors declare no conflict of interest.

语种：英文

外文关键词：Actinosynnema pretiosum; Ansamitocin P-3; Metabolomics; Gene transcription; UDP-glucose supply

摘要：Ansamitocin P-3 (AP-3) shows strong anticancer effects and has used as a payload for antibody-drug conjugates. Our previous study have shown that although genetically engineered Actinosynnema pretiosum strains with enhanced UDPglucose (UDPG) biosynthesis displayed improved AP-3 production compared to the wild-type strain, the increase in yield was far from meeting the industrial demand. In this study, comparative metabolomics analysis complemented with quantitative real-time PCR analysis was performed for the wild-type strain and two mutants (OpgmOugp, Delta zwf Delta gnd) to identify possible metabolic bottlenecks and non-intuitive targets for further enhancement of AP-3 production. We observed that enhancing intracellular UDPG availability facilitated the accumulation of intracellular N-demethyl-AP-3 and AP-3, where the transporting of them outside the cell still needs to be developed. We also found that the UDPG biosynthesis was closely associated with the availability of fructose in the medium and a suitable fructose feeding strategy could promote the further improvement of AP-3 titer. In addition, pathway abundance analysis revealed that undesired fatty acid accumulation and down-regulation of amino acid metabolism may be unfavorable for ansamitocin biosynthesis in later stage of production. These results indicate that genetic modification of the UDPG biosynthetic pathways may have pleiotropic effects on AP-3 production. Efforts must be made to eliminate these newly identified metabolic bottlenecks to boost AP-3 production in A. pretiosum. (C) 2020, The Society for Biotechnology, Japan. All rights reserved.