Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/28006
標題: Thermophilic Alkaline-Protease-Producing Bacterium - Tepidimonas taiwanensis sp.nov. from Southern Taiwan
南臺灣溫泉之嗜熱性鹼性蛋白酶生產菌Tepidimonas taiwanensis sp.nov.之研究
作者: 陳天來
Chen, Tien-Lai
關鍵字: Tepidimonas taiwanesis
Tepidimonas taiwanesis
β-Proteobacteria
Hot spring
Alkaline protease
Taxonomy
β-Proteobacteria
溫泉
鹼性蛋白酶分類學
出版社: 土壤環境科學系所
引用: 王美人,游若荻。1995。篩選枯草菌之DNA轉形株用於蛋白酶之生產。食品科學,22(1):38-45。 王昭閔。2000。好氣式堆肥處理之菌相調查及嗜高溫菌之分類。國立中興大學獸醫微生物學研究所碩士論文。 王進琦。1991。微生物學實驗。p.31-60。藝軒圖書出版社。臺灣,台北。 江啟銘。1995。Ⅰ、不同生長期及不同部位之豌豆植物中果膠脂酶同功酶之消長及其純化、生理意義及應用性之研究Ⅱ、果膠脂酶同功酶反應機制之研究。國立臺灣大學農業化學研究所博士論文。 呂鋒洲,林仁混。1987。基礎酵素學。聯經出版事業公司,台北。 沈佛亭。2006。Gordonia菌屬放線菌之分子偵測、分類及鑑定。國立中興大學土壤環境科學系博士論文。 林詠婷a。2000。臺灣烏來溫泉嗜熱性菌 Bacillus thermocatenulatus WL65之篩選。國立臺灣大學植物學研究所碩士論文。 林耀松b。2000。國家生物多樣性研究之規劃報告。行政院國科會生物處。 邱清祥。2005。臺灣溫泉中分離出嗜熱性澱粉酶新菌種。國立高雄海洋科技大學食品科學所碩士論文。 陳怡宏。1997。蛋白酵素水解液生產技術。食品工業,29(11):34-40。 陳懋彥。2002。臺灣地區嗜熱性細菌之研究。國立臺灣大學植物學研究所博士論文。 莊榮輝。1985。水稻蔗糖合成酶之研究。國立臺灣大學農業化學研究所博士論文。 黃卓治,辛志勳,張文重。1978。納豆菌之研究納豆菌蛋白質分解酵素生產條件之探討。屏東農專學報,19:92-98。 楊秋忠、劉瑞美、沈佛亭。2002。逢機複製多型性DNA技術分析溶磷 與不溶磷根瘤菌的基因親緣性。土壤肥料通訊 84:167-168。 楊秋忠。2003。微生物肥料在永續農業之應用。微生物之多樣性及其永 續利用研討會。國立臺灣大學植物病理與微生物學系。p. 48-54. 楊蘇聲。1997。細菌分類學。中國農業大學出版社。中國北京 蔡文城。2003。微生物學。藝軒圖書出版社。臺灣台北 賴威安。2000。Bacillus sp.P-6 中蛋白酶的生產與性質分析。國立中興大學食品科學系碩士論文。 Altas, R.M., and A.K. Bej. 1994. Polymerase Chain reaction. In Methods for general and molecular bacteriology. p. 418-435. ed. by P.Gerhardt.Washington, D.C. American society for Microbiology. Bednarski, W., and J. Kowalewska-Piontas. 1986. Thermostable enzymes in food technology. Postepy Biochem. 32:469-484. Benson, H. J. 2002. Microbiological Applications. p.84,138.New York:McGraw Hill. Benyon, R. J., and J. S. Bond. 1989. The Schecter and Berger nomenclature for protease subsites in “proteolytic enzyme”. p.231.ed. R. J.Beynon and J. S.Bond. IRL Press. Beveridge, T.J., T.J. Popkin, and R.M. Cole. 1994. Electron microscopy. In Methods for general and molecular bacteriology. p.665-682. ed. by P.Gerhardt. Washington, D.C. American Society for Microbiology. Bollag, D.M., M.D. Rozycki, and S.J. Edelstein. 1996. Protein methods. Wiley-Liss Inc, USA, New York. Breccia, J. D., F.M. Sinerize, D.G. Baigori, R. Castro., and H.R. Kaul. 1998. Purification and characterization of a thermostable xylanase from, Bacillus amyloliquefaciens. Enzyme Microb. Technol. 22:42-49 Brock,T.D. and H. Freeze. 1969. Thermus aquaticus gen.nov.and sp. nov., a nonsporulating extreme thermophile. J. Bacteriol. 98:289-297. Brock,T.D. 1978. Thermophilic Microorganisms and life at high temperatures. Springer-Verlag,New York. Brock,T.D. 1986. Thermophiles. General,Molecular and Applied Microbiology. John Wiley & Sons, New York. Brosius, J., M.L. Palmer, P.J. Kennedy, and H.R. Noller. 1978. Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli.Proc. Natl. Acad. Sci. U.S.A. 75:4801-4805. Chiang, W.D., C.J. Shih, and Y.H. Chu. 1999. Functional properties of soy protein hydrolysate produced from a continuous membrane reactor system. Chien, A., D.B. Edgar, and J.M. Trela. 1976. Deoxyribonucleic acid polymerase from the extreme thermophile Thermus aquaticus.J Bacteriol. 127:1550-1557. Chung,A.P., F. Rainey, M.F. Nobre, J. Burghardt, and M.S. da Costa. 1997. Meiothermus cerbereus sp. nov.,a new slightly thermophilic species with high levels of 3-hydroxy fatty acids. Int.J.Syst.Bacteriol.47:1225-1230. Chung,B.T. 1995. Study of hot springs and geothermal environments of Taiwan (in Chinese). Ener.Resour.Environ.Quart. 8:35-42. Claudia, C.H., and R.A. Finkelstein. 1993. Bacterial extracellular zinc-containing metaloproteases. Microbiol.Rev. 54:823-837. Collins, M.D. 1985. Analysis id isopronoik quinines. In Methods in microbiology. P.329-366. ed. by G.Gottschalk. London. Academic Press, New York. da Costa, M.S.1995. The cell walls and lipids of Thermus. In Thermus species. P.143-253. ed. by R.Sharp and R.A.D.Williams. Plenum Press, New York. Daniels, L., R.S. Hanson, and J.A. Philips. 1994. Chemical analysis. In Methods for general and molecular bacteriology. p.655-682. ed. by P. Gerhardt. Washington, D.C. American Society for Microbiology. Dorsch, M., and E. Stackebrandt. 1992. Some modifications in the procedure of direct sequencing of PCR amplified 16S rDNA. J. Microbiol. Methods. 16:271-279. Edwards, U., T. Rogall, J. Blocker, M. Emde, and E.C. Bottger. 1989. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res. 17:7843-7853. Eijsink,V.G.H., O.R. Veltman, W. Aukema, G. Vriend, and G. Venema. 1995. Structural determinants of the atability of thermolysin-like proteinases. Structural Biol.2:374-379. Felstein, J. 1989. PHYLIP-Phylogeny Inference Package. Cladistics. 5:164-166. Fox,G.E., J.D. Wisotzkey., and P.JR. Jurtshuk. 1992. How close is close:16S rRNA sequence identity may not be sufficient to guarantee species identity. Int.J.Syst.Bacteriol.42:166-170. Freitas M., F.A. Rainey, M.F. Nobre, A.J. Silvestre, and M.S. da Costa. 2003. Tepidimonas aquatica sp.nov.,a new slightly thermophilic beta-proteobacterium isolated from a hot water tank. Syst Appl Microbiol.26:376-381. Fujiwara, S., S. Okuyama, and T. Imanaka. 1996. The world of archaea:genome analysis,evolution and thermostable enzymes. Gene. 179:165-170. Funke, G., F.N.R. Renaud, J. Freney, and P. Riegel. 1997. Multicenter evaluation of the updated and extended API (RAPID) Coryne database 2.0.J. Clin. Microbiol. 35:3122-3126. Gherna,R.L.1994.Culture preservation. In Methods for General&Molecular Bacterialogy,pp.278-292. ed. by P.Gerhardt. Washington,D.C.:American Society for Microbiology. Gillis, M., P. Vandamme, P. De Vos, J. Swings, and K. Kersters. 2001. Polyphasic Taxonomy. In Bergey''s manual of systematic bacteriology Vol L.,p.43-48. ed. by D.R.Boone., R.W.Castenholz , and G.M.Garrity. Springer, New York. Goodfellow, M., and A.G. O''Donnell. 1994. Roots of bacterial systematics. In Handbook of new bacterial systematics. p.3-56. ed. by M.Goodfellow and A.G.O''Donnell. 2th ed.Academic Press, London. Hall,T.A. 1999. BioEdit:auser-friendly biological sequence alignment editor and analysis program for windows 95/98/NT.Nucl Acids Symp Ser 41:95-98. Hagihara, B., H. Matsubara, M. Nakai, and K. Okunki. 1958. Study on assay of proteolytic activity. J. Biochem. 45:185-194. Hartley, B.S. 1960. Proteolytic enzymes. Ann.Rev.Biochem.29:45. Henner, D.J., E. Ferrari, M. Perego, and A. Hoch. 1988. Location of the targets of the npr-97, sac U32(Hy) and sac Q36(Hy) mutations in the upstream regions of the subtilisin promoter. J. Bacteriol. 170:296-300. Holt, J.G., and N.R. Krieg. 1994. Enrichment and isolation. In Methods for general and molecular bacteriology, p.179-215. ed. by P.Gerharat.Weshington, D.C. American Society for Microbiology. Hoppert, M., and A. Holzenburg. 1998. Electron Microscopy in Microbiology. Guildford, UK: BIOS Scientific Publisher. Hyun, H.H., and J.S.Zeikus. 1985. General biochemical characterization of thermostable extra cellular β-amylase from clostridium thermosulfurogenes. Appl. Environ. Microbio.49(5.) Imanaka, T. 1994. Thermostable enzymes produced by recombinant mesophilic and thermophilic bacteria. Bioprocess Technol 19:301-309. Johnson, W. C., and A. J. Lindsay. 1939. An improved universal buffer. Analyst. 64:958-993. Johnson, J.L. 1984. Nucleic acids in bacterial classification. In Bergey’s Manual of Systematic Bacteriology. p.8-11. ed. by N.R.Krieg. and J.G.Holt. Baltimore,MD. Walliams & Wilkins. Johnson, J.L. 1985. Determination of DNA base composition. In Methods in Microbiology,p.1-32. ed. by G.Gottschalk. London; Academic Press, New York. Johnson, J.L. 1994. Similarity analysis of DNAs. In Methods for General and Molecular bacteriology. p.655-682. ed. by P.Gerharat.Weshington, D.C. American Society for Microbiology. Jones, B.L., D. Fontanini, M. Jarvinen, and A. Pekkarinen. 1997. Simplified endoproteinase assays using gelatin or azogelatin. Anal. Biochem. 263:214-220. Jukes, T.H., and C.R. Cantor, 1969. Evolution of protein molecules. In Mammalian Protein Metabolism, p.21-132. ed. by M.H.N. Academic Press, New York. Krieg, N.R. 2001. Identification of Procaryotes. In Bergey’s manual of systematic bacteriology Vol I.,p.33-38. ed. by D.R.Boone., R.W.Castenholz. and G.M. Garrity. Springer, New York. Kristjansson, J.K., and G.A. Alfredsson.1992a. Thermophilic bacteria. CRC Press, Boca Raton. Kristjansson, J.K., and G.A. Alfredsson. 1992b. The heterotrphic thermophilic genera Thermomicrobium, Rhodothermus, Saccharococcus, Acidothermus and Sacotothermus. In Thermophilic Bateria.pp.64-76. ed. by J.K.Kristjansson. CRC Press, Boca Raton. Kristjansson, J.K., S. Hjorleifsdottir, V.T. Marteinsson, and G.A. Alfredsson. 1994. Thermus scotoductus, sp. Nov., a pigmented-producing thermophilic bacterium from hot tap water in Iceland and including Thermus sp. X-1. Syst. Appl. Microbiol. 17:44-50. Kumar, C.G., and T H. Takagi, 1999. Microbial alkaline proteases: from a bioindustrial viewpoiont. Biotechnol.Adv.17:561-594. Kuykendall, L.D., M.A. Roy, J.J. O’Neill, and T.E. Devine. 1988. Fatty acids, antibiotic resistance, and deoxyribonucleic acid homology groups of Bradyrhizobium japonicum. Int. J. Syst. Bacteriol. 38:358-361. Ladrat, C., L. Cornec, A.M. Alayse-Danet, and G. Barbier. 1995. Demonstration of thermostable enzymes in thermophilic micro-organisms of hydrothermal origin.C R Acad Sci Ⅲ 318,423-429. Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680-685. Lahl, W.J., and S.D. Braun. 1994. Enzymatic production protein hydrolysates for food use.Food Technol.48(10):68-71 Lane, D.J. 1991. 16S/23S rRNA sequencing. In Nucleic acid techniques in bacterial systematics. p.115-175. ed. by Stackebrandt E., and M. Goodfellow. Chichester, John Wiley. Lin, L.L., M.R. Tsau, and W.S. Chu. 1996. Purification and properties of a 140 kDa amylopullulanase from thermophilic and alkaliphilic Bacillus sp.strain TS-23. Biotechnol. Appl. Biochem.24:101-107. Lin, X., C.G. Lee, E.S. Casale, and J.C.H. Shih. 1992. Purification and characterization of a keratinase from a feather-degrading Bacillus licheniformis strain. Appl. Environ . Microbiol. 58(10):3271-3275. Loffler, A. 1986. Proteolytic enzymes:sources and applications. Food Technol. 40(12):63-70. Ludwig, W., and H. Klenk. 2001. Overveiw:A phlogenetic backbone and taxomic framework for procayotic systematics. In Bergey’s Manual of Systematic Bacteriology Vol Ⅰ,2ed., p.43-48. ed. by Boone, D.R., and R.W. Castenholz. Springer-Verlag, New York. Madigan, M.T., and B.L. Marrs. 1997. Extremophiles. Sci,Am. 276, 82-87. Maidak, B.L., N. Laresn, N.J. McCaughey, R. Overbeek, G.J. Olsen, K. Gogel, J. Blandy, and C.R. Woese. 1994. The RibosomalDatabase Project.Nucleic Acids Reserch.22:3485-3487. Matta, H., and V. Punj. 1998. Isolation and partial characterization of a thermostable extracellular protease of Bacillus polymyxa B-17. Int. J. Food. Microbiol. 42: 139-145. Mesbah, M., and W.B. Whitman. 1989. Measurement of deoxyguanosine/thymidine ratios in complex mixtures by high-performance liquid chromatography for determination of the mole percentage guanine+cytosine of DNA.J Chromatogr. 479:297-306. Moreira, C., F.A. Rainey, M.F. Nobre, M.T. da Sliva, and M.S. da Costa. 2000. Tepidimonas ignava gen.nov.,sp.nov.,a new chemolithoheterotrophic and slightly thermophilic menber of the beta-Proteobacteria.Int J Syst Evol Microbiol. 50:735-742. Morgan, J.A.W., and C. Winstanley. 1997. Microbial biomarker. In Modern soil microbiology. P.331-352. ed. by van Elsas, J.D., J.T. Trevors, and E.M.H.Wellington. Marcel Dekker, New York, USA. Murray, R.G.E., and C.F. Robinow. 1994. Light microscopy. In Methods for General and Molecular Bacteriology. p. 655-682. ed. by P. Gerhardt.Washington, D.C. American Society for Microbiology. Nduwimana, J., L. Guenet, I. Dorval, M. Blayau, J. L. Gall, and A. L. Treut. 1995. Rroteases. Ann. Biol. Clin.53:251-264. Nichols, D.S., and T.A. McMeekin. 2002. Biomarker techniques to screen for bacteria that produce polyunsaturated fatty acids. J. Microbiol. Methods. 48:161-170. Nikolausz, M., R. Sipos, S. Revesz, A. Szekely, and K. Marialigeti. 2005. Observation of bias associated with re-amplification of DNA isolated from denaturing gradient gels. FEMS Microbiol. Lett. 244:385-390. Nishimura,T., and H. Kato. 1988. Taste of free amino acids and peptides. Food Rev.Int.4:175-194. Oshima, T. 1973. Thermostable enzymes of thermophiles and thermal stability of enzyme protein. Tanpakushitau Kakusan Koso.18:454-458. Pantazaki, A.A., A.A. Pritsa, and D.A. Kyriakides. 2002. Biotechnologically relevant enzymes from Thermus thermophilus. Appl Microbiol Biotechnol.58:1-12. Pantazaki, A.A., A.A. Karagiorgas, M. Liakopoulou-Kyriakides, and D.A. Kyriakidis. 1998. Hyperalkaline and thermostable phosphatase in Thermun thermophilius.Appl Biochem Biotechnol.75:249-259. Palys, T., L.K. Nakamura, and F.M. Cohan. 1997. Discovery and classification of ecological diversity in the bacterial word:the role of DNA sequence data. Int. J. Syst. Bacteriol. 47:1145-1156. Parvaresh, F., G. Vic, D. Thomas, and M.D. Legoy. 1990. Uses and potentialities of thermostable enzymes.Ann N Y Acad Sci. 613:303-312. Price, N.C. and L. Steven. 1999. Fundamentals of enzymology. p.184-185. Oxford university Press Inc, New York. Rainey, F.A., N. Ward-rainey, R.M. Kroppenstedt, and E. Stackebrandt. 1996. The genus Nocardiopsis represents a phylogenetically coherent taxon and a distince actinomycete lineage: proposal of Nocardiopsaceae fam. Nov. Int. J. Syst. Bacteriol. 46:1088-1092. Rainey, F.A., M.F. Nobre, P. Schumann, E. Stackebrandt, and M.S. da Costa. 1997. Phylogenetic diversity of the deinococci as determined by 16S ribosomal DNA sequence comparison. Int. J. Syst. Bacteriol. 47:510-514. Rao, M. B., A. M. Tanksale, M. S. Chatge, and V. V. Deshpande. 1998. Molecular and biotechnological aspects of microbial proteases. Microbial. Mol. Biol. Rev. 62(3):597-635. Ronimus, R.S., L.E. Parker, and H.W. Morgan. 1997. The utilization of RAPD-PCR for identifying thermophilic and mesophilic Bacillus species.FEMS Microbiol.Lett.145:75-79. Saitou, N., and M. Nei. 1987. The neighbor-joiming method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 4:406-425. Schaeffer,P. 1969. Sporulation&the production of antibiotics, exoenzymes and exotoxins Bacterial. Rev. 33:48-71 Silva, M.T., and J.C. Sousa. 1973. Ultrastructure of the cell wall and cytoplasmic membrane of gram-negative bacteria with different fixation techniques. J. Bacteriol. 113:953-962. Sohl, J.L., A.K. Shiau, S.D. Rader, B.J. Wilk, and D.A. Agard. 1997. Inhibition of α-lytic protease by pro region C-terminal steric occlusion of the active site. Biochemistry. 36:3894-3902. Spurr, A.R. 1969. A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26:31-43. Stackebrandt, E., and B.M. Goebel. 1994. A place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int. J. Syst. Bacteriol. 44:846-849. Stackebrandt, E., and W. Liesack. 1994. Nucleic acids and classification. In Handbook of new bacterial systematics. p.152-194. ed. by Goodfellow, M., and A.G. O''Donnell. 2th ed. Academic Press,London. Stefanitsi, D., G. Sakellaris, and J.R. Garel. 1995. The presence of two proteinases associates with the cell wall of Lactobacillus bulgaricus.FEMS Microbiol. Lett.1995.128:53-58. Suzuki, K., M. Goodfellow, and A.G. O''Donnell. 1994. Cell envelopes and classification. In Handbook of new bacterial systematics. p.195-250. ed. by Goodfellow,M. and A.G.O''Donnell. 2th ed.Academic Press,London. Tindall, B.J. 1989a. Fully saturated menaquinones in the archacbacterium pyrobaculum islandicum. FEMS Microbiol. Lett. 60:251-254. Tindall, B.J. 1989b. Lipid composition of Rhodothermus marinus. FEMS Microbiol. Lett. 80:65-68. Tindall, B.J., G.A. Tomlinson, and L.I. Hochstein. 1987. Polar lipid composition of a new halobacterium. Syst. Appl. Microbiol. 9:6-8. Tornabene, T.G. 1985. Lipid analysis and the relationship to chemotaxonomy. In Methods in Microbioogy. p.209-231. ed. by G.Gottschalk. London. Academic Press, New York. Vandamme, P., B. Pot, M. Gills, P. de vos, K. Kerster, and J. Swings. 1996. Polyphasic taxonomy, a consensus approch to bacterial systematics. Microbiol.Rev. 60:407-438. Vauterin, L., J. Swing, and K. Kersters. 1994. Protein electrophoresis and classification. In Handbook of new bacterial systematics. p.251-281. ed. by Goodfellow,M. and A.G.O''Donnell.2th ed.Academic Press, London. Wang,J.J., and J.C.H. Shih. 1999. Fermentation production of keratinase from bacillus licheniformis DWD-1 and a recimbinant B.subtilis FDB-29. J. Indust. Microbiol. Biotech. 22:608-616. Watts, D., and J.R. MacBeath. 2001. Automated fluorescent DNA sequencing on the ABI PRISM 310 Genetic Analyzer. Methods Mol. Biol. 167:153-170. Welch, D.F. 1991. Applications of cellular fatty acid analysis Clin. Microbiol. Rev. 4:422-438. Williams, C.M., C.S. Richter, J.M. Mackenzie, J.R., and J.C.H. Shih. 1990. Isolation,identification,and characterization of a feather-degrading bacterium. Appl. Environ. Microbiol. 56:1509-1515. Woese, C.R., and G.E. Fox. 1977. Phylogenetic structure of the prokaryotic domain: the primary kingdoms. Proc. Natl. Acad. Sci. U.S.A. 74:5088-5090. Woese, C.R. 1987. Bacterial evolution. Microbiol.Rev. 51:221-271. Woese, C.R., O. Kandler, and M.L. Wheelis. 1990. Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc. Natl. Acad. Sci. U.S.A. 87:4576-4579. Young, C. C., T. C. Lin, M. S. Yeh, F. T. Shen, and J. S. Chang. 2005. Identification and kinetic characteristics of an indigenous diesel-degrading Gordonia alkanivorans strain. World J. of Microbiol. Biotech.21: 1409-1414.
摘要: During screening for protease-producing bacteria, a strain designated Ⅰ1-1T was collected from a Sih-Chong-Si hot spring at Pingtung, located in the southern Taiwan. This strain formed light-yellow pigmented colonies. Cells of this organism were Gram-negative rods motile by means of a single polar flagellum. The sizes of rods were approximately 0.4-0.5 μm in diameter and 0.8-2.0 μm in length. Strain Ⅰ1-1T grew at temperatures ranging from 35 to 60℃, pH values between 6.0 and 10, and NaCl concentrations between 0-0.5%. Optimal growth condition was around 55℃, pH 7-8 and 0% NaCl. Comparison of the 16S rRNA gene sequence similarity of strainⅠ1-1T with those collected in public databases reveals that strain Ⅰ1-1T belonged to the comamonadaceae within Burkholderiales group of the β-Proteobacteria. According to the results from the phylogentic analysis, the 16S rDNA sequence similarty of strain Ⅰ1-1T clustered together with genus Tepimonas which contained species, On the basis of DNA-DNA similarity data, physiological and biochemical characteristics of API, Biolog, antibiotic susceptibility, fatty acid compositions, whole cell protein profile comparisons, as well as G+C content, the organism belonged to the protease genus Tepimonas and represented a novel species within this genus. We propose to classify Ⅰ1-1T strain as Tepidimonas taiwanesis sp.nov. The 16S rDNA sequence of strain Ⅰ1-1T clustered together with T. ignava(97.2﹪similarity)and T. aquatica.(96.8﹪similarity). Whole-genome DNA-DNA hybridization experiment was performed between strain Ⅰ1-1T and two type strains of it's nearest phylogenetic neighbors, T. ignava and T. aquatica strain. The binding levels of strain Ⅰ1-1T twords T. ignava and T. aquatica were 38.3﹪and 40.8﹪, respectively. The whole-cell protein profile of strain Ⅰ1-1T was compared with that of T. ignava(92.1﹪similarity) and T.aquatica (89.8﹪similarity). The G+C content of it's DNA is 68.1mol﹪. The predominant cellular fatty acids of strain Ⅰ1-1T were 16: 0 ( about 41﹪), 18:1 ω7 c ( about 13﹪), and summed feature 3a (16:1ω or 15:0 iso 2OH or both) (about 26﹪). A zymogram analysis of the strain Ⅰ1-1T in the alkaline medium reveales the presence of protease activity near the molecular weights of 32, 60, and 75 kDa. The condition of protease activity from the strain I1-1T in this research were assayed, and the optimal temperature was at 55℃, and optimal pH was at 8.
本研究之菌種來自南臺灣屏東地區四重溪溫泉水中之一株具分解蛋白能力的好氧性嗜熱桿菌,名為Ⅰ1-1T 菌株。Ⅰ1-1T 菌株經培養後菌落顏色呈現半透明淡黃色,菌體形態大小為0.4-0.5μm x 0.8-2.0μm之革蘭氏陰性菌,具單鞭毛運動能力。Ⅰ1-1T 菌株生長溫度範圍為35-60 ℃( 最適55℃),生長pH值範圍為6-10(最適pH7-8)‚生長鹽度範圍為0-0.5%的NaCl。Ⅰ1-1T 菌株經由16S rDNA 序列分析與資料庫比對,Ⅰ1-1T 菌株屬於β-Proteobacteria 綱、Burkholderiable 目、Comamonadaceae 科。經由系統分類學分析,確認Ⅰ1-1T 菌株與Tepidimonas 菌屬中的菌種之16S rDNA 序列相似度,再經由DNA-DNA 雜合反應、生化分析之API 系統、Biolog 系統、抗生素感受性、脂肪酸組成、全細胞蛋白質電泳、G+C content分析比對結果, Ⅰ1-1T 菌株屬於Tepidimonas 菌屬中的另一新種,因此命名為Tepidimonas taiwanesis。 Ⅰ1-1T 菌株與Tepidimonas 菌屬中之二菌種T. ignava 及T. aquatica 之16S rDNA 序列相似度分別為97.2﹪及96.8﹪,而Ⅰ1-1T 菌株與T. ignava 及T. aquatica DNA-DNA 雜合結果,其相似度分別為38.3﹪、40.8﹪。另全細胞蛋白質電泳結果Ⅰ1-1T 菌株與T. ignava 、T. aquatica 之序列相似度分別為92.1﹪、89.8﹪。此外, Ⅰ1-1T 菌株脂肪酸成分以16:0 ( 約41﹪)、18:1 ω7 c ( 約13﹪)和summed feature 3a (16:1ω or 15:0 iso 2OH or both) ( 約26﹪)為主。以Ⅰ1-1T菌株之胞外粗酵素液進行SDS-PAGE電泳分離,證明Ⅰ1-1T菌株具有在鹼性下生產蛋白酶的能力,其蛋白酶分子量約為32、60與75 kDa。本研究菌株粗酵素液的蛋白酶活性最適反應溫度為55℃ ,而最適作用pH值為8之鹼性條件。
URI: http://hdl.handle.net/11455/28006
其他識別: U0005-0702200720550100
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-0702200720550100
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