海洋科学2016，Vol.40Issue(10)：97-104,8.DOI:10.11759/hykx20151224003
三疣梭子蟹(Portunus trituberculatus)肝胰腺内源酶性质的初步研究
Characteristics of endogenous protease from hepatopancreas of swimming crab (Portunus trituberculatus)
摘要
Abstract
Endogenous autolytic enzyme is an important enzyme in Portunus trituberculatus. Normally, after the death of swimming crabs, deterioration is very rapid in the muscle, which produces a severe effect on the storage quality of crabs. To investigate few basic properties of this autolytic enzyme, crude samples were extracted from the hepatopancreas of P. trituberculatus and the parameters are optimized. Then, a preliminary study on the enzymatic properties of the endogenous enzyme was conducted. The results showed that the optimum extraction time range of endogenous protease was 4–12 h, and the specific activity was significantly higher than that at other time points,i.e., 0–4 h and 12–14 h. The optimal temperature and pH for hydrolysis of casein were 65℃ and 7.0–8.0, respectively. Compared with other protease inhibitors, endogenous protease activity was significantly inhibited by serine prote-ase inhibitors, including soybean trypsin inhibitor (SBTI) and phenylmethylsulfonyl fluoride (PMSF), suggesting that serine protease was the dominant content of this endogenous enzyme. Ammonium sulfate grading precipitation showed that fractions of 0%–70% and 30%–70% could be used to collect the targeted proteases for casein and Boc-Phe-Ser-Arg-MCA hydrolysis, respectively. Their specific activities were significantly higher than those of the samples precipitated by other ammonium sulfate fractions. Moreover, samples collected by ammonium sulfate frac-tions of 40%–60% had a higher protein content and total activity of crude and serine proteinase than those of the samples precipitated by other ammonium sulfate fractions.关键词
三疣梭子蟹(Portunustri tuberculatus)/肝胰腺/内源酶/性质Key words
Swimming crab (Portunus trituberculatus)/Hepatopancreas/Endogenous protease/Characteristic分类
轻工纺织引用本文复制引用
杨利珠,张莉,黄琳,孟祥红..三疣梭子蟹(Portunus trituberculatus)肝胰腺内源酶性质的初步研究[J].海洋科学,2016,40(10):97-104,8.基金项目
山东省自然科学基金项目(ZR2015CM010)Province Natural Science Foundation of Shandong, No.ZR2015CM010 （ZR2015CM010）
