1887

Abstract

The genetic organization of the region upstream of the gene of the clavulanic acid biosynthetic gene cluster of has been determined. Sequence analysis of a 121 kb region revealed the presence of 10 ORFs whose putative functions, according to database searches, are discussed. Three co-transcriptional units are proposed: ORF10–11, ORF12–13 and ORF15–16–17–18. Potential transcriptional terminators were identified downstream of ORF11 () and ORF15. Targeted disruption of ORF10 () gave rise to transformants unable to produce clavulanic acid, but with a considerably higher production of cephamycin C. Transformants inactivated at ORF14 had a remarkably lower production of clavulanic acid and similar production of cephamycin C. Significant improvements of clavulanic acid production, associated with a drop in cephamycin C biosynthesis, were obtained with transformants of harbouring multiple copies of plasmids carrying different constructions from the ORF10–14 region. This information can be used to guide strain improvement programs, blending random mutagenesis and molecular cloning, to optimize the yield of clavulanic acid.

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2002-05-01
2020-09-18
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