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PMID: 10869089 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Expansion of the clavulanic acid gene cluster: identification and in vivo functional analysis of three new genes required for biosynthesis of clavulanic acid by Streptomyces clavuligerus.

Journal of bacteriology ·Vol. 182 ·No. 14 ·2000-07-00 ·Pages 4087-95

Li R, Khaleeli N, Townsend CA

Abstract

Clavulanic acid is a potent inhibitor of beta-lactamase enzymes and is of demonstrated value in the treatment of infections by beta-lactam-resistant bacteria. Previously, it was thought that eight contiguous genes within the genome of the producing strain Streptomyces clavuligerus were sufficient for clavulanic acid biosynthesis, because they allowed production of the antibiotic in a heterologous host (K. A. Aidoo, A. S. Paradkar, D. C. Alexander, and S. E. Jensen, p. 219-236, In V. P. Gullo et al., ed., Development in industrial microbiology series, 1993). In contrast, we report the identification of three new genes, orf10 (cyp), orf11 (fd), and orf12, that are required for clavulanic acid biosynthesis as indicated by gene replacement and trans-complementation analysis in S. clavuligerus. These genes are contained within a 3.4-kb DNA fragment located directly downstream of orf9 (cad) in the clavulanic acid cluster. While the orf10 (cyp) and orf11 (fd) proteins show homologies to other known CYP-150 cytochrome P-450 and [3Fe-4S] ferredoxin enzymes and may be responsible for an oxidative reaction late in the pathway, the protein encoded by orf12 shows no significant similarity to any known protein. The results of this study extend the biosynthetic gene cluster for clavulanic acid and attest to the importance of analyzing biosynthetic genes in the context of their natural host. Potential functional roles for these proteins are proposed.

MeSH Terms
Amino Acid Sequence Base Sequence Clavulanic Acid/biosynthesis Cytochrome P-450 Enzyme System/genetics Ferredoxins/genetics Genes, Bacterial Genetic Complementation Test Molecular Sequence Data Multigene Family Mutagenesis, Insertional Open Reading Frames Restriction Mapping Sequence Analysis, DNA Streptomyces/genetics beta-Lactam Resistance beta-Lactamase Inhibitors
Chemicals
Ferredoxins beta-Lactamase Inhibitors Clavulanic Acid Cytochrome P-450 Enzyme System
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Li R
Department of Chemistry, The Johns Hopkins University, Baltimore, MD 21218, USA.
Khaleeli N
Townsend C A
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.182.14.4087-4095.2000
Published
2000-07-00
Pages
4087-95
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC94596
Subset
IM
Grants
NIAID NIH HHS · R01 AI014937 · United States
NIAID NIH HHS · R37 AI014937 · United States
NIAID NIH HHS · R56 AI014937 · United States
NIAID NIH HHS · AI 14937 · United States
Databases
GENBANK
AF200819
External Links
PubMed source DOI: 10.1128/JB.182.14.4087-4095.2000 PMC Full Text View journal details
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