利用一种新型羧肽酶M32制备低苦味豌豆低聚肽  

Preparation of low bitterness pea oligopeptide by a new carboxypeptidase M32

作　　者：丁沈利 毛炳杰 陆信曜 诸葛斌 宗红 DING Shenli;MAO Bingjie;LU Xinyao;ZHUGE Bin;ZONG Hong(The Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,School of Biotechnology,Research Centre of Industrial Microbiology,Wuxi 214122,China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,工业微生物研究中心,江苏无锡214122

出　　处：《食品与发酵工业》2023年第1期154-159,共6页Food and Fermentation Industries

摘　　要：该文研究了利用一种新型羧肽酶M32制备低苦味豌豆低聚肽的方法并进行了功能特性分析。结果表明羧肽酶M32酶解的最适条件为60℃,pH 7,酶解时间2 h,底物质量浓度60 g/L;羧肽酶M32结合碱性蛋白酶制备的豌豆低聚肽(pea protein isolate hydrolysed by alkaline protease and carboxypeptidase M32,PPIACP)的溶解性和水解度分别为80.6%和15.4%,与碱性蛋白酶单酶制备的豌豆低聚肽(pea protein isolate hydrolysed by alkaline protease,PPIA)相比,分别提高了37.4%和33.4%;PPIACP中分子质量在0.2~1 kDa的短肽与PPIA相比显著增加(P<0.05),增加了36.2%;鲜味氨基酸提高了3.8%,苦味氨基酸His、Arg、Val、Leu含量显著增加了17.4%(P<0.05),表明羧肽酶M32可将苦味肽C端的疏水性氨基酸(多为苦味氨基酸)释放出来,使其呈游离状态,降低苦味;电子舌结果显示,苦味值由5.09变为3.93,降低了22.8%;鲜味由18.9变为21.7,提高了20.9%;DPPH自由基清除率、•OH清除率分别为98%和90%。综上所述,结合羧肽酶M32的双酶法可以显著降低豌豆低聚肽的苦味并提高其功能特性,该羧肽酶在植物蛋白生物活性肽制备中具有良好的应用前景。A new carboxypeptidase M32 was used for the preparation and functional characteristics of low bitterness pea oligopeptide.The results showed that the optimum condition for enzymatic hydrolysis by carboxypeptidase M32 were 60℃,pH 7,enzymatic hydrolysis time 2 h and substrate mass concentration 60 g/L.The solubility and hydrolysis degree of pea protein isolate hydrolysed by alkaline protease and carboxypeptidase M32(PPIACP)were 80.6%and 15.4%,respectively,which were 37.4%and 33.4%higher than those of pea protein isolate hydrolysed by alkaline protease(PPIA).The oligopeptide between 0.2-1 kDa in PPIACP were significantly increased(P<0.05)by 36.2%compared to PPIA.The content of bitter amino acids,His,Arg,Val,and Leu,were significantly increased(P<0.05)by 17.4%,and that of umami amino acids were increased by 3.8%.The electronic tongue analysis showed that the bitterness value changed from 5.09 to 3.93,a decrease of 22.8%.The umami changed from 18.9 to 21.7,an increase of 20.9%.The DPPH radical scavenging activities and the hydroxyl radical scavenging activities were 98%and 90%,respectively.In conclusion,the dual enzymatic method combined with carboxypeptidase M32 can significantly reduce the bitterness of pea oligopeptide and improve their functional characteristics.This carboxypeptidase has a good application in the preparation of plant protein bioactive peptides.

关 键 词：羧肽酶M32 豌豆分离蛋白 抗氧化 脱苦 风味 

分 类 号：TS201.2[轻工技术与工程—食品科学]