期刊
JOURNAL OF BIOTECHNOLOGY
卷 307, 期 -, 页码 202-207出版社
ELSEVIER
DOI: 10.1016/j.jbiotec.2019.10.016
关键词
Carboxylate reductase; Multi-enzyme cascade; Cell-free ATP recycling; Aldehyde; Polyphosphate
资金
- Federal Ministry of Science, Research and Economy (BMWFW)
- Federal Ministry of Traffic, Innovation and Technology (bmvit)
- Styrian Business Promotion Agency SFG
- Standortagentur Tirol
- Government of Lower Austria
- Business Agency Vienna through the COMET-Funding Program
- Austrian Research Promotion Agency FFG (FEM-Tech) [870897]
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 beta-nicotinamide adenine dinucleotide 2'-phosphate of 100 mu M 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.
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