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Purification of Hydrogenase from Thiocapsa roseopersicina : Effect of Ammonium Sulfate Precipitation and Heat-Treatment  

Choi, Eun-Hye (Bioenergy Research Center, Korea Institute of Energy Research)
Oh, You-Kwan (Bioenergy Research Center, Korea Institute of Energy Research)
Kim, Mi-Sun (Bioenergy Research Center, Korea Institute of Energy Research)
Publication Information
Transactions of the Korean hydrogen and new energy society / v.17, no.4, 2006 , pp. 371-378 More about this Journal
Abstract
Effect of $(NH_4)_2SO_4$ precipitation and heat-treatment on hydrogenase which was extracted from the cytoplasmic fraction of the phototrophic purple sulfur bacterium Thiocapsa roseopersicina NCIB 8347 was studied. Crude enzyme extract was prepared by centrifugation($28,000{\times}g$, $400,000{\times}g$) after sonication of cells grown under photosynthetic condition for 96 hrs. Various conditions of $(NH_4)_2SO_4$ precipitation and heat-treatment were examined and the effect of protein concentration was analyzed by SDS-electrophoresis between the treatments. Optimum conditions for $(NH_4)_2SO_4$ precipitation and heat-treatment for evolution hydrogenase activity were 40-60% saturation and $60^{\circ}C$ for 20 min, respectively, which exhibited the specific hydrogenase activity of 0.78 U/mg-protein. Specific hydrogenase activity was decreased to 31.6% when the heat-treatment at $60^{\circ}C$ increased from 20 min to 5 hrs.
Keywords
thiocapsa roseopersicina$(NH_4)_2SO_4$ precipitation; heat-treatment; hydrogenase;
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Times Cited By KSCI : 1  (Citation Analysis)
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