The asymmetric bioreduction of activated alkenes catalyzed by flavin-dependent enoate reductases from the OYE-family represents a powerful method for the production of optically active compounds. For its preparative-scale application, efficient and economic NADH-recycling is crucial. A novel enzyme-coupled NADH-recycling system is proposed based on the concurrent oxidation of a sacrificial sec-alcohol catalyzed by an alcohol dehydrogenase (ADH-A). Due to the highly favorable position of the equilibrium of ene-reduction versus alcohol-oxidation, the cosubstrate is only required in slight excess.
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http://dx.doi.org/10.1002/bit.23078 | DOI Listing |
J Am Chem Soc
July 2024
School of Life Sciences & Center of Novel Biomaterials, The Chinese University of Hong Kong, Shatin, Hong Kong S.A.R. 999077.
NAD(H)-dependent enzymes play a crucial role in the biosynthesis of pharmaceuticals and fine chemicals, but the limited recyclability of the NAD(H) cofactor hinders its more general application. Here, we report the generation of mechano-responsive PEI-modified Cry3Aa protein crystals and their use for NADH recycling over multiple reaction cycles. For demonstration of its practical utility, a complementary Cry3Aa protein particle containing genetically encoded and co-immobilized formate dehydrogenase for NADH regeneration and leucine dehydrogenase for catalyzing the NADH-dependent l--leucine (l--Leu) biosynthesis has been produced.
View Article and Find Full Text PDFAngew Chem Int Ed Engl
March 2023
Heterogeneous Biocatalysis laboratory. Center for Cooperative Research in Biomaterials (CIC biomaGUNE)-Basque Research and Technology Alliance (BRTA), Paseo de Miramón,182., 20014, Donostia-San Sebastián, Spain.
In vitro biosynthetic pathways that condense and reduce molecules through coenzyme A (CoASH) activation demand energy and redox power in the form of ATP and NAD(P)H, respectively. These coenzymes must be orthogonally recycled by ancillary reactions that consume chemicals, electricity, or light, impacting the atom economy and/or the energy consumption of the biosystem. In this work, we have exploited vinyl esters as dual acyl and electron donor substrates to synthesize β-hydroxy acids through a non-decarboxylating Claisen condensation, reduction and hydrolysis stepwise cascade, including a NADH recycling step, catalyzed by a total of 4 enzymes.
View Article and Find Full Text PDFACS Catal
March 2021
Department of Chemistry, Inorganic Chemistry Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QR, United Kingdom.
Deuterium-labeled nicotinamide cofactors such as [4-H]-NADH can be used as mechanistic probes in biological redox processes and offer a route to the synthesis of selectively [H] labeled chemicals biocatalytic reductive deuteration. Atom-efficient routes to the formation and recycling of [4-H]-NADH are therefore highly desirable but require careful design in order to alleviate the requirement for [H]-labeled reducing agents. In this work, we explore a suite of electrode or hydrogen gas driven catalyst systems for the generation of [4-H]-NADH and consider their use for driving reductive deuteration reactions.
View Article and Find Full Text PDFNat Commun
March 2020
Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, South Parks Road, Oxford, OX1 3QR, UK.
Enzymes dependent on nicotinamide cofactors are important components of the expanding range of asymmetric synthetic techniques. New challenges in asymmetric catalysis are arising in the field of deuterium labelling, where compounds bearing deuterium (H) atoms at chiral centres are becoming increasingly desirable targets for pharmaceutical and analytical chemists. However, utilisation of NADH-dependent enzymes for H-labelling is not straightforward, owing to difficulties in supplying a suitably isotopically-labelled cofactor ([4-H]-NADH).
View Article and Find Full Text PDFBiotechnol Bioeng
March 2019
Department of Chemical & Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore.
Alcohol dehydrogenase (ADH) and amine dehydrogenase (AmDH)-catalyzed one-pot cascade conversion of an alcohol to an amine provides a simple preparation of chiral amines. To enhance the cofactor recycling in this reaction, we report a new concept of coupling whole-cells with the cell-free system to enable separated intracellular and extracellular cofactor regeneration and recycling. This was demonstrated by the respective biotransformation of racemic 4-phenyl-2-butanol 1a and 1-phenyl-2-propanol 1b to (R)-4-phenylbutan-2-amine 3a and (R)-1-phenylpropan-2-amine 3b.
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