Generation of antibacterial peptides from crude cheese whey using pepsin and rennet enzymes at various pH conditions

Abstract

Background: The current study aimed to prepare antimicrobial agents for food preservation from crude cheese whey. Crude cheese whey was digested with porcine pepsin, and calf and fungal rennets at various pHs. The digests were assessed for antibacterial activities against Escherichia coli and Bacillus subtilis.

Results: The calf rennet digest at pH 3.0 showed the highest antibacterial activity. Three antibacterial peptides that acted against B. subtilis lactoferrin f(20-30), and β-lactoglobulin f(14-22) and f(92-103) were identified in the calf rennet digest. Only lactoferrin f(20-30) also exerted bactericidal activity against E. coli. LC-MS/MS analysis revealed that the three peptides were generated by the porcine pepsin at pH 2.5, whereas the calf rennet generated them at a wider pH (pH 2.5-3.5). Fungal rennet generated only β-lactoglobulin f^14-22 at pH 3.5. The pepsin and calf rennet digests at pH 2.5 and 3.0, respectively, reduced the E. coli and B. subtilis populations by approximately 2 log at 6000 µg mL^-1 in milk at 4 °C.

Conclusion: The calf rennet and porcine pepsin digests of cheese whey, at a specific acidic pH, which can be prepared from food-grade materials, have the potential to be used as natural food preservatives due to the presence of the three antibacterial peptides. © 2018 Society of Chemical Industry.

Keywords: antibacterial peptide; cheese whey; lactoferrin; pepsin; rennet; β-lactoglobulin.