A variant peptide of buffalo colostrum β-lactoglobulin inhibits angiotensin I-converting enzyme activity

β-lactoglobulin is a rich source of bioactive peptides. The LC-MS separated tryptic peptides of buffalo colostrum β-lactoglobulin (BLG-col) were computed based on MS–MS fragmentation for de novo sequencing. Among the selected peptides (P1–P8), a variant was detected with methionine at position 74 instead of glutamate. The sequences of two peptides were identical to hypocholesterolemic peptides whereas the remaining peptides were in accordance with buffalo milk β-lactoglobulin. Comparative sequence analysis of BLG-col to milk β-lactoglobulin was carried out using CLUSTALW2 and a molecular model for BLG-col was constructed (PMDB ID-PM0076812). The synthesized variant pentapeptide (IIAMK, m/z-576 Da) was found to inhibit angiotensin I-converting enzyme (ACE) with an IC50 of 498 ± 2 μM, which was rationalized through docking simulations using Molgrow virtual docker.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► LC-MS separation of tryptic peptides of colostrum β-lactoglobulin. ► De novo sequencing of peptides using CID/ETD based MS/MS fragmentation. ► Hypocholesterolemic peptides identified with single amino acid variation. ► ACE inhibitory activity of variant peptide was studied. ► Molecular model for BLG-col was constructed.