Purification of Hydrogenase by Fast Protein Liquid Chromatography and by Conventional Separation Techniques: A Comparative Study

Abstract
Hydrogenase was purified from the photosynthetic bacterium Thiocapsa roseopersicina to homogeneity by various methods. Conventional techniques included separation of the crude protein extract on Phenyl-Sepharose CL 4B, DEAE-cellulose DE52, and chromatofocusing columns or on preparative polyacrylamide gelelectrophoresis. The same protein was isolated by fast protein liquid chromatography (FPLC) in two steps. Comparison of the two different approaches clearly show the superiority of the FPLC method both in enzyme recovery yield and in time requirement.

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