Isolation and Charaterization of Myxobacteria with Proteolytic Activity
Myxobacteria의 Proteolytic Activity 특성
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Seven isolates showing strong proteolytic activity, KYC 1028, 1100, 1134, 1139, 1151, 1159, and 1182, were collected. Out of them, the broth of KYC 1134 and KYC 1139 showed the high proteolytic activity measured by azocazein. To determine 16S rDNA sequences for identification, 16S rDNA of seven isolates were amplified and compared with the 16S rDNA sequences of other myxobacteria at NCBI. It is evident from the phylogenetic tree that the isolates belong to the genus Myxococcus. Sharing high percentage similarity values with myxobacteria, the 16S rDNA sequences were involved in two species, Myxococcus macrospores and M. Fulvus. Biochemical characteristics of KYC 1134 broth, which showed the highest proteolytic activity, showed increased activity 8 times to seven days after culture, and protein production were increased gradually and stopped at five days. The broth had optimal temperature at 60℃ for proteolytic activity, and stability of pH was ranged from pH 5 to 10, at 50℃ and 60, respectively. To classify proteases being in the broth, ten inhibitors were determined and only bestatin showed 27% inhibition effect. The inhibition result demonstrates that the broth contains kinds of amino peptidases and other exopeptidases.
Abstract
Seven isolates showing strong proteolytic activity, KYC 1028, 1100, 1134, 1139, 1151, 1159, and 1182, were collected. Out of them, the broth of KYC 1134 and KYC 1139 showed the high proteolytic activity measured by azocazein. To determine 16S rDNA sequences for identification, 16S rDNA of seven isolates were amplified and compared with the 16S rDNA sequences of other myxobacteria at NCBI. It is evident from the phylogenetic tree that the isolates belong to the genus Myxococcus. Sharing high percentage similarity values with myxobacteria, the 16S rDNA sequences were involved in two species, Myxococcus macrospores and M. Fulvus. Biochemical characteristics of KYC 1134 broth, which showed the highest proteolytic activity, showed increased activity 8 times to seven days after culture, and protein production were increased gradually and stopped at five days. The broth had optimal temperature at 60℃ for proteolytic activity, and stability of pH was ranged from pH 5 to 10, at 50℃ and 60, respectively. To classify proteases being in the broth, ten inhibitors were determined and only bestatin showed 27% inhibition effect. The inhibition result demonstrates that the broth contains kinds of amino peptidases and other exopeptidases.
참고문헌 (19)
- Bollag, D.M, "Epothilones, a new class of microtubule-stabilizing agents with Taxol-like mechanism of action", Cancer Res., 55, pp.2325~2333 (1995)
- Bradford, M. M, "A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding", Anal. Biochem, 72, pp.248~254 (1976)
- Gass. J, "Fermentation, Purification, Formulation, and Pharmacological Evaluation of a Prolyl Endopeptidase from Myxococcus Xanthus: Implication for Celiac Sprue Therapy", Biotechnology and Bioengineering, 92, pp.674~684 (2005)
- Gerth, K, "The soraphens: a family of novel antifungal compounds from Sorangium cellulosum (Myxobacteria). I. Soraphen A1 alpha: fermentation, isolation, biological properties", J. Antibiot, 47, pp.23~31 (1994)
- Gerth, K, "Myxobacteria: proficient producers of novel natural products with various biological activities-past and future", J. Biotechnol, 106, pp.233~253 (2003)
- Godfrey, T, "Industrial enzymology, 2nd ed.", pp.3~ (1996), Macmillan Publishers Inc.
- Irschik, H, "Chivosazol A, a new inhibitor of eukaryotic organisms isolated from myxobacteria", J. Antibiot, 48, pp.962~966 (1995)
- Jansen, R, "Disorazoles, highly cytotoxic metabolites from the Sorangicin-producing bacterium Sorangium cellulosum, strain So ce12", Liebigs Ann. Chem., 8, pp.759~773 (1994)
- Kimura, M, "A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences", J. Mol. Evol., 16, pp.111~120 (1980)
- Kumar, S, "Integrated software for molecular evolutionary genetics analysis and sequence alignment", Brief Bioinfor., 5, pp.150~163 (2004)
- Perlova, O, "Reconstitution of the Myxothiazol Biosynthetic Gene Cluster by Red", ET Recombination and Heterologous Expression in Myxococcus Xanthus, 2006, pp.72, 7485~7494
- Poza, M, "Cloning and Expression of clt Genes Encoding Milk-Clotting Proteases from Myxococcus xanthus 422", Appl Environ Microbiol, 70, pp.6337~6341 (2004)
- Rao, M. B, "Molecular and biotechnological aspects of microbial proteases", Mirobiology and Molecular Biology Review, 62, pp.597~635 (1998)
- Reichenbach, H, "The Myxococcales. in: Bergey’s Manual of Systematic Bacteriology", pp.1059~1143 (2004), Springer-Verlag
- Reichenbach, H, "The myxobacteria. in: The Procaryotes, 2nd ed.", pp.3416~3478 (1992), Springer Verlag
- Saitou, N, "The neighbor-joining method: a new method for reconstructing phylogenetic trees", Mol Biol Evol, 4, pp.406~425 (1987)
- Sarath, G, "Protease assay Method. in: proteolytic enzyme: a practical approach", pp.25~55 (1989), IRL Press
- Shan, L, "Comparative biochemical analysis of three bacterial prolyl endopeptidases", Biochem J., 383, pp.311~318 (2004)
- Thompson, J. D, "The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools", Nucleic Acids Res., 25, pp.4876~4882 (1997)
