对在南极磷虾自溶过程中起主要作用的蛋白酶的分离纯化条件和主要生化性质进行了研究。经过30％～70％硫酸铵分级沉淀，DEAE—Sepharose Fast Flow阴离子交换层析，Sephacryl S-200HR凝胶层析后得到电泳纯的蛋白酶，该酶纯化倍数为12．59，产率为43％，分子质量约为28ku；最适pH和最适温度为pH8．0和50℃；该酶pH稳定范围为pH7．0～9．5，并在45℃以下较稳定；苯甲基磺酰氟（PMSF）、大豆胰蛋白酶抑制剂（SBTI）、对甲苯磺酰基赖氨酸氯甲基酮（TLCK）对该酶活性有较强的抑制作用。%A protease that plays a role on autolysis physical and chemical characteristics were studied. The of Antarctic krill was purified from Antarctic krill and its major purified protease was obtained after 30% - 70% ammonium sulfate grading precipitation, DEAE-Sepharose Fast Flow anion-exchange chromatography and Sephacryl S -200 HR chromatography. The protease was confirmed by a single band on SDS-PAGE and was purified by 12.59-fold with the yield of 43%. The relative molecular mass of the enzyme was determined to be about 28 ku. The casein was used as substrate to measure the characteristics of the protease. The optimum pH and temperature of the enzyme were 8.0 and 50℃ , respectively. The enzyme was stable when pH value is in the range of 7.0 to 9.5 and the temperature is blow 45℃ ; PMSF （phenylmethyl sulfonyl fluoride）, SBTI （strontium bismuth titanate） and TLCK （N-α-Tosyl-L-Lysine Chloromethyl Ketone） can obviously inhibit the enzyme.
展开▼
