Regulatory gene for clavulanic acid biosynthesis

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing oxygen-containing organic compound

Reexamination Certificate

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Details

C435S043000, C435S118000, C435S119000, C435S120000, C435S252300, C435S252350, C435S320100, C536S023700, C536S024100

Reexamination Certificate

active

06361982

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to recombinant DNA molecules and in particular to recombinant vectors for use in the transformation of a microbial host which contain inserted fragments carrying one or more genes involved in the regulation of clavulanic acid biosynthesis and cephamycin biosynthesis.
Progress in understanding the process of regulation of secondary metabolites produced by micro-organisms such as
Streptomyces clavuligerus
has been slow although it has been established that common control mechanisms may be responsible for the switching on of secondary metabolism in
S. coelicolor
. Two of the secondary metabolites produced in
S. clavuligerus
, clavulanic acid and cephamycin C, are important as antibiotics but appear to have totally unrelated pathways and nothing is currently known of how these pathways are regulated. In particular clavulanic acid is of great clinical value since it is a beta-lactamase inhibitor and protects beta-lactamase-labile beta-lactam antibiotics from degradation. Methods for increasing the yield (titre) of clavulanic acid in fermentation processes are therefore of considerable commercial importance.
2. Description of the Related Art
One approach to the problem of clavulanic acid yield improvement involves the use of recombinant DNA technology using
S. clavuligerus
as the host cell. Several enzymes are believed to be involved in clavulanic acid biosynthesis and the gene or genes encoding a clavaminic acid synthase, which converts one clavulanic acid intermediate, proclavaminic acid, into another one, clavaminic acid, have been cloned (European Patent Application, Publication Number 0 349 121). However, no genes concerned with the regulation of enzymes have so far been identified.


REFERENCES:
patent: 0 349 121 (1990-01-01), None
patent: 0 354 624 (1990-02-01), None
S. Horinouchi, et al., “Primary structure of AfsR, a global regulatory protein for secondary metabolite formation inStreptomyces coelicolorA3(2)”,Gene, 95 p.p. 49-56 (1990).
Juan F. Martin, “Clusters of genes for the biosynthesis of antibiotics: regulatory genes and overproduction of pharmaceuticals”,Chemical Abstracts, 117, Abstract No. 63796 (1992).

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