Characterization of the [NiFeSe] hydrogenase from Desulfovibrio vulgaris Hildenborough.

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

Abstract

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.

© 2018 Elsevier Inc. All rights reserved.

Address: Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.; Instituto de Catálisis y Petroleoquímica, CSIC, Madrid, Spain.; Instituto de Catálisis y Petroleoquímica, CSIC, Madrid, Spain. Electronic address: [email protected].; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal; iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal. Electronic address: [email protected].; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal. Electronic address: [email protected].

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