红曲糟源酶解蛋白肽的功能性评价

Functional Evaluation of Enzymatic Hydrolyzed Peptides from Hongqu Glutinous Rice Wine Grains Protein

【目的】研究不同蛋白酶对红曲糟酶解的效果,并对其酶解液进行功能性评价,为红曲糟源蛋白肽的研发制备提供理论支持。【方法】以提高蛋白含量后的红曲糟为原料,利用碱性蛋白酶、胰蛋白酶、动物蛋白酶、木瓜蛋白酶、菠萝蛋白酶、胃蛋白酶、复配酶制剂F106、酵母抽提酶等不同蛋白酶进行酶解,考察其酶解率、1,1-二苯基-2-三硝基苯肼(DPPH)和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除抗氧化能力、黄嘌呤氧化酶(Xanthine oxidase,XOD)和血管紧张素转化酶(Angiotensin converting enzyme,ACE)抑制活性等生物活性功能。【结果】蛋白酶解率最高的为动物蛋白酶和胰蛋白酶酶解处理,分别为(71.43±1.03)%和(70.20±0.32)%。DPPH自由基清除抗氧化能力最优的是胃蛋白酶、碱性蛋白酶和酵母抽提酶酶解处理,蛋白肽的半数效应浓度(Median effective concentration,EC50)分别为(2.78±0.34)mg·mL^(-1)、(3.02±0.03)mg·mL^(-1)、(3.24±0.65)mg·mL^(-1);ABTS自由基清除抗氧化能力、XOD抑制活性能力最优的均为胃蛋白酶和碱性蛋白酶酶解液,其蛋白肽的EC50值分别为(1.54±0.07)mg·mL^(-1)、(6.45±0.27)mg·mL^(-1)和(10.71±0.06)mg·mL^(-1)、(17.68±0.04)mg·mL^(-1),二者的XOD半数抑制指数分别为(1.28±0.01)、(1.78±0.03);ACE抑制活性最优的为碱性蛋白酶酶解液和木瓜蛋白酶酶解液,蛋白肽的EC50值均为(0.27±0.01)mg·mL^(-1),半数抑制指数分别为(118.40±3.53)、(98.35±1.95)。【结论】红曲糟源蛋白经不同蛋白酶酶解后的肽段具有不同的生物活性,其中以胃蛋白酶酶解的XOD抑制活性能力较强,碱性蛋白酶和木瓜蛋白酶酶解的ACE抑制活性较优。

【Objective】The study investigated the effects of various proteases on the enzymatic hydrolysis of protein from Hongqu glutinous rice wine grains,followed by a functional evaluation of the resulting enzymatic hydrolysate.The purpose was to provide theoretical support for the development and preparation of protein peptides derived from Hongqu glutinous rice wine grains protein.【Method】Purified Hongqu glutinous rice wine grains were utilized as the raw material,employing various proteases,including alcalase,trypsin,animal protease,papain,bromelain,pepsin,compound enzyme preparation F106,and yeast extract enzyme,for enzymatic hydrolysis.The enzymatic hydrolysis rate,1,1-diphenyl-2-picrylhydrazyl(DPPH),and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt(ABTS)free radical scavenging antioxidant ability,xanthine oxidase(XOD),and angiotensin-converting enzyme(ACE)inhibitory activity,along with other biological activity functions,were described.【Results】The peak proteolytic rates for animal protease and trypsin were(71.43±1.03)%and(70.20±0.32)%,respectively.Among the hydrolysates,those derived from pepsin,alcalase and yeast extract enzyme exhibited the best DPPH free radical scavenging antioxidant capacity.The median effective concentration(EC50)of peptides were(2.78±0.34)mg·mL^(-1),(3.02±0.03)mg·mL^(-1)and(3.24±0.65)mg·mL^(-1),respectively.The hydrolysates of pepsin and alcalase exhibited superior ABTS scavenging and XOD inhibition capabilities.The protein of peptides EC50 values were(1.54±0.07)mg·mL^(-1),(6.45±0.27)mg·mL^(-1),(10.71±0.06)mg·mL^(-1),and(17.68±0.04)mg·mL^(-1),respectively,while their half inhibitory indices of XOD were(1.28±0.01)and(1.78±0.03),respectively.Alcalase and papain hydrolysate showed themostoptimal ACE inhibitory activity.The EC50 of the peptides were(0.27±0.01)mg·mL^(-1)and their half inhibition indices were(118.40±2.53)and(98.35±1.95),respectively.【Conclusion】The peptides derived from the protein of Hongqu glutinous rice wine grains exhibit different biological activities after enzymatic hydrolysis by various proteases.Among them,the XOD inhibitory activity of pepsin enzymatic hydrolysis is stronger,and the ACE inhibitory activity of alcalase and papain enzymatic hydrolysis is better.