Maria successfully tackled a multi enzyme system which requires a NOX enzyme to recycle the NADH cofactor. Not a small battle when you require to maximise the air/oxygen content in a sealed system like a flow reactor. Very proud of this work!
Abstract: “As an alternative to classical synthetic approaches for the production of betazole drug, a one-pot biocatalytic system to this pharmaceutical molecule from its alcohol precursor has been developed. An ꙍ-transaminase, an alcohol dehydrogenase and a water-forming NADH oxidase for in-situ cofactor recycling have been combined to catalyse this reaction yielding 75% molar conversion in batch reaction with soluble enzymes. This multienzyme system was then co-immobilised through a newly established protocol for sequential functionalization of a methacrylate-based porous carrier to enable tailored immobilisation chemistries for each enzyme. This pluri-catalytic system has been set up in a continuous flow packed-bed reactor, yielding a space-time yield up to 2.59 g L-1 h-1 with 15 min residence with constant supply of oxygen for in-situ cofactor recycling through a segmented air-liquid flow. The addition of an in-line catch-and-release column afforded >80% product recovery.”
Read the full paper here