This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Binnie, C Articles by Butler, M J Search for Related Content PubMed PubMed Citation Articles by Binnie, C Articles by Butler, M J

research-article

Cloning and heterologous expression in Streptomyces lividans of Streptomyces rimosus genes involved in oxytetracycline biosynthesis.

International Process Development Group, Pfizer Limited, Sandwich, Kent, United Kingdom.

ABSTRACT

The anhydrotetracycline (ATC) oxygenase enzyme which carries out the conversion of ATC to dehydrotetracycline was purified and the N-terminal amino acid sequence was determined. The sequence displays a significant similarity to that of the p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens. This is consistent with the activity of the oxygenase, i.e., addition of a hydroxyl moiety to an aromatic ring structure. Oligonucleotide probes were designed and used to clone the corresponding fragment of chromosomal DNA from Streptomyces rimosus. This DNA fragment was used to screen a cosmid library, allowing the isolation of flanking DNA sequences. Surprisingly, the gene was located within the previously cloned cluster of genes involved in the synthesis of the biosynthetic intermediate ATC and not as had been expected (P. M. Rhodes, N. Winskill, E. J. Friend, and M. Warren, J. Gen. Microbiol. 124:329-338, 1981) at a separate locus on the other side of the chromosome. Subcloning of an appropriate DNA fragment from one of the cosmid clones onto pIJ916 produced Streptomyces lividans transformants which synthesized oxytetracycline.

This article has been cited by other articles:

• Petkovic, H., Cullum, J., Hranueli, D., Hunter, I. S., Peric-Concha, N., Pigac, J., Thamchaipenet, A., Vujaklija, D., Long, P. F. (2006). Genetics of Streptomyces rimosus, the Oxytetracycline Producer. Microbiol. Mol. Biol. Rev. 70: 704-728 [Abstract] [Full Text]  
• Zhang, W., Ames, B. D., Tsai, S.-C., Tang, Y. (2006). Engineered Biosynthesis of a Novel Amidated Polyketide, Using the Malonamyl-Specific Initiation Module from the Oxytetracycline Polyketide Synthase. Appl. Environ. Microbiol. 72: 2573-2580 [Abstract] [Full Text]  
• Peric-Concha, N., Borovicka, B., Long, P. F., Hranueli, D., Waterman, P. G., Hunter, I. S. (2005). Ablation of the otcC Gene Encoding a Post-polyketide Hydroxylase from the Oxytetracyline Biosynthetic Pathway in Streptomyces rimosus Results in Novel Polyketides with Altered Chain Length. J. Biol. Chem. 280: 37455-37460 [Abstract] [Full Text]  
• Petkovic, H., Thamchaipenet, A., Zhou, L.-H., Hranueli, D., Raspor, P., Waterman, P. G., Hunter, I. S. (1999). Disruption of an Aromatase/Cyclase from the Oxytetracycline Gene Cluster of Streptomyces rimosus Results in Production of Novel Polyketides with Shorter Chain Lengths. J. Biol. Chem. 274: 32829-32834 [Abstract] [Full Text]  
• McDowall, K. J., Thamchaipenet, A., Hunter, I. S. (1999). Phosphate Control of Oxytetracycline Production by Streptomyces rimosus Is at the Level of Transcription from Promoters Overlapped by Tandem Repeats Similar to Those of the DNA-Binding Sites of the OmpR Family. J. Bacteriol. 181: 3025-3032 [Abstract] [Full Text]