Separation and purification of proteinases from Gastric Mucosa of Northern Sheatfish,Silurus soldatovi  

作　　者：刘伟 张秀梅 战培荣 张照韩 

机构地区：[1]Heilongjiang River Fisheries Research Institute,Chinese Academy of Fishery Sciences [2]College of Life Science and Technology,Ocean University of China [3]State Key Laboratory of Urban Water Resource and Environment,Harbin Institute of Technology

出　　处：《Journal of Harbin Institute of Technology(New Series)》2011年第3期62-67,共6页哈尔滨工业大学学报（英文版）

基　　金：Sponsored by the Key Research Program of Chinese Academy of Fishery Sciences (Grant No. 1999-01-10);Agricultural Science and Technology Fund of Department of Science and Technology (Grant No. 02EFN216900727);the National Natural Science Foundation of China (Grant No. 50638020)

摘　　要：The separation and purification method for pepsin of Northern Sheatfish (Silurus soldatovi) was established by using the combination technology of salting-out,gel chromatography and gel electrophoresis,and the enzymic properties were also analyzed.The experimental results indicated that 28% and 56% (NH4)2SO4 saturation could separate the activated protease from the pepsin extract of gastric mucosa of Sheatfish (Silurus soldatovi) ;compared with the homogenate extraction,the pepsin specific activity of purified extraction by Sephadex G-75 gel chromatography system increased 598 fold,was 5 times higher than that of activated liquid,and the total production rate was 10.1%.The purified pepsin liquor at the conditions of pH3.3 0.01 M alanine-formic acid buffering solution,60 cm chromatography column,and the flowing rate of 0.8 ML/min was analyzed by SDS-PAGE,which indicated that there were two bands and the molecular weight was 34.0 kDa and 40.4 kDa,respectively.There were two peaks in the enzyme activity determination of the separated collecting liquor in gel chromatography,and the SDS-PAGE showed the concentrations of the two proteins was different,which indicated that it existed at least two pepsins in the gastric mucosa of Sheatfish (Silurus soldatovi).The separation and purification method for pepsin of Northern Sheatfish （Silurus soldatovi） was established by using the combination technology of salting-out, gel chromatography and gel electrophoresis, and the enzymic properties were also analyzed. The experimental results indicated that 28 % and 56% （ NH4 ）2 SO4 saturation could separate the activated protease from the pepsin extract of gastric mucosa of Sheatfish （ Silurus soldatovi） ; compared with the homogenate extraction, the pepsin specific activity of purified extraction by Sephadex G--75 gel chromatography system increased 598 fold, was 5 times higher than that of activated liquid, and the total production rate was 10. 1%. The purified pepsin liquor at the conditions of pH3.3 0. 01 M alanine-formic acid buffering solution, 60 cm chromatography column, and the flowing rate of 0. 8 ML/min was analyzed by SDS-PAGE, which indicated that there were two bands and the molecular weight was 34. 0 kDa and 40. 4 kDa, respectively. There were two peaks in the enzyme activity determination of the separated collecting liquor in gel chromatography, and the SDS-PAGE showed the concentrations of the two proteins was different, which indicated that it existed at least two pepsins in the gastric mucosa of Sheatfish （Silurus soldatovi）.

关 键 词：Sheatfish (Silurus soldatovi) PEPSIN separation and purification PROPERTY 

分 类 号：Q949.288.4[生物学—植物学]