Purification and Identification of Potentially Bioactive Peptides from Enzyme-Modified Cheese

¹ Dept. of Food Science and Agricultural Chemistry, McGill University, 21,111, Lakeshore Road, Ste-Anne-de-Bellevue, Quebec, Canada H9X 3V9
² Dept. of Food Science and Agricultural Chemistry, McGill University, 21,111, Lakeshore Road, Ste-Anne-de-Bellevue, Quebec, Canada H9X 3V9 and Food Research and Development Center, Agriculture and Agri-Food Canada, St-Hyacinthe, Quebec, Canada J2S 8E3

Antihypertensive peptides inhibiting angiotensin I-converting enzyme have been isolated from enzymatic hydrolysates of various food materials, but no information is available on the isolation of antihypertensive peptides from enzyme-modified cheese. In this study, several bioactive peptides, mainly potential antihypertensive peptides from enzyme-modified cheese prepared by commercial and Lactobacillus casei enzymes, were purified and identified. Enzymemodified cheese samples were prepared by combination of Neutrase® (1883.0 U/ml), L. casei enzymes (amino peptidase activity 86.4 leucine aminopeptidase U/g), and Debitrase^TM (22.0 leucine aminopeptidase U/g). The water-soluble fractions of the enzymemodified cheeses that were prepared by different enzymes were subjected to reverse-phase HPLC on a Delta Pak C₁₈ column. Each peak was purified on the same column using a binary gradient. One peak from the Neutrase® digest, five peaks from the Neutrase®- Debitrase^TM digest, and two peaks from the Neutrase®-Lactobacillus enzyme digest were purified and identified by API mass spectrometry. On the basis of their molecular masses, amino acid sequences of purified peptides were identified. ß-Casomorphin with a sequence like that of ß-casein (YPFPGPI f 60–66) was found after the Neutrase® digest. All of the peptides purified from the digests with combination of Neutrase® and Debitrase^TM or Neutrase® and L. casei enzymes contained active sites in their sequences. The presence of sites containing potential antihypertensive peptides suggests that the purified peptides may have antihypertensive properties. Thus, the enzyme-modified cheese process, mainly designed to produce flavor ingredients, may simultaneously produce bioactive peptides, which are considered to be of physiological importance.

Key Words: bioactive peptide • antihypertensive peptide • enzyme-modified cheese • Lactobacillus casei

Submitted on September 28, 1998
Accepted on April 5, 1999

This article has been cited by other articles:

				J. Meyer, U. Butikofer, B. Walther, D. Wechsler, and R. Sieber Hot topic: Changes in angiotensin-converting enzyme inhibition and concentrations of the tripeptides Val-Pro-Pro and Ile-Pro-Pro during ripening of different Swiss cheese varieties J Dairy Sci, March 1, 2009; 92(3): 826 - 836. [Abstract] [Full Text] [PDF]

				D. R. Henning, R. J. Baer, A. N. Hassan, and R. Dave Major advances in concentrated and dry milk products, cheese, and milk fat-based spreads. J Dairy Sci, April 1, 2006; 89(4): 1179 - 1188. [Abstract] [Full Text] [PDF]

				C. G. Rizzello, I. Losito, M. Gobbetti, T. Carbonara, M. D. De Bari, and P. G. Zambonin Antibacterial Activities of Peptides from the Water-Soluble Extracts of Italian Cheese Varieties J Dairy Sci, July 1, 2005; 88(7): 2348 - 2360. [Abstract] [Full Text] [PDF]

				G. S. Lv, G. C. Huo, and X. Y. Fu Expression of Milk-Derived Antihypertensive Peptide in Escherichia coli J Dairy Sci, June 1, 2003; 86(6): 1927 - 1931. [Abstract] [Full Text] [PDF]

				V. Vermeirssen, J. Van Camp, K. Decroos, L. Van Wijmelbeke, and W. Verstraete The Impact of Fermentation and In Vitro Digestion on the Formation of Angiotensin-I-Converting Enzyme Inhibitory Activity from Pea and Whey Protein J Dairy Sci, February 1, 2003; 86(2): 429 - 438. [Abstract] [Full Text] [PDF]