以水解试验为导向,采用硫酸铵分级沉淀、Sephadex G-100凝胶层析、DEAE·DE52 阴离子交换柱层析等手段分离纯化南美白对虾虾头主要自溶酶,同时采用抑制剂试验及SDS-PAGE对纯化蛋白酶进行鉴定.结果表明,南美白对虾虾头主要自溶酶经过纯化后比活力高达2.2×106U/g,纯化倍数为96.90.丝氨酸蛋白酶抑制剂苯甲基磺酰氟(PMSF)对主要自溶酶的抑制率高达91.3%;SDS-PAGE显示该酶为单一电泳带,且在电泳中与标准胰蛋白酶移动距离相同,表明南美白对虾虾头内的主要自溶酶为胰蛋白酶.%With hydrolysis test as an orientation, the major autoenzyme was purified from the head of white shrimp (Penneus vannamei) by sulfate precipitation, gel filtration on Sephadex G-100 followed by ion-exchange chromatography DEAE-DE52. The autoenzyme was then identified by inhibitor tests and SDS-PAGE analysis. The results showed the major autoenzyme from the head of white shrimp had a specific activity of 2.2 × 106 U/g with a 96.90-fold increase over that of the crude enzyme extract. This autoenzyme was inhibited by serine protease inhibitor phenyl methyl sulfonyl fluoride (PMSF) with the inhibition rate being 91.3％ , and illustrated as a single band in SDS-PAGE with the same distance as that of standard trypsin. These results indicated that trypsin was the major autoenzyme in the head of white shrimp.
展开▼
