A proton-deuterium exchange study of three types ofDesulfovibrio hydrogenases

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Abstract
Hydrogenases are among the main enzymes involved in bacterial anaerobic corrosion of metals. The study of their mode of action is important for a full comprehension of this phenomenon. The three types ofDesulfovibrio hydrogenases [(Fe), (NiFe), (NiFeSe)] present different patterns in the pH dependence of their activity. The periplasmic enzyme fromDesulfovibrio salexigens and the cytoplasmic enzyme fromDesulfovibrio baculatus both have pH optima at 7.5 for H2 uptake and 4.0 for H2 evolution and H+−D2 exchange reaction (measured by membrane-inlet mass-spectrometry). The H2 to HD ratio at pH above 5.0 is higher than 1.0. The periplasmic hydrogenase fromD. gigas presents the same pH optimum (8.0) for the H+−D2 exchange as for H2 consumption. In contrast, the enzyme fromD. vulgaris has the highest activity in H2 production and in the exchange at pH 5.0. Both hydrogenases have a H2-to-HD ratio below 1.0.