Milk Proteins Are Predigested Within the Human Mammary Gland
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Previous work demonstrates that proteases present in human milk release hundreds of peptides derived from milk proteins. However, the question of whether human milk protein digestion begins within the mammary gland remains incompletely answered. The primary objective of this study was to determine whether proteolytic degradation of human milk proteins into peptides begins within the mammary gland. The secondary objectives were to determine which milk proteases participate in the proteolysis and to predict which released peptides have bioactivity. Lactating mothers (n = 4) expressed their milk directly into a mixture of antiproteases on ice followed by immediate freezing of the milk to limit post-expression protease activity. Samples were analyzed for their peptide profiles via mass spectrometry and database searching. Peptidomics-based protease prediction and bioactivity prediction were each performed with several different approaches. The findings demonstrate that human milk contains more than 1,100 unique peptides derived from milk protein hydrolysis within the mammary gland. These peptides derived from 42 milk proteins and included 306 potential bioactive peptides. Based on the peptidomics data, plasmin was predicted to be the milk protease most active in the hydrolysis of human milk proteins within the mammary gland. Milk proteases actively cleave milk proteins within the mammary gland, initiating the release of functional peptides. Thus, the directly breastfed infant receives partially pre-digested proteins and numerous bioactive peptides.
KeywordsPeptides Human Mother’s milk Casein Whey
The authors thank Melinda Spooner for assisting in collecting the human milk samples and Cora J. Dillard for editing the manuscript. SDN and DCD planned the study. SDN collected the samples. SDN and RB conducted the experiments and data analysis. SDN, RB and DCD prepared the manuscript. All authors read and approved the final manuscript. The authors acknowledge the Mass Spectrometry Center at Oregon State University, which is supported in part by the National Institute of Health grant S10OD020111 (CSM).
Supported by the K99/R00 Pathway to Independence Career Award, Eunice Kennedy Shriver Institute of Child Health & Development of the National Institutes of Health (R00HD079561) (DCD).
Compliance with Ethical Standards
This present study was approved and carried out in accordance with the guidelines from the Institutional Review Board at Oregon State University. All mothers gave their informed consent.
Conflict of Interest
The authors declare that they have no conflict of interest.
- 14.Rawlings ND, Barrett AJ, Bateman A. MEROPS: the database of proteolytic enzymes, their substrates and inhibitors. Nucleic Acids Res. 2011;40(D1):D343-D50.Google Scholar
- 15.Minervini F, Algaron F, Rizzello CG, Fox PF, Monnet V, Gobbetti M. Angiotensin I-converting-enzyme-inhibitory and antibacterial peptides from Lactobacillus helveticus PR4 proteinase-hydrolyzed caseins of milk from six species. Appl Environ Microbiol. 2003;69(9):5297 – 305.CrossRefPubMedPubMedCentralGoogle Scholar
- 16.Azuma N, Nagaune S-I, Ishino Y, Mori H, Kaminogawa S, Yamauchi K. DNA-synthesis stimulating peptides from human β-casein. Agric Biol Chem. 1989;53(10):2631–34.Google Scholar
- 26.Ihara S, Miyoshi E, Nakahara S, Sakiyama H, Ihara H, Akinaga A, et al. Addition of β1–6 GlcNAc branching to the oligosaccharide attached to Asn 772 in the serine protease domain of matriptase plays a pivotal role in its stability and resistance against trypsin. Glycobiology. 2004;14(2):139 – 46.CrossRefPubMedGoogle Scholar