汉麻球蛋白酶解产物中巯基肽的含量及分布研究  

The Content and Distribution of Mercapto Peptides in the Enzymatic Hydrolysates of of Hemp Globulin

作　　者：石杰[1] 魏连会[1] 张正海[1] 宋淑敏[1] 杨庆丽[1] 董艳[1] Shi Jie;Wei Lianhui;Zhang Zhenghai;Song Shumin;Yang Qingli;Dong Yan(Daqing Branch of Heilongjiang Academy of Sciences,Daqing 163319)

机构地区：[1]黑龙江省科学院大庆分院,大庆163319

出　　处：《中国粮油学报》2023年第3期109-114,共6页Journal of the Chinese Cereals and Oils Association

基　　金：黑龙江省科学院科学研究基金(KY2022DQ01);大庆市指导性计划项目(zd-2021-93)。

摘　　要：采用DTT还原结合Sephadex LH-20葡聚糖凝胶层析的方法分离测定了汉麻球蛋白3种酶解产物中巯基肽的含量,测定结果显示:碱性蛋白酶>胃蛋白酶>木瓜蛋白酶。采用LC-MS-MS检测并鉴定出3种蛋白酶酶解物中巯基肽的分布和序列,结果显示:碱性蛋白酶酶解物中巯基肽数量为41个,木瓜蛋白酶和胃蛋白酶酶解物中都是31个,来源均是Edestin2、Edestin3、清蛋白和非特征蛋白。3种酶酶解获得的麻仁球蛋白(Edestin)巯基肽大部分为亲水性的,分子质量主要分布于1000~2000 u,其中10个是3种酶解产物共有的,碱性蛋白酶和胃蛋白酶酶解物中有4个相同的来源于Edestin3的麻仁球蛋白巯基肽。胃蛋白酶酶解得到1个独有的源于Edestin2的麻仁球蛋白巯基肽。碱性蛋白酶酶解得到了4个独有的源于Edestin3的麻仁球蛋白巯基肽。3种蛋白酶由于作用位点和效力不同得到的麻仁球蛋白巯基肽数量和分布均有不同,但是碱性蛋白酶酶解汉麻球蛋白可以获得更多的麻仁球蛋白巯基肽,推测碱性蛋白酶可以更好地打开汉麻球蛋白中的二级及以上结构,使二硫键更多地暴露出来。The content of mercapto peptides in three kinds of enzymatic hydrolysates of hemp globulin was separated and determined by DTT reduction combined with Sephadex LH-20 sephadex del chromatography.The results showed that:alkaline protease>pepsin>papain.The distribution and sequence of mercapto peptides in the three kinds of protease hydrolysates were detected and identified by LC-MS-MS.The results showed that there were 41 mercapto peptides in alkaline protease hydrolysates,31 mercapto peptides in papain and pepsin hydrolysates,and the sources were Edestin2,Edestin3,albumin and uncharacterized proteins.Most of the mercapto peptides of Edestin obtained by the three enzymes were hydrophilic,and their molecular weights were mainly distributed in the range of 1000~2000 u,of which ten were common to the three enzymatic hydrolysates.There were four identical Edestin3-derived mercapto peptides in the alkaline protease and pepsin hydrolysates.A unique mercapto peptide from Edestin2 was obtained by pepsin digestion.Four unique mercapto peptides derived from Edestin3 were obtained by alkaline protease hydrolysis.The number and distribution of mercapto peptides of edestin obtained by the three proteases were different due to their different action sites and effects.However,more mercapto peptides of hemp globulins could be obtained by enzymatic hydrolysis of hemp globulins with alkaline proteases.It was speculated that alkaline protease could better open the secondary and higher structures of hemp globulins and expose more disulfide bonds.

关 键 词：汉麻球蛋白 酶解 麻仁球蛋白 巯基肽 分布 

分 类 号：TS229[轻工技术与工程—粮食、油脂及植物蛋白工程]