Secretory expression of cyclohexanone monooxygenase by methylotrophic yeast for efficient omeprazole sulfide bio-oxidation

Abstract Prochiral pyrmetazole can be asymmetrically oxidized into ( S )-omeprazole, a proton pump inhibitor that is used to treat gastroesophageal reflux, by an engineered cyclohexanone monooxygenase (CHMO Acineto - Mut) has high stereoselectivity. CHMO Mut produced heterologous expression in Escherichia coli , where it expressed intracellularly. Thus, isolating this useful biocatalyst requires tedious cell disruption and subsequent purification, which hinders its use for industrial purposes. Here, we report the extracellular production of methylotrophic yeast, Pichia pastoris, first time. The recombinant P. pastoris showed higher flavin occupation rate than E. was accompanied 3.2-fold increase catalytic efficiency. At density 150 g/L dry weight, achieved 1,700 U/L, representing approximately 85% total protein secreted fermentation broth. By directly employing pH adjusted supernatant as biocatalyst, were able almost completely transform 10 corresponding )-sulfoxide, with > 99% enantiomeric excess.