Deracemisation and stereoinversion by a nanoconfined bidirectional enzyme cascade: dual control by electrochemistry and selective metal ion activation

纳米限域双向酶级联反应实现消旋化和立体异构化:电化学和选择性金属离子活化的双重控制

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Abstract

The unique ability of the 'electrochemical leaf' (e-Leaf) to drive and control nanoconfined enzyme cascades bidirectionally, while directly monitoring their rate in real-time as electrical current, is exploited to achieve deracemisation and stereoinversion of secondary alcohols using a single electrode in one pot. Two alcohol dehydrogenase enzymes with opposing enantioselectivities, from Thermoanaerobacter ethanolicus (selective for S) and Lactobacillus kefir (selective for R) are driven bidirectionally via coupling to the fast and quasi-reversible interconversion of NADP(+)/NADPH catalysed by ferredoxin NADP(+) reductase - all enzymes being co-entrapped in a nanoporous indium tin oxide electrode. Activity of the Lactobacillus kefir enzyme depends on the binding of a non-catalytic Mg(2+), allowing it to be switched off after an oxidative half-cycle, by adding EDTA - the S-selective enzyme, with a tightly-bound Zn(2+), remaining fully active. Racemate → S or R → S conversions are thus achieved in high yield with unprecedented ease.

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