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. 1991 Feb;173(3):985–988. doi: 10.1128/jb.173.3.985-988.1991

Cloning and location of a gene governing lysine epsilon-aminotransferase, an enzyme initiating beta-lactam biosynthesis in Streptomyces spp.

K Madduri 1, C Stuttard 1, L C Vining 1
PMCID: PMC207215  PMID: 1991735

Abstract

In actinomycetes that produce beta-lactam antibiotics of the cephem type, lysine epsilon-aminotransferase is the initial enzyme in the conversion of lysine to alpha-aminoadipic acid. We used a two-stage process ("chromosome walking") to screen a lambda library of Streptomyces clavuligerus genomic DNA for fragments that expressed lysine epsilon-aminotransferase activity in S. lividans. Restriction analysis of the cloned DNA confirmed the location of the putative lat gene within the cluster of beta-lactam biosynthesis genes, roughly midway between pcbC, the structural gene for isopenicillin N synthetase, and the putative cefE gene encoding deacetoxycephalosporin C synthetase.

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Selected References

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  1. Hopwood D. A., Kieser T., Wright H. M., Bibb M. J. Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol. 1983 Jul;129(7):2257–2269. doi: 10.1099/00221287-129-7-2257. [DOI] [PubMed] [Google Scholar]
  2. Katz E., Thompson C. J., Hopwood D. A. Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol. 1983 Sep;129(9):2703–2714. doi: 10.1099/00221287-129-9-2703. [DOI] [PubMed] [Google Scholar]
  3. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. 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]
  7. Leskiw B. K., Mevarech M., Barritt L. S., Jensen S. E., Henderson D. J., Hopwood D. A., Bruton C. J., Chater K. F. Discovery of an insertion sequence, IS116, from Streptomyces clavuligerus and its relatedness to other transposable elements from actinomycetes. J Gen Microbiol. 1990 Jul;136(7):1251–1258. doi: 10.1099/00221287-136-7-1251. [DOI] [PubMed] [Google Scholar]
  8. Madduri K., Stuttard C., Vining L. C. Lysine catabolism in Streptomyces spp. is primarily through cadaverine: beta-lactam producers also make alpha-aminoadipate. J Bacteriol. 1989 Jan;171(1):299–302. doi: 10.1128/jb.171.1.299-302.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mead D. A., Kemper B. Chimeric single-stranded DNA phage-plasmid cloning vectors. Biotechnology. 1988;10:85–102. doi: 10.1016/b978-0-409-90042-2.50010-6. [DOI] [PubMed] [Google Scholar]
  10. Miller J. R., Ingolia T. D. Cloning and characterization of beta-lactam biosynthetic genes. Mol Microbiol. 1989 May;3(5):689–695. doi: 10.1111/j.1365-2958.1989.tb00217.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Zabarovsky E. R., Allikmets R. L. An improved technique for the efficient construction of gene libraries by partial filling-in of cohesive ends. Gene. 1986;42(1):119–123. doi: 10.1016/0378-1119(86)90158-7. [DOI] [PubMed] [Google Scholar]

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