复合酶提取石榴幼果总黄酮工艺优化及其抑制α-葡萄糖苷酶活性  

Complex Enzymatic Extraction of Total Flavonoids from Pomegranate Fruitlets and Their Anti-α-Glucosidase Activity

作　　者：王占一 廖成斌 公金艳 张卓 汤锐睿 戴博 WANG Zhan-yi;IAO Cheng-bin;GONG Jin-yan;ZHANG Zhuo;TANG Rui-rui;DAI Bo(College of Food Science and Pharmaceutical Engineering,Zaozhuang University,Zaozhuang 277160,China;Department of Pharmacy,Air Force General Hospital,PLA,Beijing 100142,China)

机构地区：[1]枣庄学院食品科学与制药工程学院,山东枣庄277160 [2]空军总医院药学部,北京100142

出　　处：《食品工业科技》2019年第9期178-183,188,共7页Science and Technology of Food Industry

基　　金：2015年度国家星火计划项目(2015GA740027);山东省自然科学基金(ZR2018LH020);山东省重点研发计划项目(2016GSF202010);山东省高等学校科技计划项目(J18KA305)

摘　　要：优化复合酶(纤维素酶-果胶酶-木瓜蛋白酶)提取石榴幼果总黄酮的工艺,并测定其抑制α-葡萄糖苷酶活性。以总黄酮得率为评价指标,在单因素实验基础上,D-最优混料设计优化复合酶配比,正交试验设计对料液比、介质pH、酶解温度、酶解时间进行优化,并以PNPG为底物测定总黄酮对α-葡萄糖苷酶的抑制活性。结果显示:复合酶最优配比为:纤维素酶44.2%、果胶酶31.6%、木瓜蛋白酶24.2%,最佳提取条件为:料液比1∶18 (g/mL)、介质pH5.0、酶解温度50℃、酶解时间4.0 h,总黄酮得率为3.38%,其浓度为1.5 mg/mL时,对α-葡萄糖苷酶抑制率达到63.9%,抑制作用的IC_(50)为1.059 mg/mL,在浓度0.15～1.5 mg/mL范围内,石榴幼果总黄酮浓度与其对α-葡萄糖苷酶抑制效果之间呈现一定的正相关关系,其抑制机理属于可逆性抑制和非竞争性抑制。该方法可为石榴幼果总黄酮的提取和应用提供一定的科学依据。To optimize the complex enzymatic (cellulose-pectinase-papain) extraction of total flavonoids from pomegranate fruitlets and to determine its anti-α-glucosidase activity.with the yield of total flavonoids as an evaluation index,D-optimal mixture design was applied to optimize the complex enzymeratio,and orthogonal experimental design was adopted in the optimization of solid/liquid ratio,medium pH,enzymatic hydrolysis temperature and enzymatic hydrolysis time based on the single factor test,then the anti-α-glucosidase activity was determined with PNPG as substrate. The results showed that optimal ratio of enzyme was determined to be 44.2% for cellulose,31.6% for pectinase and 24.2% for papain,to obtain a 3.38% of total flavonoids yield,the optimal conditions were determined to be 1∶18(g/mL)for solid/liquid ratio,5.0 for medium pH,50 ℃ for enzymatic hydrolysis temperature and 4.0 h for enzymatic hydrolysis time,the substrate inhibition rate reached 63.9% on α-glucosidase and the inhibitory effect of IC50 was 1.059 mg/mL at the centration was 1.5 mg/mL.There was a dose-response relationship between the total flavonoids and anti-a-glucosidaseactivity in the range of 0.15~1.5 mg/mL of concentration,the inhibition mechanism was reversible inhibition and non competitive inhibition. This method can provide ascientific basis for the extraction and application of total flavonoids from pomegranate fruitlets.

关 键 词：石榴幼果 总黄酮 复合酶法 D-最优混料设计 抑制α-葡萄糖苷酶活性 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]