The pivaloyloxymethyl and butanoyloxymethyl derivatives of tert-butanol and linalool (1.4) are readily accepted by hydrolases. Linalool derivative 4b is alcoholysed stereoselectively by Candida rugosa lipase (E=9.7).@en

Biocatalytic asymmetric reduction of C=C and C=O bonds is highly attractive to produce valuable (chiral) chemicals for the fine and pharmaceutical industry, yet occurs at the expense of reduced nicotinamide adenine dinucleotide coenzyme NADPH that requires recycling. Established methods each have their challenges. Here we developed a light-driven approach based on photosystem I (PSI) by mimicking the natural electron transfer from PSI via ferredoxin (Fd) towards ferredoxin NADP+ reductase (FNR) in vitro. Illumination with red light led to reduction of NADP+ to NADPH with a turnover frequency of 2.55 s−1 (>9000 h−1) at pH 7.5. Light-driven NADPH regeneration by PSI-Fd-FNR was coupled with three oxidoreductases for asymmetric reduction of C=C and C=O bonds, reaching up to 99 % conversion with a turnover number of 3035, and retaining enantioselectivity. This study demonstrates the capacity of a PSI system to drive continuous NADPH-dependent biocatalytic conversions with light.@en

Regioselective aromatic hydroxylation is desirable for the production of valuable compounds. External flavin-containing monooxygenases activate and selectively incorporate an oxygen atom in phenolic compounds through flavin reduction by the nicotinamide adenine dinucleotide coenzyme, and subsequent reaction with molecular oxygen. This study provides the proof of principle of flavoenzyme-catalyzed selective aromatic hydroxylation with coenzyme biomimetics. The carbamoylmethyl-substituted biomimetic in particular affords full conversion in less than two hours for the selective hydroxylation of 5 mM 3- and 4-hydroxybenzoates, displaying similar rates as with NADH, achieving a 10 mM/h enzymatic conversion of the medicinal product gentisate. This biomimetic appears to generate less uncoupling of hydroxylation that typically leads to undesired hydrogen peroxide. Therefore, we show these flavoenzymes have the potential to be applied in combination with biomimetics.@en

Enantioenriched azido alcohols are precursors for valuable chiral aziridines and 1,2-amino alcohols, however their chiral substituted analogues are difficult to access. We established a cascade for the asymmetric azidohydroxylation of styrene derivatives leading to chiral substituted 1,2-azido alcohols via enzymatic asymmetric epoxidation, followed by regioselective azidolysis, affording the azido alcohols with up to two contiguous stereogenic centers. A newly isolated two-component flavoprotein styrene monooxygenase StyA proved to be highly selective for epoxidation with a nicotinamide coenzyme biomimetic as a practical reductant. Coupled with azide as a nucleophile for regioselective ring opening, this chemo-enzymatic cascade produced highly enantioenriched aromatic α-azido alcohols with up to >99% conversion. A bi-enzymatic counterpart with halohydrin dehalogenase-catalyzed azidolysis afforded the alternative β-azido alcohol isomers with up to 94% diastereomeric excess. We anticipate our biocatalytic cascade to be a starting point for more practical production of these chiral compounds with two-component flavoprotein monooxygenases.@en