0 [24]. These facts and findings suggest that some lactobacilli are able to tolerate a very alkaline environment in some occasions for MAPK Inhibitor Library their survival. Nishitani et al. [7] speculated that tannase production might allow L. plantarum strains to accumulate high intracellular levels of Mn2+, which is otherwise chelated by tannins, compensating for the absence of superoxide dismutase in L. plantarum and providing resistance to oxygen toxicity under aerobic conditions. If this is actually the case, the alkaline tolerant tannases may be beneficial for their survival under alkaline conditions. It was reported that tannase activities were affected by

several chemicals [10, 25]. The activities of recombinant TanLpl, TanLpa, and TanLpe were significantly affected by neither EDTA, Mn2+, Mg2+, Ca2+, PMSF, nor urea. Previously Curiel et al. [10] reported that the activity of TanLpl was greatly increased in the presence of Ca2+ and was significantly

inhibited in the presence of urea. Notably, they used E. coli as a host for the expression of a recombinant N-terminal His-tagged TanLpl while we used B. subtilis and C-terminal His-tagged recombinant. In order to clarify the inconsistency further characterization is required, but the effects of EDTA, Mn2+, and Mg2+ were in good agreement. TanLpl, TanLpa, and TanLpe were not affected in 1 mM EDTA, implying that the enzymes do not Roxadustat concentration depend on divalent metallic ions as co-factors. Only the exception was Fe2+, which was also shown to inhibit a tannase from Penicillium chrysogenum[26]. Previously [9]K m value of TanLpl for MG was found to be lower than that of commercially available tannase of A. orzae. In this study, K m and k cat values of TanLpl, TanLpa, and TanLpe were calculated not only for MG, but also for various catechin

galloyl esters. K m values of the enzymes on each these substrates were comparable; however, k cat/K m values of TanLpa for EGCg, ECg, Cg, and GCg were significantly higher than those of TanLpl and TanLpe. The results suggest that the small differences in the amino acid sequences of tannase can exert Mirabegron such a drastic effect. Interestingly, k cat/K m values for EGCg3″Me of TanLpl and TanLpa were much lower than those for other catechins (Table 2). EGCg3″Me is a derivative of EGCg, in which the methoxyl group is located at the galloyl group of EGCg. Ren et al. [19] showed that the hydrogen-bonding network which was observed between three hydroxyl groups of gallic acid and the side chains of Asp421, Lys343, and Glu357 of lactobacilli tannase is important in enzyme-substrate complex. Therefore, EGCg3″Me may not form a strong and stable complex with the enzymes. Tannins are widely distributed in the plant kingdom and bind readily with proteins or heavy metals to form insoluble complexes, thereby presumably acting as a defense mechanism in plants against microbial attacks [7, 27].