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.
MeSH Terms
Anti-Bacterial Agents/biosynthesis
Blotting, Southern
Chromosome Walking
Cloning, Molecular
Genes, Bacterial
L-Lysine 6-Transaminase
Multigene Family
Restriction Mapping
Streptomyces/enzymology,genetics
Transaminases/genetics,metabolism
beta-Lactams
Chemicals
Anti-Bacterial Agents
beta-Lactams
Transaminases
L-Lysine 6-Transaminase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Madduri K
Department of Microbiology, Dalhousie University, Halifax, Nova Scotia, Canada.
Stuttard C
Vining L C
References (12)
12 references, click to expand
-
Lysine catabolism in Streptomyces spp. is primarily through cadaverine: beta-lactam producers also make alpha-aminoadipate.
J Bacteriol. 1989 Jan;171(1):299-302
PMID: 2492500
-
Cloning, characterization, and expression in Escherichia coli of the Streptomyces clavuligerus gene encoding deacetoxycephalosporin C synthetase.
J Bacteriol. 1989 Feb;171(2):754-60
PMID: 2644235
-
Cloning and characterization of beta-lactam biosynthetic genes.
Mol Microbiol. 1989 May;3(5):689-95
PMID: 2668696
-
Beta-lactam antibiotic biosynthetic genes have been conserved in clusters in prokaryotes and eukaryotes.
EMBO J. 1990 Mar;9(3):741-7
PMID: 2107074
-
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):3952-8
PMID: 1694525
-
Chimeric single-stranded DNA phage-plasmid cloning vectors.
Biotechnology. 1988;10:85-102
PMID: 3061525
-
Plasmids, recombination and chromosome mapping in Streptomyces lividans 66.
J Gen Microbiol. 1983 Jul;129(7):2257-69
PMID: 6631413
-
Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans.
J Gen Microbiol. 1983 Sep;129(9):2703-14
PMID: 6313861
-
An improved technique for the efficient construction of gene libraries by partial filling-in of cohesive ends.
Gene. 1986;42(1):119-23
PMID: 3013727
-
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
PMID: 3038334
-
Cloning and nucleotide sequence determination of the isopenicillin N synthetase gene from Streptomyces clavuligerus.
Gene. 1988;62(2):187-96
PMID: 3130293
-
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):1251-8
PMID: 1700062