| Summary: | The aim of the investigation was the identification and initial study of lactic acid bacteria (LAB) isolated from spontaneously fermented (at 28 °C for 5 days) fruiting bodies of white button mushrooms (<i>Agaricus bisporus</i>). The isolated LAB were preliminarily characterized applying the MALDI-TOF Biotyper. Moreover, further phenotypical, genotypical characteristics as well as some functional and technological properties of the selected microorganisms (including the ability to produce exopolysaccharides, cell hydrophobicity, resistance to low pH, and bile salt) were also analyzed. Among autochthonous LAB (isolated from the tested mushroom raw material), <i>Leuconostoc mesenteroides</i> predominated in spontaneously fermented <i>A. bisporus</i>, while <i>Lactiplantibacillus paraplantarum, Lactiplantibacillus plantarum</i>, and <i>Lactococcus lactis</i> were less abundant. The highest dynamics of acidification of the mushroom material were exhibited by isolates EK55 and EK4 that, after 24 h of incubation, were able to decrease the pH of the raw material up to 5.06 ± 0.021 and 5.17 ± 0.015, respectively. Furthermore, the analysis of bacterial cell hydrophobicity indicated that the highest values of this parameter were noted for <i>L. plantarum</i> isolates EK12 (29.59 ± 0.7%), EK55 (28.75 ± 0.551%), and EK5 (27.33 ± 1.516%). It was revealed some of the analyzed LAB (especially isolates <i>L. plantarum</i> EK55 and <i>L. paraplantarum</i> EK4) exhibited functional and technological potential that might be used in the formulation of novel starter cultures.
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