Degussa Aktiengesellschaft

Dsm Fine Chemicals Austria Gmbh

Julich Chiral Solutions Gmbh

Harald Wajant

Franz Effenberger

Helmut Schwab

Christoph Kratky

Ernst Steiner

Siegfried Förster

Holger Buhler

Siegfried F&ouml;rster

Peter Lauble

Ulrike Wagner

Jurgen Roos

The present invention relates to an improved process for producing hydroxynitrile lyases. A process is provided whereby hydroxynitrile lyases may be produced by cultivation in bacteria or suitable host cells. Cells containing a gene encoding for a hydroxynitrile lyase may be precultivated, induced with a low concentration of IPTG, cultured, and lysed. The process allows for the production of relatively large quantities of hydroxynitrile lyases in a dissolved, native, and active form, and without the presence of substantial amounts of inclusion bodies, and thus allowing for the production of the lyases without requiring expensive and time consuming renaturation steps.

Process for producing hydroxynitrile lyases

(S)-Hydroxynitrile lyases which have an improved substrate acceptance and which are derived from the,and,(S)-hydroxynitrile lyases, wherein one or more bulky amino acid residues within the hydrophobic channel leading to the active center have been replaced with less bulky amino acid residues.

(S)-hydroxynitrile lyases with improved substrate acceptance and their use

A method for producing (S)-cyanohydrins of general formula ##STR1## in which R and R' independently represent (1) hydrogen; (2) a substituted or unsubstituted saturated alkyl group which may include amine, imine, hydroxy, C.sub.1 -C.sub.8 -alkoxy, halogen, carboxyl, C.sub.3 -C.sub.20 -cycloalkyl groups and/or a N,O,S-heteroatom-substituted aromatic ring as substituent; (3) a substituted or unsubstituted, singly or multiply unsaturated alkenyl- or alkinyl group which may include one or several amine, imine, hydroxy, C.sub.1 -C.sub.8 -alkoxy, halogen, carboxyl, C.sub.3 -C.sub.20 -cycloalkyl groups and/or one optionally N,O,S-heteroatom-substituted aromatic ring as substituents; (4) a substituted or unsubstituted aromatic or heteroaromatic group. The method includes an enzyme-catalyzed conversion of carbonyl compounds of general formula II ##STR2## with hydrogen cyanide or a substance supplying hydrogen cyanide or CN- for the conversion in the presence of an amount of an immobilized (S)-oxynitrilase which catalyzes the conversion. The use of nitrocellulose as carrier for the immobilized (S)-oxynitrilase results in a distinct increase of the yield of (S)-cyanohydrin and of the enantiomeric excess compared to the traditional cellulose carrier material.

Method of producing (S)-cyanohydrins

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Leinfelden, Germany

Harald Wajant