Generation of antimicrobial peptides Leg1 and Leg2 from chickpea storage protein, active against food spoilage bacteria and foodborne pathogens

Abstract

Contamination with bacteria leads to food waste and foodborne diseases with severe consequences for the environment and human health. Aiming to reduce food spoilage and infection, the present study developed novel highly active food-grade antimicrobial peptides affecting a wide range of bacteria. After extraction from chickpea, the storage protein legumin was hydrolyzed by the digestive protease chymotrypsin. Subsequent analysis by ultrahigh-performance micro-liquid chromatography-triple quadrupole time-of-flight tandem mass spectrometry determined the resulting peptide profiles. Virtual screening identified 21 potential antimicrobial peptides in the hydrolysates. Among those, the peptides Leg1 (RIKTVTSFDLPALRFLKL) and Leg2 (RIKTVTSFDLPALRWLKL) exhibited antimicrobial activity against 16 different bacteria, including pathogens, spoilage-causing bacteria and two antibiotic-resistant strains. Leg1/Leg2 showed minimum inhibitory concentrations (MIC) down to 15.6 µmol/L and were thus 10-1,000-fold more active compared to conventional food preservatives. Moreover, Leg1 and Leg2 showed bactericidal activity in contrast to the bacteriostatic activity of conventional preservatives.

Keywords: Antimicrobial peptides; Chickpea proteins; Chymotryptic hydrolysis; Cicer arietinum L.; Preservative; Virtual screening.