Effects of a lactoperoxidase-thiocyanate-hydrogen peroxide system on Salmonella enteritidis in animal or vegetable foods.

Lactoperoxidase (LPO) from skim milk was purified by ion-exchange chromatography. The purified protein was used to catalyze the oxidation of thiocyanate by H2O2 in an antibacterial system (LPO system). The LPO system was used to inactivate or inhibit Salmonella enteritidis in tomato juice, carrot juice, milk, liquid whole egg, and chicken skin extract under various conditions. The system was found to be more effective against the organism in vegetable juices than in animal products, at low pH than at neutral pH, and at higher temperatures than at lower temperatures. Acid-adapted S. enteritidis cells were more susceptible than nonadapted cells. The system reduced numbers of S. enteritidis in vegetable products by up to 5.4 log units and inhibited growth of the organism in animal-derived foods during 4 h incubation at 30 degrees C. Sodium chloride (>100 mM) and polyphosphate (0.01-0.5%) enhanced the antibacterial effects of the system in tomato juice and chicken skin extract, respectively. The findings indicate that the LPO system could probably be used to prevent the growth and survival of salmonellae in minimally processed fruit and vegetable products, but combination of the system with other preservatives or treatments would be needed to effectively inhibit growth and survival of salmonellae in animal products.

Preparation of antimicrobial reduced lysozyme compatible in food applications.

The structural and antimicrobial functions of lysozyme reduced with food-compatible reducing agents-cysteine (Cys) and glutathione (GSH)-were investigated. The disulfide bonds were partially reduced by thiol-disulfide exchange reactions under heat-induced denaturing conditions from 55 to 90 degrees C. The results showed that treatment of lysozyme with Cys and GSH resulted in the introduction of new half-cystine residues (2-3 residues/mol of protein). The released SH groups, in turn, rendered the lysozyme molecule more flexible, being accompanied by a dramatic increase in the surface hydrophobicity and exposure of tryptophan residues. As a consequence, the resulting reduced lysozymes were more capable of binding to lipopolysaccharides (LPS) and permeabilizing the bacterial outer membrane, as evidenced by the liposome leakage experiment, than were native or heated lysozyme. Both reduced lysozymes displayed significantly higher antimicrobial activity than native or heated lysozyme against Salmonella enteritidis (SE) in sodium phosphate buffer (10 mM, pH 7.2) at 30 degrees C for 1 h. Their minimal inhibitory concentrations (MICs) against the tested bacteria were about 150- and 25-fold lower than their respective MICs of native or heated lysozyme. The results suggest that partially reduced lysozyme could be used as a potential antimicrobial agent for prevention of SE attack.