Isolation and Structural Analysis of Antihypertensive Peptides That Exist Naturally in Gouda Cheese

Abstract

Seven kinds of ripened cheeses (8-mo-aged and 24-mo-aged Gouda, Emmental, Blue, Camembert, Edam, and Havarti) were homogenized with distilled water, and water-soluble peptides were prepared by C-18 hydrophobic chromatography. The inhibitory activity to angiotensin I-converting enzyme and decrease in the systolic blood pressure in spontaneously hypertensive rats were measured before and after oral administration of each peptide sample. The strongest depressive effect in the systolic blood pressure (-24.7 mm Hg) and intensive inhibitory activity to angiotensin I-converting enzyme (75.7%) were detected in the peptides from 8-mo-aged Gouda cheese. Four peptides were isolated by HPLC with reverse-phase and gel filtration modes. Their chemical structures and origins, clarified by combination analyses of protein sequencing, amino acid composition, and mass spectrometry, were as follows: peptide A, Arg-Pro-Lys-His-Pro-Ile-Lys-His-Gln [alpha(s1)-casein (CN), B-8P; f 1-9]; peptide B, Arg-Pro-Lys-His-Pro-Ile-Lys-His-Gln-Gly-Leu-Pro-Gln (alpha(s1)-CN, B-8P; f 1-13); peptide F, Tyr-Pro-Phe-Pro-Gly-Pro-Ile-Pro-Asn (beta-CN, A2-5P; f 60-68); and peptide G, Met-Pro-Phe-Pro-Lys-Tyr-Pro-Val-Gln-Pro-Phe (beta-CN, A2-5P; f 109-119). Peptides A and F, which were chemically synthesized, showed potent angiotensin I-converting enzyme inhibitory activity with little antihypertensive effects.

Biotechnological richness of the northeastern semi-arid region: antioxidant activity of casein hydrolysates from Moxotó goat milk (Capra hircus Linnaeus, 1758) obtained by papain action

“…Saito et al (2000) and Gómez-Ruiz, Ramos and Recio (2004) obtained casein peptides with similar molar mass molecular weight; nevertheless, they registered no antioxidant effect, as described in our study. They noted, however, antihypertensive properties.…”

“…Another peptides detected in our study were 887, 970 and 978 Da, earlier obtained by Saito et al (2000), Gómez-Ruiz, Ramos and Recio (2004) and Gómez-Ruiz et al (2008) from bovine casein hydrolysates. These peptides demonstrated antihypertensive and antioxidant characteristics.…”

Formation and Degradation of Beta-casomorphins in Dairy Processing

“…The levels of BCM7 and other related BCMs reported in some cheeses are shown in Table 2. In addition to BCM7, BCM5 was found in Brie, Rokpol, Edamski, Gouda, and Kasztelan (Sienkiewicz-Sz»apka et al, 2009), BCM9 (Saito et al, 2000;Toelstede and Hofmann, 2008) and BCM10 in Gouda (Toelstede and Hofmann, 2008) and BCM 11 in Caprino del Piemonte, an Italian goat cheese (Rizzello et al, 2005). In contrast, Muehlenkamp and Warthesen (1996) reported that BCM7 were not detected in Brie and Cheddar.…”

Bioactive peptides generated in an enzyme membrane reactor using Bacillus lentus alkaline peptidase

“…The IC 50 of V-b-casomorphin-9 against ACE was determined to be 325 lM. In the case of V-b-casomorphin-9, Saito et al [30] previously published an IC 50 value of 14.8 lM for b-casomorphin-9 (YPFPGPIPN), which exhibits a structural resemblance to V-b-casomorphin-9 (VYPFPGPIPN). Therefore, we conclude that additional Val at the N-terminal end of V-b-casomorphin-9 increased the IC 50 value almost 22-fold from 14.8 to 325 lM.…”