Applied Microbiology and Biotechnology

, Volume 100, Issue 5, pp 2257–2266

In vivo investigation to the macrolide-glycosylating enzyme pair DesVII/DesVIII in Saccharopolyspora erythraea

  • Hang Wu
  • Weiwei Li
  • Chen Xin
  • Congming Zhang
  • Yansheng Wang
  • Shaohua Ren
  • Min Ren
  • Wei Zhao
  • Li Yuan
  • Zhongdong Xu
  • Hualing Yuan
  • Ming Geng
  • Lixin Zhang
  • David T. Weaver
  • Buchang Zhang
Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-015-7036-9

Cite this article as:
Wu, H., Li, W., Xin, C. et al. Appl Microbiol Biotechnol (2016) 100: 2257. doi:10.1007/s00253-015-7036-9

Abstract

Glycosyltransferase DesVII and its auxiliary partner DesVIII from Streptomyces venezulae, homologs of EryCIII and EryCII in Saccharopolyspora erythraea, have previously been demonstrated to be flexible on their substrates in vitro. Herein, we investigated their in vivo function by interspecies complementation in the mutant strains of Sac. erythraea A226. As desVII and desVIII were concomitantly expressed in the ΔeryCIII mutant, the erythromycin A (Er-A) production was restored. Interestingly, co-expression of desVII and desVIII in the ΔeryBV mutant exhibited an increased Er-A yield by 15 % in comparison to A226. Hence, DesVII/DesVIII not only replaced EryCIII to upload D-desosamine to C5 position of 3-O-mycarosyl erythronolide B (MEB) but also in vivo attached L-mycarose, not D-desosamine to C3 position of erythronolide B (EB) with a higher activity than EryBV. Furthermore, expression of desVII in ΔeryCIII and ΔeryBV-CIII partially restored the Er-A production; however, no Er-A was detected while desVII was expressed in ΔeryBV. It was implicated that DesVII coupled with EryCII to form the DesVII/EryCII complex for attaching above two deoxysugars in the absence of EryCIII in Sac. erythraea. In addition, when desVII and desVIII were co-expressed in ΔeryBV-CII, Er-A was recovered with a lower yield than ΔeryBV-CIII. Our study presents an opportunity with Sac. erythraea as a cell factory for macrolide glycodiversification.

Keywords

Saccharopolyspora erythraea Glycosyltransferase DesVII/DesVIII EryCII/EryCIII EryBV 

Supplementary material

253_2015_7036_MOESM1_ESM.pdf (269 kb)
ESM 1(PDF 268 kb)

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Hang Wu
    • 1
  • Weiwei Li
    • 1
  • Chen Xin
    • 1
  • Congming Zhang
    • 1
  • Yansheng Wang
    • 1
  • Shaohua Ren
    • 1
  • Min Ren
    • 1
  • Wei Zhao
    • 1
  • Li Yuan
    • 1
  • Zhongdong Xu
    • 3
  • Hualing Yuan
    • 3
  • Ming Geng
    • 3
  • Lixin Zhang
    • 1
    • 2
  • David T. Weaver
    • 1
  • Buchang Zhang
    • 1
  1. 1.Institute of Health Sciences, School of Life SciencesAnhui UniversityHefeiChina
  2. 2.CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of MicrobiologyChinese Academy of SciencesBeijingChina
  3. 3.School of Life SciencesHefei Normal UniversityHefeiChina

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