Process for producing hydroxynitrile lyases
AssigneeJulich Chiral Solutions GmbH
InventorsFranz Effenberger, Harald Wajant, Siegfried Förster
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Abstract
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.
Claims20
A process for producing a hydroxynitrile lyase in dissolved form comprising:
- providing bacteria of the Escherichia coli species containing at least one gene, encoding a hydroxynitrile lyase of class II;
- precultivating the bacteria in a culture medium at 30 to 40 degrees Centigrade;
- inducing the bacteria with 1 to 100 μM IPTG (final concentration in the culture medium);
- cultivating the bacteria at 15 to 20 degrees Centigrade;
- lysing the bacteria;
- and
- isolating dissolved hydroxynitrile lyase.
Dependencies are inferred from the claim text.