Enantioselective oxidation of 2-hydroxy carboxylic acids by glycolate oxidase and catalase coexpressed in methylotrophic Pichia pastoris.

摘要: Glycolate oxidase (GO; (S)-2-hydroxyacid oxidase, EC 1.1.3.15) is a flavin mononucleotide (FMN)-dependent enzyme, which catalyzes the oxidation of 2-hydroxy carboxylic acids to corresponding 2-keto acids. Catalase has been used as cocatalyst decompose hydrogen peroxide produced in reaction, thus limiting peroxide-based side reactions and GO deactivation. from spinach catalase T Saccharomyces cerevisiae previously coexpressed Pichia pastoris strain NRRL Y-21001, was permeabilized for 3-phenyllactic acid, 3-indolelactic 3-chlorolactic 2-hydroxybutanoic 2-hydroxydecanoic acid demonstrate high degree selectivity (S)-enantiomers, leaving (R)-isomers intact. The rates ranged 1.3 120.0%, relative lactic pyruvic acid. best substrates were (110%) (120%). Oxidation carried out with (R)-, (S)-, (RS)-3-phenyllactic (RS)-lactic (RS)-2-hydroxybutanoic 500 mL scale characterize products stoichiometry reaction. All (RS)- (S)-2-hydroxy at close theoretical yield 1-9 h. (R)-3-Phenyllactic not oxidized over period 9 Addition exogenous FMN required this using double recombinant whole cells. As absolutely specific it can be resolution racemic (R)-2-hydroxy well production This first report on broad range by GO.