Co-factor demand and regeneration in the enzymatic one-step reduction of carboxylates to aldehydes in cell-free systems

Gernot Strohmeier, Anna Schwarz, Jennifer N. Andexer, Margit Winkler

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Addressing the challenges associated with the development of in vitro biocatalytic carboxylate reductions for potential applications, important aspects of the co-factor regeneration systems and strategies for minimizing over-reduction were investigated. The ATP recycling can be performed with similarly high efficiency exploiting the polyphosphate source by combining Meiothermus ruber polyphosphate kinase and adenylate kinase or with Sinorhizobium meliloti polyphosphate kinase instead of the latter. Carboxylate reductions with the enzyme candidates used in this work allow operating at co-factor concentrations of adenosine 5'-triphosphate and β-nicotinamide adenine dinucleotide 2'-phosphate of 100 µM and, thereby, reducing the amounts of alcohols formed by side activities in the enzyme preparations. This study confirmed the expected benefits of carboxylic acid reductases in chemoselectively reducing the carboxylates to the corresponding aldehydes while leaving reductively-sensitive nitro, ester and cyano groups intact.
Original languageEnglish
Number of pages14
JournalJournal of Biotechnology
DOIs
Publication statusAccepted/In press - 28 Oct 2019

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Cell-Free System
Aldehydes
Regeneration
Adenosine Triphosphate
Sinorhizobium meliloti
Adenylate Kinase
Polyphosphates
Recycling
Enzymes
NADP
Esters
Adenosinetriphosphate
Alcohols
Carboxylic acids
NAD
Adenosine
Phosphates
polyphosphate kinase
In Vitro Techniques
carboxylic acid reductase

Fields of Expertise

  • Human- & Biotechnology

Cite this

Co-factor demand and regeneration in the enzymatic one-step reduction of carboxylates to aldehydes in cell-free systems. / Strohmeier, Gernot; Schwarz, Anna; Andexer, Jennifer N.; Winkler, Margit.

In: Journal of Biotechnology, 28.10.2019.

Research output: Contribution to journalArticleResearchpeer-review

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