US 5,312,742 AGranted

Process for making L-phenyl acetyl carbinol (PAC), microorganisms for use in the process, and a method of preparing the microorganisms

AssigneeSynergen Associates, Inc.

InventorsRobert J. Seely, Donald L. Heefner, Robert V. Hageman, Michael J. Yarus, Sally A. Sullivan

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Abstract

Mutations are induced in a microorganism selected from the species Saccharomyces cerevisiae or from the species Candida flareri. The resulting mutants are cultured in the presence of a fermentation inhibitor, such as acetaldehyde, ephedrine or PAC-dione, to form colonies having resistance to the inhibitor. Cells from the colonies are isolated and tested for yield of phenyl acetyl carbinol (PAC) in a fermentation with benzaldehyde and pyruvate. Yeast cells from the colonies that produce elevated levels of PAC are selected for use in subsequent fermentations. PAC is useful as an intermediate in the preparation of l-ephedrine and d-pseudoephedrine, two well-known medicinal chemicals.

Claims7

Dependent claims

A process for the production of L-phenyl acetyl carbinol (PAC), which comprises,

providing an aqueous fermentation medium containing a submerged culture of a microorganism and a source of assimilable carbon;
cultivating said organism under anaerobic or oxygen limited conditions in the presence of benzaldehyde and pyruvate to form PAC in a concentration of at least about 12 g/L in said fermentation medium;
and
wherein said organism is a mutant of a parent strain of Saccharomyces cerevisiae or a mutant of a parent strain of Candida flareri, and said mutant is more resistant to aldehyde inhibition and produces less acetaldehyde and a higher concentration of PAC in the fermentation medium than a corresponding fermentation using the parent strain under otherwise identical conditions.

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Dependencies are inferred from the claim text.