Purification, diagnosis of the siderophorus of the probiotic Lactobacillus plantarum BHC03 strain and investigation of their relationship with some probiotic properties

In this study, siderophore producing microorganisms were obtained from the laboratory culture collection and the strain determined to have the highest siderophore production efficiency of 92% identified by 16s rRNA sequencing as Lactobacillus plantarum and called the strain as L. plantarum BHC03. Then, siderophore production optimization of L. plantarum BHC03 strain was performed and the highest siderophore production, respectively; It was determined at 37 °C, pH 7, in the absence of FeCl 3 addition and using yeast extract as nitrogen source and maltose as carbon source. The initial siderophore production was increased by 5 % after the optimization of the L. plantarum BHC03 strain, which was 92 %, and reached 97 %. Large-scale production of siderophores was carried out using optimized conditions and concentrated for purification by lyophilization. The extracellularly produced siderophore was purified by size exclusion chromatography using sephadex LH-20 resin. TLC chromatography, FT- IR, HPLC and 1H-NMR analyzes were performed for the determination and characterization of the presence of siderophores in the collected fractions after purification. As a result of the analyzes, it was determined that the 3rd and 5th fractions in the collected fractions contained catecholate-type siderophores and the effect of the catecholate-type siderophore on the probiotic properties of L. plantarum BHC03 strain was investigated. As a result of in vitro tests; L. plantarum BHC03 strain was shown to have 0.5 % phenol tolerance, 0.3 % bile salt tolerance and 8 % NaCl tolerance. However, it showed resistance to life up to pH 2.5 and was found to have no hemolytic activities. In addition, they were found to have 92 % autoaggregation and 93 % hydrophobicity abilities. L. plantarum BHC03 strain was found to be resistant to streptomycin, gentamicin and levoflovacin. It has been determined that siderophores have a positive effect on cholesterol assimilation and increase the cholesterol assimilation ability of the bacteria. However, it has been determined that siderophores have positive effects on the hemolysis and bacteriocin activity of probiotics. As a result of in vitro tests, it has been proved that L. plantarum BHC03 strain has the desired conditions to be accepted as probiotic bacteria and that catecholate siderophores have positive effects on bacteriocin activity and hemolytic activity.