A fibrinolytic enzyme of the mushroom, Schizophyllum commune was purified with chromatographic methods, including a DEAE-Sephadex A-50 ion-exchange column and gel filtrations with Sephadex G-75 and Sephadex G-50 columns. The analysis of fibrin-zymography and SDS-PAGE showed that the enzyme was a monomeric subunit that was estimated to be approximately 17 kDa in size. The fibrinolytic activity of the enzyme in plasminogen-rich and plasminogen-free fibrin plates was 1.25 and 0.44 U/ml, respectively. The N-terminal amino acid sequence of the purified enzyme was identified as HYNIXNSWSSFID, which was highly distinguished from known fibrinolytic enzymes. The relative activity of the purified enzyme with an addition of 5 mM EDTA, Phosphoramidon, and Bestatin was about 76, 64, and 52%, respectively, indicating that it is a metalloprotease. The optimum temperature for the purified enzyme was approximately 45°C, and over 87% of the enzymatic activity was maintained as a stable state in a pH range from 4.0 to 6.0. Therefore, our results suggest that the potential thrombolytic agent from S. commune is a unique type of fibrinolytic enzyme.
Aim: The objective of this study was to investigate toluene‐induced accumulation mechanism of trehalose in a toluene‐tolerant bacterium Pseudomonas sp. BCNU 106.
Methods and Results: The accumulation of trehalose by a toluene‐tolerant bacterium Pseudomonas sp. BCNU 106 was examined at various cultivation time by measuring the total intracellular trehalose content, trehalase activity and mRNA levels of the trehalose‐biosynthetic genes. The pattern of trehalose accumulation corresponded to the mRNA expression pattern of the trehalose‐biosynthetic genes with the maximum level at 12 h or 4 h of cultivation with 10% (v/v) toluene, respectively. The trehalose‐biosynthetic genes were also cloned and sequenced. Furthermore, the effects of toluene addition on the intracellular osmotic pressure and pH were investigated. It was shown that homeostasis was maintained in the bacterial cells.
Conclusions: In a toluene‐tolerant bacterium Pseudomonas sp. BCNU 106, a significant amount of trehalose was accumulated through the toluene‐induced expression of the trehalose‐biosynthetic genes after the exposure to toluene.
Significance and Impact of the Study: The accumulation of the high level of intracellular trehalose was preceded by the expression of otsA/B genes in toluene‐tolerant bacteria, contributing to the elucidation of the tolerance mechanism.
Lactic acid bacteria are generally recognized as beneficial probiotic organisms. Recent studies revealed that the potential of probiotic strains was essentially dependent on the bacterial-binding and adhesion capabilities to gut epithelial cells and the hydrophobicity of the cell surface. In this study, we screened some indigenous lactic acid bacteria from Kimchi and selected one lactic acid bacterium as a potential probiotic based on its cell surface hydrophobicity. Analysis of the 16S rRNA gene sequences of probiotic isolates indicated that the selected isolate (BCNU 9070 strain) was a member of Pediococcus pentosaceus. P. pentosaceus BCNU 9070 showed resistance to bile acids and acidic pH. The P. pentosaceus BCNU 9070 strain also inhibited the cell growth of six food-borne pathogens including Listeria monocytogenes and Shigella sonnei. In addition, the P. pentosaceus BCNU 9070 strain expressed bile salt hydrolase activity and showed an ability to assimilate cholesterol in vitro. On the basis of these results, P. pentosaceus BCNU 9070 is considered to have probiotic potential for applications in functional foodstuffs.
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