Efficient cofactor regeneration is essential for the utilization of many oxidoreductases. Herein, the beta-subunit of formate dehydrogenase 1 from Methylobacterium extorquens AM1 (MeFDH1-beta) was employed as a novel electroactive biocatalyst for electroenzymatic NADH regeneration for the first time. MeFDH1-beta showed much better properties in both catalytic activity (a turnover frequency of 3,600 hr(-1)) and current efficiency (ca. 98%) than a conventional rhodium (III)-based chemical catalyst (M = [Cp*Rh(bpy)H2O](2+), Cp* = C5Me5, bpy = 2,2'-bipyridine) and a traditional biocatalyst (diaphorase). (C) 2016 The Electrochemical Society. All rights reserved.