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液相色谱-串联质谱法分析牛κ-酪蛋白糖巨肽

Analysis of Bovine Kappa-Casein Glycomacropeptide by Liquid Chromatography-Tandem Mass Spectrometry.

作者信息

Qu Yunyao, Kim Bum-Jin, Koh Jeewon, Dallas David C

机构信息

Department of Food Science & Technology, Oregon State University, Corvallis, OR 97331, USA.

Nutrition Program, School of Biological and Population Health Sciences, College of Public Health and Human Sciences, Oregon State University, Corvallis, OR 97331, USA.

出版信息

Foods. 2021 Aug 28;10(9):2028. doi: 10.3390/foods10092028.

DOI:10.3390/foods10092028
PMID:34574138
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8469635/
Abstract

Caseinomacropeptide (CMP) is released from bovine kappa-casein after rennet treatment and is one of the major peptides in whey protein isolate. CMP has in vitro anti-inflammatory and antibacterial activities. CMP has two major amino acid sequences with different modifications, including glycosylation, phosphorylation and oxidation. However, no previous work has provided a comprehensive profile of intact CMP. Full characterization of CMP composition and structure is essential to understand the bioactivity of CMP. In this study, we developed a top-down glycopeptidomics-based analytical method to profile CMP and CMP-derived peptides using Orbitrap mass spectrometry combined with nano-liquid chromatography with electron-transfer/higher-energy collision dissociation. The liquid chromatography-tandem mass spectrometry (LC-MS/MS) spectra of CMPs were annotated to confirm peptide sequence, glycan composition and other post-translational modifications using automatic data processing. Fifty-one intact CMPs and 159 CMP-derived peptides were identified in four samples (one CMP standard, two commercial CMP products and one whey protein isolate). Overall, this novel approach provides comprehensive characterization of CMP and CMP-derived peptides and glycopeptides, and it can be applied in future studies of product quality, digestive survival and bioactivity.

摘要

酪蛋白巨肽（CMP）是在凝乳酶处理后从牛κ-酪蛋白中释放出来的，是乳清蛋白分离物中的主要肽类之一。CMP具有体外抗炎和抗菌活性。CMP有两个经过不同修饰的主要氨基酸序列，包括糖基化、磷酸化和氧化。然而，以前的工作都没有提供完整CMP的全面概况。对CMP的组成和结构进行全面表征对于理解CMP的生物活性至关重要。在本研究中，我们开发了一种基于自上而下的糖肽组学分析方法，使用Orbitrap质谱仪结合纳升液相色谱与电子转移/高能碰撞解离来分析CMP和CMP衍生的肽。利用自动数据处理对CMP的液相色谱-串联质谱（LC-MS/MS）谱进行注释，以确认肽序列、聚糖组成和其他翻译后修饰。在四个样品（一个CMP标准品、两个商业CMP产品和一个乳清蛋白分离物）中鉴定出了51种完整的CMP和159种CMP衍生的肽。总体而言，这种新方法提供了对CMP以及CMP衍生的肽和糖肽的全面表征，并且可应用于未来产品质量、消化存活率和生物活性的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/219deb055530/foods-10-02028-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/4030cbdc8922/foods-10-02028-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/ce5e598e08bc/foods-10-02028-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/2573b854e7d3/foods-10-02028-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/219deb055530/foods-10-02028-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/4030cbdc8922/foods-10-02028-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/ce5e598e08bc/foods-10-02028-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/2573b854e7d3/foods-10-02028-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5cfe/8469635/219deb055530/foods-10-02028-g004.jpg

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