Bacteriocin bavaricin-A

Three hundred and thirty-five lactic acid bacteria were isolated from sour doughs and screened for antagonistic activity. Of these 145 showed activity against one or several of the indicator strains used in the screening. The antimicrobial activity of 18 isolates were due to a proteinaceous compound. These 18 isolates belonged to three different Lactobacillus species: Lactobacillus bavaricus, Lactobacillus curvatus and Lactobacillus plantarum. The spectrum of antimicrobial activity for the three species suggested that the inhibitory components were different. The inhibitory compound from Lact. bavaricus MI401 was chosen for further study. The proteinaceous nature, antimicrobial activity against closely-related species, heat resistance and sensitivity to alkaline treatment strongly indicated that this substance was a bacteriocin, which we designated bavaricin A. The bacteriocin was purified to homogeneity by ammonium sulphate precipitation, ion exchange, hydrophobic interaction and reverse-phase chromatography. The purification resulted in 193,000-fold increase in specific activity. SDS-PAGE of bavaricin A showed a molecular weight of 3500-4000 Da. By amino acid sequencing 41 amino acids were determined. Bavaricin A had a bactericidal mode of action and inhibited nine out of 10 Listeria monocytogenes. Lactobacillus bavaricus MI401 produced bavaricin A at temperatures from 4 degrees C to 30 degrees C. The production of active bavaracin A was inhibited at increasing sodium chloride concentration. In the presence of 3% sodium chloride at 4 degrees C no active bavaricin A could be detected. Nitrite (100 ppm) did not affect the production of active bavaricin A.

Several sourdough lactic acid bacteria (LAB) produce inhibitory substances other than organic acids. Bacteriocins (bavaricin A, and plantaricin ST31), a bacteriocin-like inhibitory substance (BLIS C57), and a new antibiotic (reutericyclin) have been discovered. Maximum antimicrobial production was found in the pH range 4.0-6.0. Temperature optima vary strongly. The substances are resistant to heat and acidity, and inactivated by proteolytic enzymes, except for reutericyclin. Bavaricin A and plantaricin ST31 have been purified to homogeneity. Bavaricin A is classified as a class IIa bacteriocin. Reutericyclin is a new tetramic acid. The mode of action of bavaricin A, BLIS C57, and reutericyclin is bactericidal. Some of these substances are active towards some Bacilli, Staphylococci and Listeria strains. Up to now, only the application potential of purified bavaricin A has been examined. More research should be done to study the production, the activity, and the stability of these inhibitory substances in food systems as these often differ from the broths mostly used in this kind of studies. Furthermore, an extensive screening of the sourdough microflora must be performed, in particular towards Bacilli and fungi. This could lead to the discovery of additional inhibitory substances, although it seems that the frequency of isolating bacteriocin-producing sourdough LAB is low. However, potent antimicrobials towards Bacilli as well as antifungal substances will have to be found using rational screening strategies and novel purification and analytical techniques.

In brined shrimp (ca. 3% NaCl), the effects of three different lactic acid bacteria bacteriocins (crude [6.54 x 10(sup10) U of bacteriocin activity {BU}/g] and purified [8.13 x 10(sup23) BU/g] nisin Z, carnocin UI49 [2.32 x 10(sup4) BU/g], and crude bavaricin A [2.78 BU/g]) on bacterial growth and shelf life were compared with those of a benzoate-sorbate solution (0.1% each [wt/wt]) and a control with no preservatives. The shelf life of shrimp subjected to the control treatment was found to be 10 days. Carnocin UI49 did not extend the shelf life, while crude bavaricin A (a cell-free supernatant of Lactobacillus bavaricus MI 401) resulted in a shelf life of 16 days, as opposed to 31 days with nisin Z for both its crude and purified forms. The benzoate-sorbate solution preserved the brined shrimp for the whole storage period (59 days). In the control, carnocin UI49, and crude bavaricin A treatments, a gram-positive flora dominated towards the end of the storage period while in the nisin Z treatment a gram-negative flora was more pronounced.