3942中性蛋白酶酶解中国毛虾蛋白制备血管紧张素转移酶抑制肽  

Angiotensin I converting enzyme inhibitory peptide from 3942 neutral proteinase digests of Acetes chinensis proteins

作　　者：许萍[1] 薛长湖[1] 张香治[1] 赵雪[1] 李兆杰[1] 

机构地区：[1]中国海洋大学水产学院,山东青岛266003

出　　处：《中国水产科学》2005年第4期501-505,共5页Journal of Fishery Sciences of China

基　　金：国家"863"高技术研究发展项目(2004AA6250100);国家"十五"攻关重大专项资助项目(2001BA501A26).

摘　　要：采用3942中性蛋白酶酶解中国毛虾(Aceteschinensis)蛋白,制备具有抑制血管紧张素转移酶(ACE)活性的多肽。采取正交实验研究料水比、酶量、反应温度、反应时间4个因素对酶解中国毛虾蛋白产物的ACE抑制活性的影响。结果显示,在料水比1∶3(体积比),酶量0.5%,温度45℃,时间8h的水解条件下得到的酶解产物F7的ACE抑制活性最高,其抑制率达到92.86%。用层析柱SephadexG25、SPSephadexC50对F7进行分离纯化,选取ACE抑制活性最高的峰进一步用反相高压液相色谱进行纯化,得到单一组分FⅠ。经飞行时间质谱和碰撞诱导裂解分析确定,FⅠ为新型的ACE抑制肽SerPro,IC50值为272μmol/L。3942 neutral proteinase was used to hydrolyse Acetes chinensis protein to prepare active peptides, which could inhibit the activity of angiotensin I-converting enzyme(ACE). In order to study the influence on ACE inhibitory activity of hydrolysate, four levels of enzyme dosages, material:water (V/V),hydrolysis time,and hydrolysis temperature were changed according to the orthogonal table. The results revealed the optimum condition for active peptide were V_(material)/V_(water)=1/3,enzyme dosage 0.5%, temperature 45?℃, hydrolysis time 8?h. Then F7-4 was separated and purified by Sephadex G-25 and SP-Sephadex C-50. The most potent fraction was concentrated and further purified by preparative reverse-phase high-performance liquid chromatography (HPLC) to get pure sample FⅠ. FⅠ was identified as Ser-Pro,of which the IC_(50) value was 296?μmol/L,after the application of time-of-flight mass spectrometry (TOFMS) and high-energy collision-induced dissociation (CID).

关 键 词：中国毛虾 血管紧张素转移酶抑制肽 3942中性蛋白酶 

分 类 号：TS254.9[轻工技术与工程—水产品加工及贮藏工程]