| Popis: |
Publisher Summary This chapter focuses on the physical and chemical properties of purified hydrogenase, its possible mechanism of action, and its role in energy metabolism. The techniques used for the purification of various hydrogenases are similar to those commonly used for protein purification. The major requirement for purification of hydrogenase in good yield is the complete absence of oxygen. It makes no difference what assay for hydrogenase is used during purification, provided that when the assay is coupled to further reduction, such as NAD+ reduction, hydrogenase is limiting and not the oxidant. Many sulfhydryl reagents inhibit hydrogenase, and because of this, it has been suggested by several investigators that sulfhydryl groups are involved in the activity of the enzyme. The main approaches that have been used in attempts to understand the mechanism of catalysis by hydrogenase have been kinetic, H/D exchange, and para-ortho conversion studies and chemical and physical studies of the structure of the pure enzyme. |
