Hydrogenases are a class of enzymes responsible for hydrogen gas production in many microorganisms. Their study is considered of great interest for biotechnological applications to support a fossil fuel-free world. To this end, the construction of artificial devices that are able to use solar energy to produce hydrogen (the so-called “nano-leaf”) has been proposed. Because of its relatively simple structure and high specific activity, Chlamydomonas reinhardtii HydA1 hydrogenase (CrHydA1) is an enzyme of outstanding interest for this purpose. Thus, the aim of this study is to produce and characterize CrHydA1.
Since CrHydA1 is quickly inactivated by oxygen, this study required the development of protocols for CrHydA1 production, manipulation and storage in strictly anoxic conditions. The recombinant expression system in Escherichia coli developed at the National Renewable Energy Laboratory (NREL), USA, was used to produce and purify CrHydA1 to homogeneity with a specific activity of abo ...