[Stability of the hydrogenase from Tetraselmis subcordiformis and its preliminary purification].
Tetraselmis subcordiformis, a marine green alga, can produce hydrogen by photobiologically hydrolyzing seawater with hydrogenase. In this study, the preliminary purification of the enzyme was explored by ammonium sulfate precipitation, and the impact of sodium dithionite, beta-mercaptoethanol and glycerol on the enzyme stability during the process was investigated. The experimental results illustrated that sodium dithionite provided significant protection on the hydrogenase by depleting oxygen, while glycerol, a protectant against the structure instability of the enzyme, also presented protection. Crude enzyme with specific activity of 0.557 U/mg protein was extracted using 60%-70% saturated ammonium sulfate solution supplemented with 200 mmol/L sodium dithionite and 5% glycerol, and the hydrogenase recovery yield was about 30%.
Yan
Fei
F
Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy ofSciences, Dalian 116023, China.
Chen
Zhao'an
Z
Cao
Xupeng
X
Lu
Hongbin
H
Xue
Song
S
Zhang
Wei
W
D000645:Ammonium Sulfate / Q000737:chemistry; D011232:Chemical Precipitation; D000460:Chlorophyta / Q000201:enzymology*; D004795:Enzyme Stability; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D012623:Seawater
D006859:Hydrogen; D006864:Hydrogenase; D000645:Ammonium Sulfate