Lactic acid bacteria bioprotection applied to the malting process. Part I: Strain characterization and identification of antifungal compounds

Lactic acid bacteria (LAB) are of interest to cereal and beverage industry due to their contribution to the microbial safety and quality of fermented food/beverages. The main objectives of this study were to optimize the production of an antifungal cell-free-supernatant (cfs), based on lactic acid bacteria, using a wort-base substrate; and the subsequent identification of its active acid-base antifungal compounds. Two antifungal strains (Lactobacillus amylovorus DSM19280 and Lactobacillus reuteri R29) and a negative control strain (L. amylovorus DSM20552) were used. Barley based malt extract (wort) was produced and used as a fermentation substrate. Worts fermented with LAB were characterized in detail (total cell counts, pH, and total titratable acids (TTA) over time (0-120 h). A microtiter plate and an overlay agar based assay were used to evaluate the antifungal activity of the LAB cfs. Sugars, organic acids of the fermented and non-fermented wort (control) were analysed by HPLC. LAB antifungal metabolites were quantified using a QuEChERS method and an HPLC-UV/PDA system. Results show that wort produced from barley malt is a suitable substrate for LAB and dependent on the species and the fermentation time. The type of antifungal compounds can vary significantly. L. amylovorus DSM19280 cfs inhibited Fusarium culmorum spores at levels of 104 spores.mL−1, for 7 days, whereas L. reuteri R29 cfs inhibited up to 105 spores.mL−1 for the same time. Thirteen acid-base antifungal compounds were identified in L. reuteri R29 cfs and twelve in L. amylovorus DSM19280. Among them, phenyllactic, OH-phenylactic and benzoic acids were present at a significant concentration. This study demonstrates that the LAB wort-base cfs exhibits potent antifungal activity and is the ideal substrate for applications in a wide range of cereal products.