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Applied Microbiology and Biotechnology

, Volume 101, Issue 13, pp 5291–5300 | Cite as

Engineered jadomycin analogues with altered sugar moieties revealing JadS as a substrate flexible O-glycosyltransferase

  • Liyuan Li
  • Guohui Pan
  • Xifen Zhu
  • Keqiang Fan
  • Wubin Gao
  • Guomin Ai
  • Jinwei Ren
  • Mingxin Shi
  • Carlos Olano
  • José A. Salas
  • Keqian Yang
Biotechnologically relevant enzymes and proteins

Abstract

Glycosyltransferases (GTs)-mediated glycodiversification studies have drawn significant attention recently, with the goal of generating bioactive compounds with improved pharmacological properties by diversifying the appended sugars. The key to achieving glycodiversification is to identify natural and/or engineered flexible GTs capable of acting upon a broad range of substrates. Here, we report the use of a combinatorial biosynthetic approach to probe the substrate flexibility of JadS, the GT in jadomycin biosynthesis, towards different non-native NDP-sugar substrates, enabling us to identify six jadomycin B analogues with different sugar moieties. Further structural engineering by precursor-directed biosynthesis allowed us to obtain 11 new jadomycin analogues. Our results for the first time show that JadS is a flexible O-GT that can utilize both l- and d- sugars as donor substrates, and tolerate structural changes at the C2, C4 and C6 positions of the sugar moiety. JadS may be further exploited to generate novel glycosylated jadomycin molecules in future glycodiversification studies.

Keywords

Jadomycin JadS Glycosyltransferase Glycosylation Precursor-directed biosynthesis Cytotoxicity 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant: 31130001). We thank Ajeeth Adhikari at The Scripps Research Institute for critical reading of this manuscript, and Dr. Chin-Yuan Chang at The Scripps Research Institute for helping analyze the structure model of JadS.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2017_8256_MOESM1_ESM.pdf (1.9 mb)
ESM 1 (PDF 1977 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.State Key Laboratory of Microbial ResourcesInstitute of Microbiology, Chinese Academy of SciencesBeijingPeople’s Republic of China
  2. 2.State Key Laboratory of MycologyInstitute of Microbiology, Chinese Academy of SciencesBeijingPeople’s Republic of China
  3. 3.Departamento de Biología Funcional e Instituto Universitario de Oncología del Principado de Asturias (I.U.O.P.A)Universidad de OviedoOviedoSpain

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