藏羊血多肽制备工艺优化及其抗氧化活性  

Optimization of the Preparation Process of Tibetan Sheep Blood Peptide and Its Antioxidant Activity

作　　者：刘鑫 赵成周 韩丽娟 罗唐君 桂林生 侯生珍[3] 陈秀梅 孙胜男 LIU Xin;ZHAO Chengzhou;HAN Lijuan;LUO Tangjun;GUI Linsheng;HOU Shengzhen;CHEN Xiumei;SUN Shengnan(Department of Pharmacy,Qinghai University,Xining 810001,Qinghai,China;College of Tibetan Medicine,Qinghai University,Xining 810016,Qinghai,China;College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,Qinghai,China)

机构地区：[1]青海大学药学系,青海西宁810001 [2]青海大学藏医学院,青海西宁810016 [3]青海大学农牧学院,青海西宁810016

出　　处：《食品研究与开发》2025年第5期114-121,211,共9页Food Research and Development

基　　金：青海省“帅才科学家负责制”项目(2022⁃NK⁃169⁃4);青海大学大学生创新训练计划项目(2024⁃QX⁃63)。

摘　　要：以藏羊血液为原料,通过优化酶解条件,制备藏羊血多肽并分析其抗氧化活性。使用风味蛋白酶、碱性蛋白酶、中性蛋白酶、胃蛋白酶、木瓜蛋白酶和胰蛋白酶分别对藏羊血液进行水解,并以水解度为指标筛选适宜蛋白酶,在此基础上,研究酶解时间、加酶量、酶解温度、料液比和pH值对水解度的影响;采用响应面分析法对酶解参数进行优化;使用中高压液相色谱对血肽进行初步分离纯化和制备,以DPPH自由基和ABTS+自由基清除率试验探究藏羊血液多肽的体外抗氧化活性。结果表明,藏羊血多肽最佳水解酶为风味蛋白酶,最佳酶解条件为水解时间5 h、酶解温度56℃、料液比1∶7(mg/mL)、加酶量1050.00 U/g。在此条件下,水解度可达(54.90±0.32)%;体外抗氧化试验结果显示,藏羊血多肽粗品在高浓度时对DPPH自由基和ABTS+自由基的清除率分别可达阳性对照V_(C)的77%和87%,IC_(50)分别为(17.97±1.07)mg/mL和(14.36±0.63)mg/mL,使用SpHerical C_(18)色谱柱将藏羊血肽分离为10个组分,分离后的组分Fr3显示出较好的抗氧化活性,超过阳性对照。Tibetan sheep blood was used as the raw material to prepare blood peptides by optimizing the enzymatic hydrolysis conditions,and their antioxidant activity was analyzed.Flavor protease,alkaline protease,neutral protease,pepsin,papain,and trypsin were used to hydrolyze Tibetan sheep blood,with the degree of hydrolysis as the indicator for selecting the appropriate protease.On this basis,the effects of hydrolysis time,enzyme concentration,hydrolysis temperature,material-to-liquid ratio,and pH on the hydrolysis degree were investigated.The enzymatic hydrolysis parameters were optimized using response surface methodology.Blood peptides were preliminarily separated,purified,and prepared using medium-and high-pressure liquid chromatography,and their in vitro antioxidant activity was explored using DPPH and ABTS+radical scavenging assays.The results showed that flavor protease was the optimal enzyme for hydrolyzing Tibetan sheep blood peptides,with the best hydrolysis conditions being a hydrolysis time of 5 h,a temperature of 56℃,a material-to-liquid ratio of 1∶7(mg/mL),and an enzyme concentration of 105.00 U/g.Under these conditions,the degree of hydrolysis reached(54.90±0.32)%.The in vitro antioxidant activity results showed that,at high concentrations,the crude Tibetan sheep blood peptides exhibited DPPH and ABTS+radical scavenging rates of 77%and 87%,respectively,compared to the positive control V_(C),with IC_(50) values of(17.97±1.07)mg/mL and(14.36±0.63)mg/mL.The Tibetan sheep blood peptides were separated into 10 fractions using a spherical C_(18) column,with the Fr3 showing better antioxidant activity than the positive control.

关 键 词：藏羊血 多肽 风味蛋白酶 水解度 分离纯化 抗氧化能力 

分 类 号：TS251.93[轻工技术与工程—农产品加工及贮藏工程]