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Identification of OxyE as an Ancillary Oxygenase during Tetracycline Biosynthesis

  1. Peng Wang1,2,†,
  2. Wenjun Zhang2,†,
  3. Jixun Zhan Dr.2,3,
  4. Yi Tang Prof.2

Article first published online: 26 MAY 2009

DOI: 10.1002/cbic.200900122

Issue

ChemBioChem

ChemBioChem

Volume 10, Issue 9, pages 1544–1550, June 15, 2009

Additional Information

How to Cite

Wang, P., Zhang, W., Zhan, J. and Tang, Y. (2009), Identification of OxyE as an Ancillary Oxygenase during Tetracycline Biosynthesis. ChemBioChem, 10: 1544–1550. doi: 10.1002/cbic.200900122

Author Information

  1. 1

    Laboratory of Microbial Metabolism and School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200030 (China)

  2. 2

    Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, 420 Westwood Plaza, Los Angeles, CA 90095 (USA), Fax: (+1) 310-206-4107

  3. 3

    Present address: Department of Biological and Irrigation Engineering, Utah State University, 105 Old Main Hill Logan, UT 84322-4105(USA)

  1. †

    These authors contributed equally to this work.

  1. †

    These authors contributed equally to this work.

Publication History

  1. Issue published online: 9 JUN 2009
  2. Article first published online: 26 MAY 2009
  3. Manuscript Received: 7 MAR 2009

Funded by

  • NSF CBET. Grant Number: 0545860
  • China Scholarship Council
  • Nell I. Mondy Fellowship

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Keywords:

  • metabolism;
  • NMR spectroscopy;
  • oxygenase;
  • polyketides;
  • synthetic biology;
  • tetracycline

Abstract

Thumbnail image of graphical abstract

Ancillary oxygenase: OxyE is identified as a likely ancillary C-4 hydroxylase used during oxytetracycline biosynthesis in Streptomyces rimosus. The synergistic actions of oxygenases OxyE and OxyL ensure complete oxidative tailoring and prevent irreversible shunt modifications of the biosynthetic intermediate.

The double hydroxylation of 6-pretetramid to 4-keto-anhydrotetracycline is a key tailoring reaction during the biosynthesis of the broad-spectrum antibiotic tetracyclines. It has been shown previously by heterologous reconstitution that OxyL is a dioxygenase and is the only enzyme required to catalyze the insertion of oxygen atoms at the C-12a and C-4 positions. We report here that OxyE, a flavin adenine dinucleotide (FAD)-dependent hydroxylase homologue, is an ancillary mono-oxygenase for OxyL during oxytetracycline biosynthesis in Streptomyces rimosus. By using both gene disruption and heterologous reconstitution approaches, we demonstrated that OxyE plays a nonessential, but important role in oxytetracycline biosynthesis by serving as a more efficient C-4 hydroxylase. In addition, we demonstrated that partially oxidized biosynthetic intermediates can undergo various glycosylation modifications in S. rimosus. Our results indicate that the synergistic actions of OxyE and OxyL in the double hydroxylation step prevent accumulation of shunt products during oxytetracycline biosynthesis in S. rimosus.

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