Experiments were initially performed in shake flasks to identify the most suitable carbon source for maximizing the yield of biomass and lactic acid, and sucrose and glucose were chosen for further small scale batch experiments. As shown in Table 2 the growth rate of GDC-0449 ic50 L. crispatus L1 was not affected by the two different carbon sources; a slightly lower Yp/s was obtained with glucose, nevertheless, the latter is often preferred for industrial processes and therefore it was selected for the following fermentation experiments. In order to increase the production of biomass and related product a high cell density fermentation process exploiting a microfiltration strategy was developed to

keep the concentration of lactic acid below the toxic threshold for L .crispatus L1 (estimated to be 45 g · l−1, Figure 3). The feeding strategy avoided the waste of carbon source and determined a 7-fold and a 4-fold increase of the final titer of biomass and lactic acid, respectively, compared to previous batch experiments (Table 3). Based on earlier studies on L. bulgaricus[34] a higher improvement of the final biomass concentration was expected. Probably the VX-689 order adhesion of cells to membrane capillaries lowered transmembrane fluxes thus reducing the medium exchange rate. However, the concentration of biomass reached was very high compared to that obtained by cultivating other

lactobacilli; moreover, biomass resulted extremely viable (94%) at the end of the experiments (data not shown), valuable result for the foreseen application in medical devices/ food supplements. Adhesion seems C59 wnt purchase to be one of the key factors determining the colonization of the digestive ecosystem. Consistently the surface characteristics of lactobacilli are expected to contribute in several ways to their interactions with the host gastrointestinal tract and the gut microbiota, affecting their survival, adherence to the host tissue and interactions with themselves and with other bacteria. Since EPS can have important influences on these processes and on the colonization of the host [35, 36] we

also have investigated the chemical nature of the EPS produced by L. crispatus L1. This structure resulted to be a very intricate comb-like mannan polysaccharide that Casein kinase 1 has been already isolated and identified as capsule/EPS/protein bound-EPS in a number of microorganisms, among these in the yeast C. albicans[37]. We therefore hypothesised that the similarity of structure between the EPS of L. crispatus L1 and the carbohydrate part of mannoproteins and protein bound-polysaccharides excreted by C. albicans could be in part responsible for contrasting C. albicans infections. For this reason the ability of L. crispatus L1 live cells or of the purified EPS to hinder growth of C. albicans was analysed by performing adhesion assays with vaginal cells.