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Towards electroenzymatic processes involving old yellow enzymes and mediated cofactor regeneration

engineering in life science. 2017; 
Andreas Tosstorff Cora Kroner Diederik J. Opperman Frank Hollmann Dirk Holtmann
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Gene Synthesis TsOYE (NCBI accession no. AM902709.1) was synthesized and cloned into pET28b(+) via the NdeI and EcoRI restriction sites (Genscript). Protein expression was performed in Escherichia coli BL21(DE3). Transformed strains were grown in LB medium (10 g/L peptone, 5 g/L yeast extract, and 10 g/L NaCl) containing 35 mg/L kanamycin to an OD600 of approximately 0.8–1.0 (30°C, 200 rpm). Protein expression was induced by adding IPTG to a final concentration of 1 mM.  Get A Quote

摘要

Old yellow enzymes are able to catalyze asymmetric C=C reductions. A mediated electroenzymatic process to regenerate the NADPH in combination with an old yellow enzyme was investigated. Due to the fact that the overall process was affected by a broad set of parameters, a design of experiments (DoE) approach was chosen to identify suitable process conditions. Process conditions with high productivities of up to 2.27 mM/h in combination with approximately 90% electron transfer efficiency were identified.

关键词

C=C reduction Design of experiments Electroenzymatic synthesis Mediated electron transfer Old yellow enzyme
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