Characterization of the active site of a hydrogen sensor from Alcaligenes eutrophus

A third hydrogenase was recently identified in the proteobacterium Alcaligenes eutrophus as a constituent of a novel H-2-sensing multicomponent regulatory system. This regulatory hydrogenase (RH) has been overexpressed in cells deficient in both the NAD(+)-reducing [NiFe]-hydrogenase and the membrane-bound [NiFe]-hydrogenase. EPR, FTIR and activity studies of membrane-free extracts revealed that the RH has an active site much like that of standard [NiFe]-hydrogenases, i.e. a Ni-Fe site with two CN- groups and one CO molecule. Its catalytic power is low, but the RH is always active, insensitive to oxygen, and occurs in only two redox states. (C) 1998 Federation of European Biochemical Societies.