| 摘要 |
PURPOSE: To obtain the subject highly thermally stable new enzyme having high optimal temperature, useful for reagents or bioreactors, by culturing microbes belonging to the genus Thermococcus and having heat-resistant ATPase productivity. CONSTITUTION: Microbes belonging to the genus Thermococcus and having ATPase productivity [e.g. Thermococcus K1 strain (FERM P-14622) ] is subjected to stationary culture anaerobically in a medium at 90 deg.C for 8hr followed by centrifugation to collect the resultant microbes, which are then crushed in a buffer solution followed by cetrifugation to separate a membrane fraction. The membrane fraction is put to extraction with a surfactant-contg. buffer solution, and the resultant extract is purified by column chromatography, thus obtaining the objective new heat-resistant ATPase having the following characteristics: hydrolyzing nucleotide triphosphate into nucleotide diphosphate and inorganic phosphoric acid; requiring Mg<2+> , Mn<2+> , Co<2+> , Ni<2+> , Zn<2+> , Ca<2+> as cofactor; rising activity by HSO<-> 3 , S2 O<2-> 4 , HCO<-> 3 ; optimal pH: 6-6.5; optimal temperature; 90-95 deg.C; molecular weight: 600kDa. |
