温福丽1,2，曾 超3，叶青青1,2，李 锋4，王松涛1,2*，沈才洪1,2
（1.泸州品创科技有限公司，四川泸州 646000；2.国家固态酿造工程技术研究中心，四川泸州 646000；
3.重庆大学生物工程学院，重庆 400030；4.郑州轻工业大学食品与生物工程学院，河南郑州 450001）
摘要：通过比较5种酶的水解情况筛选出一种最佳酶，并利用该酶水解得到的牡蛎酶解液经高效液相色谱-串联质谱分析鉴定牡蛎中的肽段及来源蛋白，利用在线数据库及构效关系进行活性肽筛选。结果表明，复合蛋白酶水解效果最佳。通过高通量分析牡蛎小肽中共得到427个小分子肽段，主要来源于肌动蛋白。经在线数据库活性筛选得到评分较高的8个降高血压活性肽：YPLRP、LPWRP、KYPLPR、FLPPR、VPFPR、FRCPE、FRCEP和HQGPLGPR，有望开发成ACE抑制肽。通过高通量筛选及在线数据库结合分析，实现复杂基质样品中大量肽段的高效活性筛选，为进一步研究牡蛎肽的功能和特点提供了理论依据。
关键词：牡蛎；复合蛋白酶；高效液相质谱联用；ACE抑制肽
中图分类号：TS254.4 文献标识码：A 文章编号：1674-506X（2023）03-0016-0008
Preparation of Crassostrea Hydrolysate and Identification of
Oligopeptides by High Throughput of HPLC-MS/MS
WEN Fuli1,2, ZENG Chao3, YE Qingqing1,2, LI Feng4, WANG Songtao1,2*, SHEN Caihong1,2
(1.Luzhou Pinchuang Technology Co., Ltd., Luzhou Sichuan 646000,China；
2.National Engineering Technology Research Center of Solid-state Brewing,Luzhou Sichuan 646000,China;
3.Bioengineering Institute of Chongqing University,Chongqing 400030,China;
4.College of Food and Bioengineering of Zhengzhou University of Light Industry,Zhengzhou Henan 450001,China)
Abstract：By comparing the hydrolysis conditions of the five enzymes, an optimal enzyme was selected. The enzymolysis solution obtained by hydrolysis was analyzed by High Performance Liquid Chromatography-Mass/Mass spectrometry (HPLC) to identify the peptides and source proteins in oyster. Activity prediction analysis was conducted using online database and structure-activity relationship. The hydrolysis effect of complex protease is good and suitable for industrial scale-up. A total of 427 small molecule peptides were identified from oyster,which mainly from actin protein tissues. The high score antihypertensive peptides YPLRP, LPWRP, KYPLPR, FLPPR, VPFPR, FRCPE, FRCEP and HQGPLGPR were obtained by online database activity screening,which were expected to be developed into good antihypertensive peptides in oyster. Through high-throughput screening and online database analysis, a large number of peptides in complex matrix samples could be screened with high efficiency, which provides a theoretical basis for further study of the function and characteristics of oyster peptides.
Keyword：Crassostrea gigas; complex protease; HPLC-MS/MS; ACE inhibitory peptide
doi：10.3969/j.issn.1674-506X.2023.03-003
收稿日期：2022-08-02
基金项目：四川省科技项目（2021ZYD0102）
作者简介：温福丽（1988-），女，硕士，工程师。研究方向：食品原料研究与开发。
*通信作者：王松涛（1977-），男，正高级工程师。研究方向：酿酒微生物与酿酒工艺研究，食品原料开发。
引用格式:温福丽,曾超,叶青青,等.牡蛎酶法制备及基于高通量分析鉴定小肽成分[J].食品与发酵科技,2023,59(3):16-22,29.
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