Characterization of antifungal organic acids produced by Lactobacillus harbinensis K.V9.3.1Np immobilized in gellan–xanthan beads during batch fermentation

Antifungal activity of lactic acid bacteria (LAB) has been observed in many food fermentation processes. Organic acids produced during the growth of these microorganisms play a major role in antagonism towards molds. The present study characterizes antifungal compounds produced during growth of Lactobacillus harbinensis K.V9.3.1Np that was previously isolated from raw cow milk and found to possess strong antifungal activities. A pH-controlled batch fermentation in enriched milk protein medium with immobilized cells of this bacterium was carried out in the presence of disrupted cells of Debaryomyces hansenii UBOCC-A-211003 and Kluyveromyces lactis UBOCC-A-212021. After seven days of fermentation, cell-free culture supernatants with robust antifungal activities against D. hansenii, Penicillium expansum and Penicillium roqueforti were obtained. These cell-free supernatants contained high amounts of organic acids such as lactic and acetic acids as well as hexanoic acid (9 mM), which the latter revealed to be the most efficient antifungal compound. The antifungal activity of hexanoic acid was influenced by the environmental conditions including the nature of the matrix and the pH. Under the conditions tested, even if organic acids were key antifungal agents, they were only part of a complex mixture of various molecules that acted in a synergistic manner.