Chinese Journal of Organic Chemistry ›› 2020, Vol. 40 ›› Issue (11): 3828-3836.DOI: 10.6023/cjoc202006059 Previous Articles     Next Articles

Special Issue: 创刊四十周年专辑


范亚飞a, 张鄂a, 郭恒b, 牟柠a, 陈单丹b, 王文贵a, 王守锋a, 刘文b,c   

  1. a 济南大学化学化工学院 山东省氟化学化工材料重点实验室 济南 250022;
    b 中国科学院上海有机化学研究所 生命有机化学国家重点实验室 上海 200032;
    c 中国科学院上海有机化学研究所 湖州生物制造中心 浙江湖州 313000
  • 收稿日期:2020-06-28 修回日期:2020-08-06 发布日期:2020-09-09
  • 通讯作者: 王守锋, 刘文;
  • 基金资助:

Insights into the Substrate Tolerance of Enzymes Involved in the Nosiheptide Biosynthesis Pathway Based on Indolic Acid Moiety

Fan Yafeia, Zhang Ea, Guo Hengb, Mu Ninga, Chen Dandanb, Wang Wenguia, Wang Shoufenga, Liu Wenb,c   

  1. a Shandong Provincial Key Laboratory of Fluorine Chemistry and Chemical Materials, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022;
    b State Key Laboratory of Bioorganic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032;
    c Huzhou Center of Bio-Synthetic Innovation, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Huzhou, Zhejiang 313000
  • Received:2020-06-28 Revised:2020-08-06 Published:2020-09-09
  • Supported by:
    Project supported by the National Natural Science Foundation of China (Nos. 31972850, 21750004, 21520102004), the Shandong Key Research Program (No. 2019GSF108223), the Chinese Academy of Sciences (Nos. QYZDJ-SSW-SLH037, XDB20020200), the Science and Technology Commission of Shanghai Municipality (No. 17JC1405100), the Youth Innovation Promotion Association of the Chinese Academy of Sciences (No. 2017303), the State Key Laboratory of Microbial Technology Open Projects Fund (No. M2020-05) and the K. C. Wong Education Foundation.

As a typical representative of thiopeptide antibiotics, nosiheptide (NOS) possesses very good antibacterial activity. However, due to poor water solubility and low bioavailability, its clinical application is hampered. Due to its complex structure, it is difficult to obtain analogues with improved physical and chemical properties via total chemical synthesis. Based on the previous studies on the biosynthesis of nosiheptide, the side-ring 3-methyl-2-indoleic acid (MIA) analogues were used as chemical small molecule probes to explore the substrate tolerance of enzymes involved in NOS biosynthesis pathway in NOS-producing bacteria via the co-fermentation of probe molecules with mutant strain and the combination of high resolution mass spectrometry data of fermentation products. The results showed that enzymes involved in NOS biosynthesis pathway had a considerable tolerance to MIA analogues substituted by F, Cl and CH3, however, MIA analogues substituted by large steric hindrance group, such as NO2, CF3 and Ph, were not tolerated. The position, the size and the property of the substituted groups of MIA also affected the steps of identification, transport and upload of the related enzymes involved in NOS biosynthesis. The present study not only explored the substrate tolerance of enzymes involved in NOS biosynthesis pathway, but also was expected to obtain NOS analogues via biosynthetic pathway engineering. What’s more, it provides valuable information for using directed evolution technology to improve the substrate tolerance of enzymes in the rate-limiting steps of NOS biosynthesis and to expand the use of NOS-producing bacteria to obtain more analogues.

Key words: nosiheptide, 3-methyl-2-indolic acid, mutational biosynthesis, chemical small molecule probe, substrate tolerance of enzymes