These authors contributed equally to this work.
Identification of OxyE as an Ancillary Oxygenase during Tetracycline Biosynthesis
Version of Record online: 26 MAY 2009
Copyright © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Volume 10, Issue 9, pages 1544–1550, June 15, 2009
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
- Issue online: 9 JUN 2009
- Version of Record online: 26 MAY 2009
- Manuscript Received: 7 MAR 2009
- NSF CBET. Grant Number: 0545860
- China Scholarship Council
- Nell I. Mondy Fellowship
- NMR spectroscopy;
- synthetic biology;
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.