Synthetic Biomimetic Coenzymes and Alcohol Dehydrogenases for Asymmetric Catalysis

Laia Josa-Culleré,Francesca Paradisi,Frank Hollmann,Diederik Opperman,Jian-He Xu,Serena Gargiulo,Aubert Ribaucourt,Antti Lahdenperä,Caroline Paul,Georg Höfler,Jennifer Cassidy,Yuan-Yang Liu

doi:10.3390/catal9030207

Abstract

Redox reactions catalyzed by highly selective nicotinamide-dependent oxidoreductases are rising to prominence in industry. The cost of nicotinamide adenine dinucleotide coenzymes has led to the use of well-established elaborate regeneration systems and more recently alternative synthetic biomimetic cofactors. These biomimetics are highly attractive to use with ketoreductases for asymmetric catalysis. In this work, we show that the commonly studied cofactor analogue 1-benzyl-1,4-dihydronicotinamide (BNAH) can be used with alcohol dehydrogenases (ADHs) under certain conditions. First, we carried out the rhodium-catalyzed recycling of BNAH with horse liver ADH (HLADH), observing enantioenriched product only with unpurified enzyme. Then, a series of cell-free extracts and purified ketoreductases were screened with BNAH. The use of unpurified enzyme led to product formation, whereas upon dialysis or further purification no product was observed. Several other biomimetics were screened with various ADHs and showed no or very low activity, but also no inhibition. BNAH as a hydride source was shown to directly reduce nicotinamide adenine dinucleotide (NAD) to NADH. A formate dehydrogenase could also mediate the reduction of NAD from BNAH. BNAH was established to show no or very low activity with ADHs and could be used as a hydride donor to recycle NADH.

Highlights

The field of biocatalysis has flourished in the past few decades, with enzymes increasingly being used in industrial processes for fine chemical commodities
Enzyme recognition of nicotinamide adenine dinucleotide (NAD)(P), usually by a cofactor motif such as the Rossmann fold, Enzymefor recognition of NAD(P), usually by a cofactor motif such the Rossmann fold, is important cellular processes, when using in vitro systems, the as adenine dinucleotide is important for cellular processes, when using in vitro systems, the adenine dinucleotide moiety of the cofactor becomes obsolete and disadvantageous, being prone to hydrolysis [4]
Due to the lack of activity with purified enzyme, we investigated the use of BNAH as a hydride

Summary

Introduction

The field of biocatalysis has flourished in the past few decades, with enzymes increasingly being used in industrial processes for fine chemical commodities. Catalysts 2019, 9 FOR PEER REVIEW or aldehydes to the corresponding (enantioenriched) alcohols through the use of one equivalent of or aldehydes to the corresponding through of one equivalent of nicotinamide adenine dinucleotide(enantioenriched) cofactor NAD(P).alcohols. ADHs are knownthe to use be specific to either the nicotinamide adenine dinucleotide cofactor NAD(P). ADHs are known to be specific to either the phosphorylated NADP or non-phosphorylated NAD, a few have been found to accept phosphorylated or non-phosphorylated. Enzyme recognition of NAD(P), usually by a cofactor (binding) motif such as the Rossmann fold, Enzymefor recognition of NAD(P), usually by a cofactor (binding) motif such the Rossmann fold, is important cellular processes, when using in vitro systems, the as adenine dinucleotide is important for cellular processes, when using in vitro systems, the adenine dinucleotide moiety of the cofactor becomes obsolete and disadvantageous, being prone to hydrolysis [4]

Methods

Results

Conclusion