食品科学 ›› 2012, Vol. 33 ›› Issue (7): 168-171.doi: 10.7506/spkx1002-6630-201207036

• 生物工程 • 上一篇    下一篇

章鱼消化道蛋白酶的分离纯化及性质

任 佩1,王 莹1,金玉兰2,朴美子1,   

  1. 1.青岛农业大学食品科学与工程学院 2.青岛农业大学化学与药学院
  • 出版日期:2012-04-15 发布日期:2012-04-20

Extraction, Purification and Characterization of Protease from Digestive Tract of Octopus vulgaris

REN Pei1,WANG Ying1,JIN Yu-lan2,PIAO Mei-zi1,*   

  1. (1. College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China; 2. College of Chemistry and Pharmaceutical Sciences, Qingdao Agricultural University, Qingdao 266109, China)
  • Online:2012-04-15 Published:2012-04-20

摘要: 以章鱼加工下脚料消化道为原料,经硫酸铵沉淀、纤维素CM-52阳离子交换层析、DEAE-Sephadex A50阴离子交换层析、SDS-PAGE电泳等方法,从中提取纯化出一种蛋白酶电泳纯样品OP-I,并对其性质进行研究。结果表明:该酶分子质量为80.5kD。最适反应温度为55~60℃,pH值为7~9。金属蛋白酶抑制剂(EDTA)可以完全抑制该酶的活性。Mn2+、Ca2+、Mg2+对OP-I有激活作用,酶促动力学研究显示其米氏常数Km为0.33mmol/L,Vmax为66.7mg/(mL ·min)。

关键词: 章鱼, 蛋白酶, 纯化, 性质

Abstract: An electrophoretically pure protease , named as OP-I, was obtained from the digestive tract of Octopus vulgaris by ammonium sulfate precipitation, cellulose CM-52 cation-exchange chromatography, DEAE-Sephadex A50 anion-exchange chromatography and SDS-PAGE. Its properties were also characterized. The results showed that the molecular weight of the protease was 80.5 kD and its optimal reaction temperature and pH were 55-60 ℃ and 7-9, respectively. OP-I could be completely inhibited by EDTA, a metalloproteinase inhibitor. In contrast, Mn2+, Ca2+ and Mg2+could stimulate OP-I activity. Moreover, the Km of OP-I was 0.33 mmol/L and Vmax was 66.7 mg/(mL ·min)as determined by kinetic studies.

Key words: Octopus vulgaris, protease, purification, characterization

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