Process of producing B-lactam antibiotics applying microorganisms with increased ligase activity
AssigneeGist-Brocades N.V.
InventorsSvend Kaasgaard, Klaus N. Kristiansen, Henrik M.o slashed.lgaard
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Abstract
The present invention relates to the biosynthesis of .beta.-lactam antibiotics. More specifically, the invention relates to processes of producing .beta.-lactam antibiotics in vivo and in vitro. Also contemplated is a novel enzyme capable of catalyzing certain steps involved in .beta.-lactam biosynthesis. Further, the invention relates to a DNA construct encoding the novel enzyme, a recombinant vector or transformation vehicle comprising the DNA construct, and finally a cell comprising the DNA construct or recombinant vector.
Claims32
An isolated acyl-coenzyme A synthetase obtained from Penicillium chrysogenum and which, in the synthesis of the corresponding acyl-coenzyme A esters, is at least 10 times more active towards phenoxyacetic acid than towards acetic acid, and is at least 10 times more active toward phenylacetic acid than towards acetic acid,
- wherein said synthetase comprises a property selected from the group consisting of (a) an apparent molecular weight of between 40,000 and 60,000 Daltons as determined by gel filtration, (b) an apparent activity optimum in the range of pH 7.0-9.0, and (c) an apparent activity optimum in the temperature range of 35-45.degree. C.
A method for the production of a .beta.-lactam antibiotic comprising incubating a carboxylic acid, coenzyme A, a synthetase exhibiting ligase activity according to clam 1, acyltransferase and a .beta.-lactam intermediate.
Dependencies are inferred from the claim text.