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NAD+-dependent hydrogenase from the hydrogen oxidizing bacterium Alcaligenes eutrophus Z1. Stabilization against temperature and urea induced inactivation
Authors:V O Popov  A N Ovchinnikov  A M Egorov  I V Berezin
Affiliation:1. Ecology Institute, Qilu University of Technology (Shandong Academy of Sciences), Shandong Provincial Key Laboratory of Applied Microbiology, Jinan, 250103, PR China;2. School of Science and Enterprise and Research Centre, Institute of Technology Carlow, Carlow, 00353, Ireland;3. Rubenstein School of Environment and Natural Resources, University of Vermont, Burlington, 05405, USA;1. Universidade Federal do Rio de Janeiro, Campus de Xerém, Estrada de Xerém, N° 27, Xerém, Duque de Caxias, Rio de Janeiro 25245-390, Brazil;2. Pontifícia Universidade Católica do Rio de Janeiro, Departamento de Engenharia Industrial, Rua Marquês de São Vicente, 225, Rio de Janeiro, RJ, 22451-900, Brazil;3. NanoBusiness Informação e Inovação Ltda, Polo Tecnológico de Xerém, Prédio 32, Av. Nossa Senhora das Graças, 50, Xerém, Duque de Caxias, Rio de Janeiro 25250-020, Brazil;4. Universidade Federal Fluminense, Instituto de Física, Laboratório de difração de raios X(LDRX-UFF), Av. Gal. Milton Tavares de Souza, Campus Praia Vermelha, Niterói, RJ, 24210-346, Brazil;5. Universidade Federal do Rio de Janeiro, Escola de Belas Artes, Av. Pedro Calmon 500, Rio de Janeiro, RJ, 21941-901, Brazil;6. Centro Universitário Augusto Motta, Av. Paris 84, Rio de Janeiro, Rio de Janeiro 21041-020, Brazil;7. Universidade Federal do Rio de Janeiro, Instituto de Química, Av. Athos da Silveira Ramos, 149, Rio de Janeiro, RJ, 21941-611, Brazil;1. Institute of Biochemistry, Vilnius University, Mokslininku Street 12, LT-08662 Vilnius, Lithuania;2. Faculty of Chemistry, Vilnius University, Naugarduko Street 24, LT-03225 Vilnius, Lithuania;1. CNRS, CRPP, UPR 8641, F-33600 Pessac, France;2. Univ. Bordeaux, CRPP, UPR 8641, F-33600 Pessac, France;1. Departamento de Química, Faculdade de Filosofia Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901 Ribeirão Preto, SP, Brazil;2. Departments of Chemistry and Materials Science and Engineering, University of Utah, Salt Lake City, UT 84112, United States
Abstract:Chemical modification of the NAD+-dependent hydrogenase from the hydrogen oxidizing bacterium Alcaligenes eutrophus Z1 results in considerable enzyme stabilization towards urea and temperature induced inactivation. The stabilizing effect was shown to originate from the suppression of hydrogenase tetramer dissociation. The magnitudes of the stabilizing effects (5-fold and more) were in agreement with the values predicted on the basis of the enzyme thermoinactivation mechanism postulated earlier. Hydrophobic interactions are considered to be critical for the stability of the enzyme quaternary structure. Various methods of hydrogenase immobilization were tested. The enzyme was immobilized with a high retention of activity on aminated silochrom via its carboxylic groups.
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