沙棘籽粕中乙醇脱氢酶激活肽的制备工艺  

Preparation Process of Ethanol Dehydrogenase Activating Peptide from Hippophae rhamnoides Linn.Seed Meal

作　　者：王志成 王洪伦[2,3] 胡娜 欧阳健[2,3] 何彦峰[1] WANG Zhicheng;WANG Honglun;HU Na;OUYANG Jian;HE Yanfeng(College of Pharmacy,Qinghai Minzu University,Xining 810007;Key Laboratory of Tibetan Medicine Research,Northwest Institute of Plateau Biology,Chinese Academy of Science,Xining 810008;Huzhou China-Science Innovation Center of Plateau Biology,Huzhou 313000)

机构地区：[1]青海民族大学药学院,西宁810007 [2]中国科学院藏药研究重点实验室青海省藏药研究重点实验室中国科学院西北高原生物研究所,西宁810008 [3]湖州中科高原生物创新中心,湖州313000

出　　处：《食品工业》2024年第4期101-108,共8页The Food Industry

基　　金：中国科学院王宽诚率先人才计划“产研人才扶持项目”。

摘　　要：以沙棘籽粕为原料,优化乙醇脱氢酶激活肽的制备工艺并进行表征。通过碱提酸沉和过氧化氢脱色的方法提取纯化沙棘籽粕蛋白,并以乙醇脱氢酶(ADH)激活率和蛋白质回收率为指标,利用单因素试验和响应面法优化沙棘籽粕ADH激活肽的酶解制备工艺,并对所获得肽进行表征。结果表明:沙棘籽粕ADH激活肽的最优酶解工艺参数为pH 10.7、温度54.8℃、酶解时间4.2 h、碱性蛋白酶添加量0.3%、底物浓度5%。在此条件下,沙棘籽粕酶解肽的实际ADH激活率为82.40%,蛋白质回收率为90.81%。氨基酸组成分析结果表明,沙棘蛋白和酶解肽中含量相对较高的是谷氨酸、精氨酸和天冬氨酸。SDS-PAGE结果显示,沙棘蛋白能被碱性蛋白酶较好地水解,酶解肽分子量集中在15 kDa以下。此研究可为沙棘植物资源在食品和保健品方面的进一步开发利用提供一定的理论基础和科学依据。The preparation process was optimized and further characterized of ethanol dehydrogenase activating peptide from Hippophae rhamnoides Linn.(sea buckthorn)seed meal.The sea buckthorn seed meal protein was achieved through alkaline extraction,acid precipitation.and hydrogen peroxide decolorization.The activation rate of ethanol dehydrogenase(ADH)and protein recovery were employed as indicators to optimize the enzymatic hydrolysis preparation process of peptides from sea buckthorn by single factor experiments and response surface methodology.Then,the obtained peptides were characterized.The optimal enzymatic hydrolysis process parameters were as following:pH 10.7,temperature 54.8℃,enzymatic hydrolysis time 4.2 h,alkaline protease addition 0.3%,and substrate concentration 5%.Under the optimal conditions,the ADH activation rate of enzymatic hydrolysis peptides could achieve 82.40%,and the protein recovery rate could reach 90.81%.The analysis of amino acid composition showed that glutamic acid,arginine,and aspartic acid contributed relatively high in the protein and enzymatic peptides.The SDS-PAGE results showed that the sea buckthorn protein could be well hydrolyzed by alkaline protease,and the molecular weight of the enzymatic peptides was concentrated below 15 kDa.This study could provide a theoretical basis and scientific basis for the further development and utilization of sea buckthorn resources in food and health products.

关 键 词：沙棘籽粕 酶解多肽 乙醇脱氢酶 工艺优化 氨基酸组成 分子量 

分 类 号：TS209[轻工技术与工程—食品科学]