Process for the preparation of defective mutants of microorganisms
AssigneeHenkel Kommanditgesellschaft auf Aktien
InventorsFrank F. Hill, Joachim Schindler, Rolf Schmid, Wolfgang Preuss, Alfred Struve
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Abstract
For the production of 17-C-steroid-.alpha.-propionic acid compounds by microbial side chain degradation on 17-C-side chain steroid substrates wild strain microorganisms selected in predetermined manner are subjected to a multi-stage mutation and selection. The resultant mutant strains are capable of supplying the desired compounds, e.g. .DELTA.-4 BNC and/or .DELTA.-1,4 BNC in high yields even in the absence of inhibitors arresting the growth and/or degradation of the steroid ring. Wild strains and mutants particularly suitable, and their choice, production and use, are described.
Claims12
A process for the production of biologically pure microorganism defective mutants which are capable of producing 17-C-steroid-alpha-propionic acid compounds including 3-oxo-pregna-4-ene-20 carboxylic acid and 3-oxo-1,4-diene-20-carboxylic acid and mixtures thereof from 17-C-side chain steroid substrates by selective side chain degradation under aerobic conditions without substantial steroid ring degradation in the absence of ring degradation inhibitors, where the biologically pure microorganism defective mutants are obtained from a population of mutants containing undesired accompanying mutant strains wherein
- (a) a microorganism wild strain which is capable of growing on sterol compounds as a sole source of carbon is isolated and cultivated,
- (b) the wild strain is treated by a known mutation treatment, to give the mutant population,
- (c) the mutant population obtained from (b) is cultivated on separation medium on which the mutants capable of degrading the sterol compounds only to the C-17-steroid-alpha-propionic acid do not substantially grow, and
- (d) the mutant micrioorganisms capable of growing in (c) are killed off due to their growth or through the use of antibiotics or radioactive media thereby giving the biologically pure defective mutant strain and,
- (e) the biologically pure mutant strains that do not substantially grow are isolated and preserved.
A biologically pure culture of defective mutant strains selected from the group consisting of ATCC 31456, ATCC 31457, ATCC 31459 and ATCC 31460.
Dependencies are inferred from the claim text.