β-glucan formation is a selective advantage for beer-spoiling Levilactobacillus brevis TMW 1.2112 during planktonic growth

Some lactic acid bacteria (LAB) isolated from beer or wine produce capsular β-glucans from UDP-glucose via the membrane-anchored glycosyltransferase GTF-2. This phenomenon is feared in breweries, because the viscosity of the affected liquids drastically increases due to the β-glucan and concomitant pellicle formation of these LAB. Currently it is unknown if this type of polysaccharide formation provides any advantage for the producing LAB during the colonization of (ethanol-containing) liquids. We thus used the β-glucan producer Levilactobacillus (L.) brevis TMW 1.2112 and its β-glucan-deficient transposon mutant (Δ gtf-2), and compared their growth at different ethanol concentrations and their competitiveness during co-cultivation. No significant inhibition in growth and differences in acidification were observed for both strains up to ethanol concentrations of 8% (v/v). At 10 % ethanol, the β-glucan forming wildtype increased its cell number and produced more acid in comparison to the mutant strain, which settled at the bottom of the fermentation tubes at any tested condition. At higher ethanol concentrations (12–18 % v/v) both strains failed to grow, while a higher viability of the wildtype strain was observed. After co-cultivation of both strains for up to 72 h in liquid nutrient medium (without ethanol), significantly more ropy wildtype colonies were detected, if the wildtype had been initially applied in similar cell counts or in excess. By contrast, the number of smooth mutant colonies was solely significantly higher after 24 h of growth, if the mutant strain had been initially inoculated in excess. These results indicate that the β-glucan-mediated pellicle formation by L. brevis TMW 1.2112 is its dominant phenotype and a selective advantage during colonization of liquids.