Production and transepithelial transportation of angiotensin-I-converting enzyme (ACE)-inhibitory peptides from whey protein hydrolyzed by immobilized Lactobacillus helveticus proteinase.
Lactobacillus helveticus LB 10 proteinases immobilized with sodium alginate were used to hydrolyze whey protein to produce angiotensin-I-converting enzyme (ACE)-inhibitory peptides. The generated hydrolysates were tested for ACE-inhibitory activity and for their ability to be transported across Caco-2 cell monolayers. Using a response surface method, we determined that a proteinase concentration of 7.55 mg/mL, sodium alginate concentration of 2.03 g/100 mL, and glutaraldehyde concentration of 0.39% were found to be the optimal immobilization conditions. Compared with free proteinase, the immobilized proteinase had significantly higher pH, thermal and storage stability, and reusability. Whey protein hydrolysates were fractionated by gel filtration chromatography and ACE-inhibitory peptide mixtures were transported across Caco-2 cell monolayers in a human intestinal-absorption model. The di- and tripeptides KA, EN, DIS, EVD, LF, AIV, and VFK (half-maximal inhibitory concentrations (mean ± standard deviation) of 1.24 ± 0.01, 1.43 ± 0.04, 1.59 ± 0.27, 1.32 ± 0.05, 1.60 ± 0.39, 2.66 ± 0.02, and 1.76 ± 0.09 mmol/L, respectively) were detected on the basolateral side of the Caco-2 cell monolayer using ultra-performance liquid chromatography-tandem mass spectrometry. These results highlight that ACE-inhibitory peptides are present on the basolateral side of the Caco-2 cell model after transportation of whey protein hydrolysate across the Caco-2 cell membrane.
['Guo|Yuxing|Y|', 'Jiang|Xiaoxiao|X|', 'Xiong|Binyi|B|', 'Zhang|Tao|T|', 'Zeng|Xiaoqun|X|', 'Wu|Zhen|Z|', 'Sun|Yangying|Y|', 'Pan|Daodong|D|']
D000806:Angiotensin-Converting Enzyme Inhibitors D000818:Animals D001692:Biological Transport D018938:Caco-2 Cells D002462:Cell Membrane D004800:Enzymes, Immobilized D006801:Humans D006868:Hydrolysis D052198:Lactobacillus helveticus D010447:Peptide Hydrolases D007703:Peptidyl-Dipeptidase A D059748:Proteolysis D000067816:Whey Proteins
angiotensin-I-converting enzyme (ACE)-inhibitory peptide Lactobacillus helveticus proteinase Caco-2 cell bioavailability