Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1991 Jan;173(1):398–400. doi: 10.1128/jb.173.1.398-400.1991

Cloning and sequencing of the beta-lactam hydroxylase gene (cefF) from Streptomyces clavuligerus: gene duplication may have led to separate hydroxylase and expandase activities in the actinomycetes.

S Kovacevic 1, J R Miller 1
PMCID: PMC207200  PMID: 1987130

Abstract

The deacetylcephalosporin C synthetase (hydroxylase) gene from Streptomyces clavuligerus has been cloned and sequenced. The open reading frame codes for a protein with an Mr of 34,584. The hydroxylase gene (cefF) is closely linked to the epimerase gene (cefD) and the expandase gene (cefE) and is transcribed in the opposite orientation. The hydroxylase and expandase genes are 59 and 71% identical at the amino acid and DNA levels, respectively. cefE and cefF may have arisen from a gene duplication in the actinomycetes.

Full text

PDF
398

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cantwell C. A., Beckmann R. J., Dotzlaf J. E., Fisher D. L., Skatrud P. L., Yeh W. K., Queener S. W. Cloning and expression of a hybrid Streptomyces clavuligerus cefE gene in Penicillium chrysogenum. Curr Genet. 1990 Mar;17(3):213–221. doi: 10.1007/BF00312612. [DOI] [PubMed] [Google Scholar]
  2. Dotzlaf J. E., Yeh W. K. Copurification and characterization of deacetoxycephalosporin C synthetase/hydroxylase from Cephalosporium acremonium. J Bacteriol. 1987 Apr;169(4):1611–1618. doi: 10.1128/jb.169.4.1611-1618.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ingolia T. D., Queener S. W. Beta-lactam biosynthetic genes. Med Res Rev. 1989 Apr-Jun;9(2):245–264. doi: 10.1002/med.2610090206. [DOI] [PubMed] [Google Scholar]
  4. Jensen S. E., Westlake D. W., Wolfe S. Deacetoxycephalosporin C synthetase and deacetoxycephalosporin C hydroxylase are two separate enzymes in Streptomyces clavuligerus. J Antibiot (Tokyo) 1985 Feb;38(2):263–265. doi: 10.7164/antibiotics.38.263. [DOI] [PubMed] [Google Scholar]
  5. Kovacevic S., Tobin M. B., Miller J. R. The beta-lactam biosynthesis genes for isopenicillin N epimerase and deacetoxycephalosporin C synthetase are expressed from a single transcript in Streptomyces clavuligerus. J Bacteriol. 1990 Jul;172(7):3952–3958. doi: 10.1128/jb.172.7.3952-3958.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kovacevic S., Weigel B. J., Tobin M. B., Ingolia T. D., Miller J. R. Cloning, characterization, and expression in Escherichia coli of the Streptomyces clavuligerus gene encoding deacetoxycephalosporin C synthetase. J Bacteriol. 1989 Feb;171(2):754–760. doi: 10.1128/jb.171.2.754-760.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Leskiw B. K., Aharonowitz Y., Mevarech M., Wolfe S., Vining L. C., Westlake D. W., Jensen S. E. Cloning and nucleotide sequence determination of the isopenicillin N synthetase gene from Streptomyces clavuligerus. Gene. 1988;62(2):187–196. doi: 10.1016/0378-1119(88)90557-4. [DOI] [PubMed] [Google Scholar]
  8. Piret J., Resendiz B., Mahro B., Zhang J. Y., Serpe E., Romero J., Connors N., Demain A. L. Characterization and complementation of a cephalosporin-deficient mutant of Streptomyces clavuligerus NRRL 3585. Appl Microbiol Biotechnol. 1990 Feb;32(5):560–567. doi: 10.1007/BF00173728. [DOI] [PubMed] [Google Scholar]
  9. Samson S. M., Chapman J. L., Belagaje R., Queener S. W., Ingolia T. D. Analysis of the role of cysteine residues in isopenicillin N synthetase activity by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5705–5709. doi: 10.1073/pnas.84.16.5705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Scheidegger A., Küenzi M. T., Nüesch J. Partial purification and catalytic properties of a bifunctional enzyme in the biosynthetic pathway of beta-lactams in Cephalosporium acremonium. J Antibiot (Tokyo) 1984 May;37(5):522–531. doi: 10.7164/antibiotics.37.522. [DOI] [PubMed] [Google Scholar]
  11. Singh P. D., Ward P. C., Wells J. S., Ricca C. M., Trejo W. H., Principe P. A., Sykes R. B. Bacterial production of deacetoxycephalosporin C. J Antibiot (Tokyo) 1982 Oct;35(10):1397–1399. doi: 10.7164/antibiotics.35.1397. [DOI] [PubMed] [Google Scholar]
  12. Smith D. J., Burnham M. K., Bull J. H., Hodgson J. E., Ward J. M., Browne P., Brown J., Barton B., Earl A. J., Turner G. Beta-lactam antibiotic biosynthetic genes have been conserved in clusters in prokaryotes and eukaryotes. EMBO J. 1990 Mar;9(3):741–747. doi: 10.1002/j.1460-2075.1990.tb08168.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES