具有醒酒活性的玉米肽的制备、富集和鉴定  

Preparation,Enrichment and Identification of Corn Peptides with Facilitating Alcohol Metabolism Activity

作　　者：赵谋明[1] 马梅 苏国万[1] 郑淋[1] 刘洋[1] Zhao Mouming;Ma Mei;Su Guowan;Zheng Lin;Liu Yang(South China University of Technology,Guangzhou 510640)

机构地区：[1]华南理工大学,广州510640

出　　处：《中国食品学报》2020年第9期86-94,共9页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家重点研发计划(2017YFD0400200)。

摘　　要：为制备具有高效醒酒活性的玉米肽,采用木瓜蛋白酶和胰酶复合酶酶解玉米黄粉。以蛋白质回收率和乙醇脱氢酶(ADH)激活率为指标,确定最佳的酶解工艺。结果:木瓜蛋白酶和胰酶复合酶的最佳酶解条件是:料液比1∶10,总加酶量2%,木瓜蛋白酶和胰酶质量比1∶1,酶解时间8 h。在此条件下蛋白质回收率为69.89%,ADH激活率为65.72%。为进一步分离富集具有高醒酒活性的成分,先用不同体积分数浓度的乙醇富集玉米肽中的醒酒成分,其中以80%乙醇的醇沉组分的ADH激活率最高(75.08%);再用葡聚糖凝胶G-15(Sephadex G-15)对80%乙醇的醇沉组分进行第2步分离,共获得4个组分,其中G4组分的ADH激活率高达90.12%。这2种分离方法高效富集了具有醒酒活性的玉米肽;最后用超高压液相色谱串联质谱(UPLC-ESI-Q-TOF-MS/MS)分离鉴定具有高效醒酒活性的肽段结构,共得6条肽段。这些肽段都含丙氨酸,其中3条肽段含有亮氨酸。由于这2种氨基酸有利于醒酒,所以这6条肽段具有醒酒活性。本研究结果为深入研发玉米醒酒肽提供了理论基础。This experiment used two kinds enzymes(papain and pancreatin)to hydrolyze corn gluten meal for preparing corn anti-alcoholism peptides,the single factor experiment was aimed to explore the influence of different liquid-to-material,the quantity of enzyme,bi-enzyme papain/pancreatin and reaction time during the enzymolysis on the protein recovery ratio and the activation ratio of ADH of corn anti-alcoholism peptides.The optimum technology conditions were as follows:the liquid-to-material ratio was 10∶1,the quantity of enzyme was 2%,the bi-enzyme papain/pancreatin was 1∶1,reaction time was 8 h.Under such condition,the protein recovery ratio was 69.89%and the ADH activation ratio was 65.72%.Then corn anti-alcoholism peptides were separated and enriched by ethanol with various concentrations,and alcohol-sinking component with 80%ethanol had the highest ADH activation ratio(75.08%).Sephadex G-15 was used for the second step separation and enrichment of the ethanol precipitation component of 80%ethanol,the four peaks were eluted,the overall recovery ratio was 84.52%.the percentage points of 4 classes were as followed:24.36%,47.79%,3.25%,9.12%,respectively.From the anti-alcohol experiments of 4 classes,the ADH activation ratio of the G4 fraction was as high as 90.12%,so corn peptide with facilitating alcohol metabolism activity was enriched efficiently.Ultra-performance liquid chromatography tandem mass-spectrometry(UPLC-ESI-Q-TOF-MS/MS)was applied for structural identification of the G4 fraction separated by Sephadex G-15.Six peptides with length ranging from 7 to 9 amino acids were found by searching against the corn data from Uniport database,including GQAIILP,GGTALPA,QAKGILP,YQQPIIGGA,VAAGGPA,VAVGGVP,3 of them contained leucine,all of them contain alanine,These two amino acids played an important role in facilitating alcohol metabolism activity.In addition,Proline also had a positive effect on facilitating alcohol metabolism.In conclusion,it was suggested that the corn hydrolysates could be used in the potential

关 键 词：玉米肽 ADH激活率 酶解优化 醇沉 富集 质谱法 

分 类 号：TS213.4[轻工技术与工程—粮食、油脂及植物蛋白工程]