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Student Number 88224014
Author Chia-Jen Yang(楊嘉蓁)
Author's Email Address No Public.
Statistics This thesis had been viewed 2118 times. Download 11 times.
Department Life Science
Year 2000
Semester 2
Degree Master
Type of Document Master's Thesis
Language zh-TW.Big5 Chinese
Title Isolation of Triton X-100 degraders and characterization of key degrading oxygenase
Date of Defense 2001-07-18
Page Count 114
Keyword
  • bacteria identificat
  • biodegradation
  • surfactant
  • Abstract riton X-100, a nonionic surfactant, was used in industrial, agricultural, and domestic applications. This surfactant belongs to octylphenol polyethoxylates and is a mixture with an average numbers of 9.5 ethoxylate units in this mixture. In this study, twenty bacterial strains were isolated from soil samples from pesticide factory (green house and ditch), ricefield nearby NCU and its ditch, drainage of dormitory at NCU, drainage of pond of NCU by using Triton X-100 as sole carbon source. They were identified by BioLog® method and 16S rDNA sequences. Among them, except of Pseudomonas putida and Pseudomonas sp., Moraxella osloensis, Brevbacterium iodinum, Ochrobactrum anphropi, Alcaligenes eutrophus, Alcaligenes xylosoxidans and Ensifer adhaerens have not been reported to be capable in the degradation of Triton X-100. Brevbacterium iodinum was the only Gram-positive bacterium isolated in this study. Oxygen uptake activity by bacteria grown from MSB containing 0.5% Triton X-100 demonstrated that Pseudomonas sp. (82.10.6), name as Pseudomonas sp. SH4, showed the highest activity, specific activity was 528 mmole/min/kg of wet weight, and was 16.5 fold than Pseudomonas sp. strain TR01 which was reported in 1994 to degradation of Triton N-101 by Hideaki et al.
    Table of Content 目錄………………………………………………………………….. I
    表目錄………………………………………………………………..III
    圖目錄….………………………………………………………….…IV
    名詞縮寫對照表……………………………………………………..VI
    中文摘要……………………………………………………………..VII
    英文摘要……………………………………………………………..IX
    壹、緒論………………………………………………………….….1
    一、界面活性劑的定義、分類、性質與使用……………………..1
    二、界面活劑之微生物分解……………………………………..4
    三、Triton X-100代謝途徑…….…………………………………9
    四、Triton類代謝物之環境荷爾蒙效力………………………..10
    五、研究動機與大綱…………………………………………….11
    貳、材料與方法…………………………………………………….13
    一、分解菌之篩選……………………………………………….13
    二、分解菌之鑑定……………………………………………….15
    三、菌種生長特性…………….………………………………….19
    四、加氧酵素活性測定………………………………………….20
    五、LC/MS分析………….………………………………………22
    六、酵素活性最適化…………………..…………………………24
    七、酵素純化……………………..………………………………25
    八、酵素之初步定性……………………………………………..29
    九、化學藥品與儀器設備………………………………………..29
    參、結果…………………………………………………………….31
    一、分解菌之篩選與分離……………………………………….31
    二、菌種鑑定…………………………………………………….32
    三、Triton X-100代謝途徑…….………………………………..36
    四、Triton X-100加氧酵素活性測試與純化……………………38
    五、Triton X-100加氧酵素之初步生化特性….…………………42
    肆、討論……………………………………………………………..43
    伍、結論與建議………………………………………………………50
    陸、參考文獻………………………………………………………..52
    表……………………………………………………………………..59
    圖……………………………………………………………………..72
    附錄一、不同溫度之水溶氧量………………………………………114
    Reference 朝京寧。1993。界面活性劑相關資訊介紹。化工資訊。台灣。第84-92頁。
    賴耿陽 譯。1994。界面活性劑應用實務。復漢出版。台灣。第375-358頁。
    廖明隆 譯。1994。界面化學與界面活性劑。文原書局。台灣。第13-35頁。
    王鳳英 譯。1998。界面活性劑的原理與應用。高立圖書。台灣。第233-366頁。
    林美鳳。2000。建立苯環化合物分解菌中苯環加氧與切割?之基因偵測法。國立中央大學生命科學研究所碩士論文。

    Ahel, M., W. Giger, and C. Schaffner. 1994a. Behaviour of alkylphenol polyethoxylate surfactants in the aquatic environment. I. Occurrence and transformation in sewage treatment. Water Res. 28:1131-1142.
    Ahel, M., W. Giger, and C. Schaffner. 1994b. Behaviour of alkylphenol polyethoxylate surfactants in the aquatic environment. II. Occurrence and transformation in rivers. Water Res. 28:1143-1152.
    Cook, A. M., H. Laue, and F. Junker. 1999. Microbial desulfonation. FEMS Microbiol. Rev. 22: 399-419.
    Anonymous. 1995. Male reproductive health and environmental oestrogens. Lancet. 345:933-935.
    Ausubel, F. M., R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith, and K. Struhl. 1990. Current protocols in molecular Biology. John Wiley & Sons, Inc., Boston. pp. 2.4.1-2.4.2.
    Ball, H. A., M. Reinhard, and P. L. McCarty. 1989. Biotransformation of halogenated and nonhalogenated octylphenol polyethoxylate residues under aerobic and anaerobic conditions. Environ. Sci. Technol. 23:951-961.
    Bicknell, R. J., A. E. Herbison, and J. P. Sumpter. 1995. Oestrogenic activity of an environmentally persistent alkylphenol in the reproductive tract but not the brain of rodents. J. Steroid Biochem. Mol. Biol. 54:7-9.
    Blackburn, M. A., and M. J. Waldock. 1995. Concentrations of alkylphenols in rivers and estuaries in England and Wales. Water Res. 29:1623-1629.
    Bochner, B. R. 1989. Sleuthing out bacterial identities. Nature 339:157-158.
    Brunner, P. H., S. Capri, A. Marcomini, and W. Giger. 1988. Occurrence and behaviour of linear alkylbenzensulphonates, nonylphenol, nonylphenol mono- and nonylphenol diethoxylates in sewage and sewage sludge treatment. Water Res. 22:1465-1472.
    Burger, K . 1963. Methods for the microdetermination and trace detection of surfactant. III. Trace detection and determination of surface-active polyethoxylate compounds and polyethylene glycols. Z. Anal. Chem. 196:251-259.
    Caldwell, D. R. 1995. Catabolic metabolism. In microbial physiology and metabolism. Caldwell W. C. Brown Communications. Inc., USA. pp. 83-115.
    Cheng, C-L., S-H. Chou, and S-L. Huang. 2001. Personal communication.
    Chern, L-l. 1999. Phylogenetic approach of classification of the bacteria. CCRC News 6-9.
    Chou, S-H., and S-L. Huang. 2001. Personal communication.
    Corcia, A. D. 1998. Characterization of surfactants and their biointermediates by liquid chromatography-mass spectrometry. J. Chromatogr. 794:165-185.
    Crabb, N., and H. Persinger. 1968. A determination of the apparent molar absorption coefficients of the cobalt thiocyanate complexes of nonylphenol ethylene oxide adducts. J. Am. Oil Chem. 45:611-615.
    Crescenzi, C., A. D. Corcia, and R. Samperi. 1995. Determination of nonionic polyethoxylate surfactants in environmental waters by liquid chromatography/electrospray mass spectrometry. Anal. Chem. 67:1797-1804.
    Deqin, M., S. M. Beverley, and S. J. Turco. 1996. Leishmania donovani passess a NADPH-dependent alkylglycerol cleavage enzyme. Biochem. Biophys. Res. Commun. 227:885-889.
    Ding, W-H., and S-H. Tzing. 1998. Analsis of nonylphenol polyethoxylates and their degradation products in rive r water and sewage effluent by gas chromatography—ion trap (tandem) mass spectrometry with electron impact and chemical ionization. J. Chromatogr. A. 824:79-90.
    Dominic, M. J., and G. F. White. 1998. Mechanism for biotransformation of nonylphenol polyethoxylates to xezoesterogens in Pseudomonas putida. J. Bacteriol. 180:4332-4338.
    Ferguson, P L., C. R. Iden, and B. J. Brownawell. 2000. Analysis of alkylphenol ethoxylate metabolites in the aquatic environment using liquid chromatography-electrospray mass spectrometry. Anal. Chem. 72:4322-4330.
    Field, J. A., and R. L. Reed. 1996. Nonylphenol polyethoxy carboxylate metabolites of nonionic surfactant in U.S. paper mill effluents, municipal sewage treatment plant effluents, and river waters. Environ. Sci. Technol. 30:3544-3550.
    Frings, J., E. Schramm, and B. Schink. 1992. Enzyme involved in anaerobic polyethylene glycol degradation of Pelobacter venetianus and Bacteroides strain PG1. Appl. Environ. Microbiol. 58:2164-2167.
    Giger, W., P. H. Brunnen, and C. Schaffner. 1984. 4-nonylphenol in sewage sludge: accumulation of toxic metabolites from nonionic surfactants. Science 225:623-625.
    Ginkel van C. G. 1996. Complete degradation of xenobiotio surfactant by consortia of aerobic microorganisms. Biodegradation 7:151-164.
    Hanioka, N., H. Jinno, Y-S. Chung, T. Nishimura, T-K. Tanaka, and M. Ando. 2000. Effect of 4-tert-octylphenol on cytochrome P450 enzyme in rat liver. Arch. Toxicol. 73:625-631.
    Hanioka, N., T. Tanaka-Kagawa, Y-S. Chung, T. Nishimura, H. Jinno, and M. Ando. 2000. Changes in Hepatic cytochome P450 enzymes by biodegradation products of 4-tert-octylphenol polyethoxylate in rats. Bull. Environ. Contam. Toxicol. 64:804-810.
    Harayama, S., A. Wasserfallen, P. Cerdan, and M. Rekik. 1992. Mutation modification of the substrate specificity of catechol 2,3-dioxygenase encoded by TOL plasmid pWW0 of Pseudomonas putida. In American society for microbiology. E. Galli, S. Siliver, and B. Witholt(ed.). Washington, D. C. pp. 223-230.
    Hawrelak, M., E. Bennett, and C. Metcalfe. 1999. The environmental fate of the primary degradation products of alkylphenol ethoxylate surfactants in recycled paper sludge. Chemosphere 39:745-752.
    Hideaki, M., N. Masuda, Y. Fujiwara, M. Ike, and M. Fujika. 1994. Degradation of alkylphenol ethoxylates by Pseudomonas sp. strain TR01. Appl. Environ. Microbiol. 60:2265-2271.
    Holt, J. G., N. R. Krieg, P. A. Sneath, J. T. Staley, and S. T. Williams. 1994. Bergey’s Manual of determinative Bacteriology. A waverly company. Ninth edition.
    Hosea, N. A., and F. P. Guengerich. 1998. Oxidation of nonionic detergents by cytochome P450 enzymes. Arch. Biochem. Biophys. 353:365-373.
    Kamiusuki, T., T. Monde, F. Nemoto, T. Konakahara, and Y. Takahashi. 1999. Separation and characterization of octylphenol ethoxylate surfactants used by reversed-phase high-performance liquid chromatography on branched fluorinated silica gel columns. J. Chromatogr. A. 852:475-485.
    Kawai, F. 1985. Existence of ether bond-cleaving enzyme in polyethylene glycol-utilizing symbiotic mixed culture. FEMS Microbiol. Lett. 30:273-276.
    Kuo, L-C., and S-L., Huang. 2001. Personal communication.
    Kvestak, R., and M. Ahel. 1994. Occurrence of toxic metabolites from nonionic surfactants in the Krka river estuary. Ecotoxicol. Environ. Saf. 28:25-34.
    Lebaron, P., J. Ghiglione, C. Fajon, N. Batailler, and P. Normand. 1998. Phenotypic and genetic diversity within a colony morphotype. FEMS Microbiol. Let. 160:137-143.
    Masai, E., Y. Katayama, S. Nishikawa, M. Yamasaki, N. Morohoshi, and T. Haraguchi. 1989. Detection and localization of a new enzyme catalyzing the b-aryl ether cleavage in the soil bacterium (Pseudomonas paucimobilis SYK-6). FEBS Lett. 249:384-352.
    Masai, E., Y. Katayama, S. Kawai, S. Nishikawa, M. Yamasaki, and N. Morohoshi. 1991. Cloning and sequencing of the gene for a Pseudomonas paucimobilis enzyme that cleaves b-aryl ether. J. Bacteriol. 173:7950-7955.
    Masai, E., S. Kubota, Y. Katayama, S. Kawai, M. Yamasaki, and N. Morohoshi. 1993. Characterization of the Cα-dehydrogenase gene involved in the cleavage of b-aryl ether by Pseudomonas paucimobilis. Biosci. Biotech. Biochem. 57(10):1655-1659.
    Masai, E., Y. Katayama, S. Kubota, S. Kawai, M. Yamasaki, and N. Morohoshi. 1993. A bacterial enzyme degrading the model lignin compound b-etherase is a member of glutathione-S-transferase superfamily. FEBS Lett. 323:135-140.
    Maki, H., M. Fujia, and Y. Fujiwara. 1996. Identification of final biodegradation product of nonyphenol ethoxylate (NPE) by river microbial consortia. Bull. Environ. Contam. Toxicol. 57:881-887.
    Masai, E., Y. Katayama, S. Nishikawa, and M. Fukuda. 1999. Characterization of Sphingomonas paucimobilis SYK-6 genes involved in degradation of lignin-related compounds. J. Ind. Microbiol. Biotechnol. 23:264-373.
    Matthew, J. S, and N. J. Malcolm. 2000. The biodegradation of surfactants in the environment. Biochimica et Biophysica Acta. 1508:235-251.
    McInroy, S. G., C. D. Campbell, K. E. Haukka, D. W. Oedd, J. I. Sprent, W. Wang, J. Young, and J. M. Sutherland. 1999. Characterisation of rhizobia from African acacias and other tropical woody legumes using BioLog™ and partial 16S rRNA sequencing. FEMS Microbiol. Let. 170:111-117.
    MicrologTM system, Release 4.0 User Guide. 1998. Biolog. Inc., USA. Section 13. pp. 3-20.
    Nguyen, M. H. and J-C. Sigoillot. 1997. Isolation from coastal sea water and characterization of bacterial strains involved in non-ionic surfactant degradation. Biodegradation 7:369-375.
    Nielsen, W. L., C. Filipe, L. Grady, JR, S. Molin, and A. Stahl. 1999. Identification of a novel group of bacteria in sludge from a deteriorated Biological phosphorus removal reactor. Appl. Environ. Microbiol. 65:1251-1258.
    Nimrod, A. C., and W. H. Benson. 1996. Environmental estrogenic effects of alkylphenol ethoxylates. Crit. Rev. Toxicol. 26:335-364.
    Patoczka, J., and G.W. Jones. 1990. Biodegradation and secondary effluent toxicity of ethoxylated surfactants. Wat. Res. 24:965-972.
    Pearce, B. A., and M. T. Heydeman. 1980. Metabolism of di(ethylene glycol)[2-(2’-hydroxyethoxy)ethanol] and other short poly(ethylene glycol)s by gram-nedative bacteria. J. Gen. Microbiol. 118:21-27.
    Pedersen, S. N., and C. Lindholst. 1999. Quantification of xenoestrogens 4-tert.-octylphenol and bisphenol A in water and fish tissue based on microwave assisted extraction, solid-phase extraction and liquid chromatography-mass spectrometry. J. chromatogr. 864:17-24.
    Pennisi, E. 1998. Genome data shake tree of life. Science 280:672-674.
    Petrovic, M., and D. Barcelo. 2000. Determination of anionic and nonionic surfactants, their degradation products, and endocrine-disrupting compounds in sewage sludge by liquid chromatography/mass spectrometry. Anal. Chem. 72:4560-4567.
    Powlowski, J., and V. Shingler. 1994. Genetics and biochemistry of phenol degradation by Pseudomonas sp. strain CF600. Biodegradation 5:219-236.
    Sharpe, R. M., and N. E. Skakkebaek. 1993. Are oestrogents involved in falling sperm counts and disorders of the male reproductive tract? Lancet. 341:1392-1395.
    Smith, M. R. 1990. The biodegradation of aromatic hydrocarbons by bacteria. Biodegradation. 1:191-206.
    Sneath, P. H. A., N. S. Mair, M. E. Sharpe, and J. G. Holt. 1986. Bergey’s Manual of Systematic Bacteriology. Waverly co., Maryland.
    Stanier, R. Y. G., C. Bazire, and W. R. Sistrom. 1957. Kinetics studies of pigment synthesis by non-sulfur purple bacteria. J. Cell. Comp. Physiol.49:25.
    Strubel, V., K. H. Engesser, P. Fischer, and H. J. Knackmuss. 1991. 3-(2-hydroxyphenyl)catechol as substrate for proximal meta ring cleavage in dibenzofuran degradation by Brevibacterium sp. strain DPO 1361. J Bacteriol. 173(6):1932-7.
    Suen, W-C., and D. T. Gibson. 1993. Isolation and preliminary characterization of the subunits of naphthalene dioxygenase from Pseudomonas putida NCIB9816-4. J. Bacteriol. 175:5877-5881.
    Tanghe, T., W. Dhooge, and W. Verstraete. 1999. Isolation of a bacterial strain able to degrade branched nonylphenol. Appl. Environ. Microbiol. 65:746-751.
    Thélu, J., L. Medina, and J. Pelmont. 1980. Oxidation of polyoxyethylene oligomers by an inducible enzyme from Pseudomonas P400. FEMS Microbiol. Lett. 8:187-190.
    Watson, G. K., and N. Jones. 1977. The biodegradation of polyethylene glycols by sewage bacteria. Water Res. 11:95-100.
    White, G. F. 1993. Bacterial biodegradation of ethoxylated surfactants. Pestic. Sci. 37:159-166.
    White, G. F., N. J. Russell, and E. D. Tidswell. 1996. Bacterial scission of ether bonds. Microbiol. Rev. 60:216-232.
    White, R., S. Jobling, S. A. Hoare, J. P. Sumpter, and M. G. Parker. 1994. Environmentally persistent alkylphenolic compound are estrogenic. Endocrinology. 135:175-183.
    Zylstra, G. J., and D. T. Gibson. 1989. Toluene degradation by Pseudomonas putida F1. J. Biol. Chem. 264:14940-14946.
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