ENGINEERING PICHIA-PASTORIS FOR BIOCATALYSIS - COPRODUCTION OF 2 ACTIVE ENZYMES

High levels of active glycolate oxidase from spinach (GO) and active c atalase T from Saccharomyces cerevisiae (catT) have been co-produced i n the methylotrophic yeast Pichia pastoris (Pp). In sequential rounds of transformation using two selectable markers, multiple copies of the genes encoding GO and catT were integrated into the Pp chromosome und er control of the methanol inducible alcohol oxidase I promoter, resul ting in a strain designated MSP8.6. MSP8.6 is a second-generation bioc atalyst used for the conversion of glycolate to glyoxylate in the pres ence of a reaction component which inhibits endogenous Pp catalase. Th is work demonstrates a significant advance in the utility of recombina nt Pp for commercial bioprocess development. (C) 1997 Elsevier Science B.V.