单秀民.葵花籽粗油体中内源性蛋白酶系的性质探究以及鉴定[J].中国油脂,2020,45(11):68~72.[SHAN Xiumin.Properties and identification of endogenous proteases in crude sunflower seed oil body[J].China Oils and Fats,2020,45(11):68~72.]
葵花籽粗油体中内源性蛋白酶系的性质探究以及鉴定
Properties and identification of endogenous proteases in crude sunflower seed oil body
  
DOI:10.12166/j.zgyz.1003-7969/2020.11.014
中文关键词:  葵花籽;粗油体;内源性蛋白酶系
英文关键词:sunflower seed; crude oil body; endogenous protease
基金项目:国家自然科学青年基金(31301496)
作者单位
单秀民 江南大学 食品学院,江苏 无锡 214122 
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中文摘要:
      通过电泳探究葵花籽粗油体中内源性蛋白酶系的水解性质,以及用液相色谱-串联质谱(LC-MS/MS)鉴定内源性蛋白酶的种类。结果表明,油质蛋白在pH 4~8水解速度相对较快,22 kDa球蛋白在pH 3~5水解速度相对较快,35 kDa球蛋白在pH 4~8水解速度相对较快。葵花籽粗油体中内源性蛋白酶系在pH 4、50 ℃水解速度相对最快。通过抑制剂实验发现,在pH 4下,葵花籽粗油体中有金属蛋白酶类、天冬氨酸蛋白酶类、丝氨酸蛋白酶类的酶活表征,几乎无半胱氨酸蛋白酶酶活的存在。通过LC-MS/MS分析发现,葵花籽粗油体中存在金属蛋白酶类、天冬氨酸蛋白酶类和丝氨酸蛋白酶类和天冬氨酸蛋白酶类抑制剂、丝氨酸蛋白酶类抑制剂以及半胱氨酸蛋白酶类抑制剂。研究结果为利用内源性蛋白酶类进行水相加工提供了理论基础。
英文摘要:
      The hydrolytic properties of endogenous proteases in crude sunflower seed oil body were analyzed by Tricine-SDS-PAGE, and the types of endogenous proteases were identified by LC-MS/MS. The results showed that the hydrolysis rate of oleosins was relatively faster at pH 4-8. 22 kDa globulin was rapidly hydrolyzed at pH 3-5, and 35 kDa globulin was rapidly hydrolyzed at pH 4-8. The hydrolysis rates of endogenous proteases in crude sunflower seed oil body were relatively the fastest at pH 4 and 50 ℃. The results of inhibitor experiments showed that at pH 4, the activities of metalloproteinases, aspartic protease and serine proteases were characterized in crude sunflower seed oil body, and there was almost no cysteine protease activity. LC-MS/MS analysis revealed that the metalloproteinases, aspartic protease and serine proteases, and aspartic proteases inhibitor, serine proteases inhibitor and cysteine protease inhibitor were presented in crude sunflower seed oil body. The research results provided a theoretical basis for the use of endogenous proteases for aqueous processing.
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