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In situ generation of milk protein-derived peptides in drying-off cows

Published online by Cambridge University Press:  08 September 2010

Chin H Ho
Department of Animal Science, National Chung Hsing University, Taichung 402, TaiwanROC
Chai J Chang*
Department of Animal Science, National Chung Hsing University, Taichung 402, TaiwanROC
Wen B Liu
Department of Animal Science, National Chung Hsing University, Taichung 402, TaiwanROC
Huo C Peh
Department of Animal Science, National Chung Hsing University, Taichung 402, TaiwanROC
Shuen E Chen
Department of Animal Science, National Chung Hsing University, Taichung 402, TaiwanROC
Hui Y Chen
Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan, ROC
Tsung H Ho
Department of Animal Science, National Chung Hsing University, Taichung 402, TaiwanROC
Ming T Chen
Department of Bioindustry Technology, Da Yeh University, Chung Hwa 555, Taiwan, ROC
Hajime Nagahata
Department of Animal Health, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501, Japan
*For correspondence; e-mail:


Our previous studies demonstrated prompt elevation of proteinase activity in mammary secretion of drying-off cows and goats. The current study examined the progressive changes in composition of cow mammary secretion following drying-off and, in parallel, characterized the mode of peptide neogenesis using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography-electrospray-ionization (LC-ESI) MS/MS. The results show that the percentage of casein of total milk protein at time of drying-off was 76%, which dropped to 41%, 24%, and 16%, respectively, 1, 2, and 3 weeks after drying-off. Levels of β-lactoglobulin and α-lactoalbumin in mammary secretions of drying-off cows decreased prominently while levels of lactoferrin, BSA, and casein derived-proteins increased concomitantly compared with regular milk. A fractionation procedure was applied to remove molecules larger than 10 kDa before MALDI-TOF MS and LC-ESI MS/MS and the results show that the MALDI-TOF MS peptide profile of mammary secretion ranging from m/z 600 to 4000 was apparently modified after drying-off for 1 week, whereas species 1590 m/z and 2460 m/z were most obviously enriched compared with regular milk. LC-ESI MS/MS results were used to map peptide sequence with Mascot search server and under no post translational modification to reduce database size and 202 novel β-casein-derived peptides were successfully identified in mammary secretion after drying-off for 1 week in contrast to regular milk. Accordingly at least 48 additional cleavage positions were assigned on β-casein for mammary secretion. Among the 202 novel peptides, 5 are homologous with confirmed opioid agonists, angiotensin 1-converting enzyme inhibitors, or immuno-modulators. In conclusion, peptides are released in situ from milk proteins within short intervals following drying-off in cows. They might play roles in the transition of mammary glands from lactating to non-lactating. With specified post-translational modifications and focused functional screening, novel peptides are yet to be discovered in dry cow mammary secretion.

Research Article
Copyright © Proprietors of Journal of Dairy Research 2010

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