Sónia Zacarias, Marisela Vélez, Marcos Pita, Antonio L De Lacey, Pedro M Matias, Inês A C Pereira
Journal: Methods in enzymology 2019;613():169-201
PMID: 30509465
The [NiFeSe] hydrogenases are a subgroup of the well-characterized family of [NiFe] hydrogenases, in which a selenocysteine is a ligand to the nickel atom in the binuclear NiFe active site instead of cysteine. These enzymes display very interesting catalytic properties for biological hydrogen production and bioelectrochemical applications: high H production activity, bias for H evolution, low H inhibition, and some degree of O tolerance. Here we describe the methodologies employed to study the [NiFeSe] hydrogenase isolated from the sulfate-reducing bacteria D. vulgaris Hildenborough and the creation of a homologous expression system for production of variant forms of the enzyme.
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