Highly active immobilized hydrogenase from Desulfovibrio gigas

The hydrogenase from the sulfate reducer Desulfovibrio gigas has been immobilized by covalent coupling onto a porous silica support. Two methods have been used: glutaraldehyde activation of aliphatic amino Spherosil and diazotation of aromatic amino Spherosil. The effect of cytochrome C 3 and CC 3 addition during coupling has been investigated. The highest enzymatic activity (4440 U/g support) and immobilization yield (29 %) was obtained when coupling hydrogenase in the presence of cytochrome C 3 or CC 3 with diazotized aromatic amino silica. This immobilized hydrogenase preparation which shows a very good resistance to oxygen inactivation seems suitable for hydrogen photoproduction by coupling with illuminated chloroplasts.