辣木籽凝乳酶在水牛乳干酪制备中应用及产血管紧张素转化酶抑制肽分析  

Application of Moringa oleifera Seed Rennet in the Preparation of Buffalo Cheese:Optimization of Processing Technology and Mining of Angiotensin Converting Enzyme Inhibitory Peptides

作　　者：钟玉旺 于梦怡 张丹 常莹 幸福兵 黄艾祥[1] 王雪峰[1] ZHONG Yuwang;YU Mengyi;ZHANG Dan;CHANG Ying;XING Fubing;HUANG Aixiang;WANG Xuefeng(College of Food Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Lijiang Vocational and Technical College,Lijiang 674100,China)

机构地区：[1]云南农业大学食品科学技术学院,云南昆明650201 [2]丽江职业技术学院,云南丽江674100

出　　处：《食品科学》2024年第18期87-98,共12页Food Science

基　　金：国家自然科学基金地区科学基金项目(32360562);云南省农业联合专项重点项目(202101BD070001-013);云南省科学技术协会2022—2024年度青年科技人才托举工程项目。

摘　　要：本实验以辣木籽凝乳酶和水牛乳为原料,以干酪感官评分和蛋白水解度为指标优化干酪加工工艺,解析其水溶性肽组成,进一步挖掘具有血管紧张素转化酶(angiotensin converting enzyme,ACE)抑制活性的功能性肽及其潜在分子作用机制。结果表明,干酪最佳工艺参数为凝乳pH 5.56、凝乳酶添加量0.14%、凝乳时间32 min、热烫温度80℃,此条件下干酪水解度为12.56%,所得干酪质地均匀有弹性、口感柔软细腻、色泽均匀有光泽且香味浓郁;基于液相色谱-串联质谱在干酪中检出1600种分子质量小于3 k Da的蛋白肽段,活性预测表明613条肽段具有较好的生物活性,主要来源于β-酪蛋白和α_(S1)-酪蛋白,其中具有ACE抑制活性的肽段占比达到37.29%;通过活性肽数据库比对结合生物信息学手段筛选出兼具多重活性的已知ACE抑制肽和新型ACE抑制肽各4条,分子对接分析进一步表明4条新型肽段均可以与ACE有效结合,其中2条肽段FGGL、FSPL分别与S2活性口袋结合紧密而显示较强的ACE抑制活性。本研究可为新型植物凝乳酶资源及特色水牛乳干酪的开发利用提供科学依据。In this study,buffalo cheese was prepared with rennet from Moringa oleifera seeds.The process was optimized based on sensory characteristics and hydrolysis degree.The composition of water-soluble peptides from the cheese was analyzed.Furthermore,bioactive peptides with angiotensin converting enzyme(ACE)inhibitory activity were identified and their potential molecular mechanisms of action were explored.The optimal conditions determined were as follows:pH 5.56,rennet dose 0.14%,incubation time 32 min,and stretching temperature 80℃.Under these conditions,the hydrolysis degree of cheese was 12.56%with a uniform and elastic texture,a soft and delicate taste,a uniform and glossy color,and a rich aroma.By liquid chromatography-tandem mass spectrometry(LC-MS/MS),1600 peptides with molecular mass less than 3 kDa were detected in the cheese,613 of which demonstrated good bioactivity,predominately derived fromβ-casein andα_(S1)-casein.Notably,37.29%of the peptides exhibited ACE inhibitory activity.By comparing with the bioactive peptide database and using bioinformatics,four peptides proven to have ACE inhibitory activity and four novel peptides with potential ACE inhibitory activity were identified.Molecular docking showed that the four novel peptides could effectively bind to ACE.Among them,FGGL and FSPL bound tightly to the S2 active pocket,thus exhibiting strong ACE inhibitory activity.This study provides a scientific basis for the development and utilization of new plant rennet resources and characteristic buffalo cheese.

关 键 词：辣木籽凝乳酶 水牛乳干酪 液相色谱-串联质谱 血管紧张素转化酶抑制肽 分子对接 

分 类 号：TS252.53[轻工技术与工程—农产品加工及贮藏工程]