章鱼内脏胰蛋白酶抑制剂的分离纯化与性质研究-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Purification and Characterization of a Trypsin Inhibitor from the Viscera of Octopus(Octopus fangsiao)

Author(s):: WU Hai-longCHEN Xue-fang; WANG Cheng; CAI Qiu-feng; CAO Min -jie; (College of Biological Engineering,Jimei University,Xiamen 361021, China)

摘要:: 通过热处理、硫酸铵盐析、膜超滤及SP-Sepharose 离子交换层析，从章鱼内脏中纯化得到一种胰蛋白酶抑制剂 (Octopus Trypsin Inhibitor ，OTI)．Tricine-SDS-PAGE 电泳结果显示，OTI的分子质量约为6500 u．性质分析结果表明：OTI在pH=1.0~11.0缓冲液中孵育30 min，抑制活性稳定；在100 ℃下加热1 h、2 h 后仍分别具有90% 和65% 的抑制活性，具有良好的pH值稳定性和热稳定性；OTI能够显著抑制胰蛋白酶的活性，对胰凝乳蛋白酶、胃蛋白酶和木瓜蛋白酶几乎无抑制活性；OTI能够有效抑制蓝圆鲹肌肉中肌原纤维结合型丝氨酸蛋白酶 (MBSP) 引起的肌球蛋白降解．

Abstract:: An octopus trypsin inhibitor (OTI) was purified from the viscera of octopus by processes including heat treatment ，ammonium sulfate fractionation，ultrafiltration and SP- Sepharose ion exchange chromatography.The purified OTI was in homogeneity with a molecular mass of approximately 6500 u on Tricine-SDS-PAGE.OTI maintained high activity at pH range of 1.0~11.0.More than 90% and 65% of its initial inhibitory activity remained after incubation at 100 ℃ for 1 h and 2 h ，respectively，suggesting its high stability under high temperature.OTI was specifically active against trypsin，while revealed no inhibitory activity to pepsin，chymotrypsin and papain.Furthermore，OTI was effective in suppressing the degradation of myofibrillar proteins in the skeletal muscle of blue scad ( Decapterus maruadsi) caused by an endogenous myofibril-bound serine proteinase (MBSP).