博碩士論文 88224008 詳細資訊




以作者查詢圖書館館藏 以作者查詢臺灣博碩士 以作者查詢全國書目 勘誤回報 、線上人數:79 、訪客IP:3.234.214.113
姓名 林奕成( Yi-Chen Lin)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 陰離子界面活性劑sodium dodecylbenzene sulfonate分解菌篩選與脫磺酸酵素研究
(Isolation of a bacterium degrading sodium dodecylbenzene sulfonate and characterization of desulfonation enzyme)
相關論文
★ 鄰苯二酚加氧酵素的熱穩定性提昇研究★ Triton X-100 分解菌之分離和分解酵素之特性研究
★ Triton X-100加氧酵素之純化與定性★ Lactobacillus reuteri於酸性與膽鹽環境中之蛋白質體研究
★ 蕃茄根部受銅逆境之基因調控★ Pseudomonas nitroreducens TX1 異化辛基苯酚聚氧乙基醇之功能性蛋白質體學:以二維電泳法分析等電點4-8之蛋白質表現
★ Pseudomonas nitroreducens TX1之具耗氧活性之麩胺酸合成酶之單離★ 人類細胞株生產含多種亞型的 干擾素-a之蛋白質體學研究
★ 辛基苯酚之分解:分解菌和生物復育之菌相研究★ 分解辛基苯酚聚氧乙基醇之耗氧酵素(二氫硫辛醯胺脫氫酶)的純化與定性
★ AtNPR1轉殖番茄之性狀分析及抗病機制研究★ Pseudomonas putida TX2分解辛基苯酚聚氧乙基醇及其具雌激素活性代謝物之研究
★ 以功能性蛋白質體學研究Pseudomonas nitroreducens TX1生長於辛基苯酚聚氧乙基醇之代謝與逆境反應★ 以功能性蛋白質體學研究Pseudomonas putida TX2生長於 辛基苯酚聚氧乙基醇與辛基苯酚之代謝與逆境反應
★ 以功能性基因體學研究細菌異化辛基苯酚 聚氧乙基醇及抗逆境之基因★ Pseudomonas nitroreducens TX1中二氫硫辛醯胺脫氫酶分解辛基苯酚聚氧乙基醇之研究
檔案 [Endnote RIS 格式]    [Bibtex 格式]    [相關文章]   [文章引用]   [完整記錄]   [館藏目錄]   至系統瀏覽論文 ( 永不開放)
摘要(英) Linear alkylbenzene sulfonates (LAS) are the most widely used anionic surfactants today. These compounds accumulate in the environment and show toxic effects on organisms in polluted water. This strain utilized sodium dodecylbenzene sulfonate (DBS) as sole source of sulfur in a minimal salts medium containing 0.3 % succinate. The bacterium can’t be identified by BioLog method in Gram-negative database. Further identification by 16S rDNA sequence was performed and showed 98 % identity to many Enterobacter species. Phylogenetic tree was constructed from an alignment of 1,193 nucleotides of 16S rDNA sequences showing the highest phylogenic relationship with Enterobacter agglomerans. And the bacterium was designated Enterobacter sp. SH3. The growth of strain was demonstrated as a function in the decreasing of DBS, which was analyzed by liquid chromatography-mass spectroscopy (LC-MS). It’s specific that Enterobacter. sp. SH3 can utilize several alkanesulfonates and aromatic sulfonates as sole sulfur source to grow. The action of the release of sulfite from DBS was measured using Ellman’s reagent followed by the absorbance increased at 430 nm as an enzyme assay. The highest enzyme activity was observed in the addition of 500 mM NADPH and 3 mM FMN in 0.3 ml 10 mM Tris-HCl, pH 9.0. The preliminary biochemical feature of the enzyme acting in Enterobacter sp. SH3 was showed to be similar to an FMNH2-dependent alkanesulfonate monooxygenase from Escherichia coli EC1250. A series of column chromatography was applied to purify the desulfonation enzyme. The active fraction from chromatofocusing chromatography was at pI of 5.18-4.12. The native molecular weight was at the range of 60-150 kDa by Sephacryl S-200 gel filtration chromatography. This highly purified desulfonation enzyme pool (240 mg) was further separated by 2D-gel electrophoresis and identified by MALDI-TOF
關鍵字(中) ★ 界面活性劑
★ 生物分解
★ 菌種鑑定
★ 脫磺酸
關鍵字(英) ★ surfactant
★ biodegradation
★ bacterium identifica
論文目次 目錄------------------------------------------------------------------------------I
表目錄---------------------------------------------------------------------------III
圖目錄---------------------------------------------------------------------------IV
縮寫與全名對照表------------------------------------------------------------VII
中文摘要------------------------------------------------------------------------VIII
英文摘要------------------------------------------------------------------------IX
壹、緒論-------------------------------------------------------------------------1
一、前言---------------------------------------------------------------------1
二、界面活性劑的性質與分類------------------------------------------2
三、直鏈烷基苯磺酸鹽的生物轉化與分解---------------------------4
四、磺酸類化合物與細菌生長硫源的關係---------------------------5
五、加氧酵素與脫磺酸反應---------------------------------------------5
六、高效液相層析質譜儀分析界面活性劑---------------------------7
七、研究背景與目的------------------------------------------------------8
貳、材料與方法----------------------------------------------------------------11
一、界面活性劑分解菌的篩選------------------------------------------11
二、菌體培養條件之探討------------------------------------------------15
三、DBS利用菌的鑑定--------------------------------------------------17
四、界面活性劑的微生物轉化鑑定------------------------------------21
五、脫磺酸酵素之分析---------------------------------------------------24
六、藥品---------------------------------------------------------------------28
七、儀器設備---------------------------------------------------------------28
參、結果-------------------------------------------------------------------------30
一、菌株的篩選、鑑定與生化分析-----------------------------------30
二、生長測試--------------------------------------------------------------32
三、菌體生長與DBS之消耗------------------------------------------34
四、Enterobacter sp. SH3對TS的降解-------------------------------34
五、脫磺酸酵素的純化--------------------------------------------------35
肆、討論-------------------------------------------------------------------------38
一、DBS利用菌株的分離與鑑定--------------------------------------38
二、Enterobacter sp. SH3的生長特性探討---------------------------39
三、Enterobacter sp. SH3對DBS、TS的生物轉化-----------------40
四、脫磺酸酵素分析-----------------------------------------------------40
伍、結論與建議----------------------------------------------------------------43
陸、參考資料-------------------------------------------------------------------44
柒、表----------------------------------------------------------------------------50
捌、圖----------------------------------------------------------------------------57
參考文獻 Ainsworth, S. J. 1996. Soaps and detergents. Chem. Engs. News. 74: 32-54
Alexander, M. 1973. Nonbiodegradable and other recalcitrant molecules. Biotechnol. Bioeng. 15: 611-647
Argese, E., A. Marcomini, P. Miana, C. Bettiol and G. Perin. 1994. Submitochondrial particle response to linear alkylbenzene sulfonates, nonylphenol polyethoxylates and their biodegradation derivatives. Environ. Toxicol. Chem. 13: 737-742
Berth, P. and P. Jeschke. 1989. Consumption and fields of application of LAS. Tenside Surfactants Deterg. 26: 75-79
Bird, J. A. and R. B. Cain. 1972. Metabolism of linear alkylbenzene sulphonates by a Vibro sp. Biochem. J. 127: 46p
Bradford, M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254
Burdzy, G. and D. Hulanicka. 1981. Use of gene fusions to study expression of cysB, the regulatory gene of the cysteine regulon.
J. Bacteriol. 147: 744-751
Christopher, C., R. Allen, D. R. Boyd, F. Hempenstall, M. J. Larkin and N. D. Sharma. 1999. Contrasting effects of a nonionic surfactant on the biotransformation of polycyclic aromatic hydrocarbons to cis-dihydrodiols by soil bacteria. Appl. Environ. Microbiol. 65: 1335-1339
Chou, S.-H. and S.-L. Huang. 2001. Personal communications.
Cook, A. M., H. Laue, F. Junker. 1999. Microbial desulfonation. FEMS Microbiol. Rev. 22: 399-419
Cook, A. M. 1998. Sulfonated surfactants and related compounds: facets of their desulfonation by aerobic and anaerobic bacteria. Tenside. Surfact. Deterg. 35: 52-56
Cook, A. M. and R. Hutter. 1982. Ametryne and prometryne as sulfur sources for bacteria. Appl. Environ. Microbiol. 43: 781-786
Corcia, A. D. 1998. Characterization of surfactants and their biointermediates by liquid chromatography-mass spectrometry. J. Chromatogr. A. 794: 165-185
Dudley, M. W. and J. W. Frost. 1994. Biocatalytic desulfurization of arylsulfonates. Bioorg. Med. Chem. 2: 681-690
Eichhorn, E., J. R. van der Ploeg, and T. Leisinger. 1999. Characterization of a two-component alkanesulfonate monooxygenase from Escherichia coli. J. Biol. Chem. 274: 26639-26646
Fantroussi, S. E., L. Verschuere, W. Verstraete and E. M. Top. 1999.
Effect of phenylurea herbicides on soil microbial communities estimated by analysis of 16S rRNA gene fingerprints and community-level physiological profiles. Appl. Environ. Microbiol. 65: 982-988
Felsenstein, J. 1995. PHYLIP package version 3.5. Department of Genetics, University of Washington, Seattle, WA
Games, L. M., K. L. Dickson, A. W. Maki and J. Cairns. 1982. Field valication of exposure analysis modeling system (EXAMS) in “Modeling the fate of chemicals in the aquatic environment”. Ann Arbor Science, pp 325-346
Huang, Y.-F and S.-L. Huang. 2001. Personal communications.
Hummerjohann, J., E. Kuttel, M. Quadroni, J. Ragaller, T. Leisinger and M. A. Kertesz. 1998. Regulation of the sulfate starvation response to Pseudomonas aeruginosa: role of cysteine biosynthetic intermediates. Microbiology. 141: 1375-1386
Humphrey, R. E., M. H. Ward and W. Hinze. 1970. Spectrophotometric determination of sulfite with 4,4’-dithiodipyridine and 5,5’-dithiobis-(2-nitrobenzoic acid). Anal. Chem. 42: 698-702
Jimenez, L., A. Breen, N. Thomas, T. W. Federle, and G. S. Sayler. 1991. Mineralization of linear alkylbenzene sulfonate by a four-member aerobic bacterial consortium. Appl. Environ. Microbiol. 57: 1566-1569
Johnston, J. B., K. Murray and R. B. Cain. 1975. Microbial metabolism of aryl sulphonates. A reassessment of colorimetric methods for the determination of sulphite and their use in measuring desulphonation of aryl and alkylbenzene sulphonates. Antonie van Leeuwenhoek 41: 493-511
Junker, F., T. Leisinger and A. M. Cook. 1994. 3-Sulphocatechol 2,3-dioxygenase and other dioxygenases (EC 1.13.11.2 and EC 1.14.12.-) in the degradative pathways of 2-aminobenzenesulphonic, benzenesulphonic and 4-toluenesulphonic acids in Alcaligenes sp. strain O-1. Microbiology. 140: 1713-1722
Junker, F. and A. M. Cook. 1997. Conjugative plasmids and the degradation of arylsulfonates in Commamonas testosteroni. Appl. Environ. Microbiol. 63: 2403-2410
Junker, F., R. Kiewitz and A. M. Cook. 1997. Characterization of the p-toluenesulfonate operon tsaMBCD and tsaR in Commamonas testosteroni T-2. J. Bacteriol. 179: 919-927
Kahnert, A., P. Vermeij, C. Wietek, P. James, T. Leisinger and M. A. Kertesz. 2000. The ssu locus plays a key role in organosulfur metabolism in Pseudomonas putida S-313. J. Bacteriol. 182: 2869-2878
Kertesz, M. A. 1999. Riding the sulfur cycle — metabolism of sulfonates and sulfate esters in Gram-negative bacteria. FEMS Microbiol. Rev. 24: 135-175
Kertesz, M. A., P. Kolbener, H. Stockinger, S. Beil, and A. M. Cook. 1994. Desulfonation of linear alkylbenzenesulfonate surfactants and related compounds by bacteria. Appl. Environ. Microbiol. 60: 2296-2303
Laemmli, U. K. 1970. Cleavage of structure proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680
Lajoie, C. A., A. C. Layton and G. S. Sayler. 1994. Cometabolite oxidation of polychlorinated biphenyls in soil with a surfactant-based field application vector. Appl. Environ. Microbiol. 60: 2826-2833
Locher, H. H., T. Leisinger and A. M. Cook. 1991. 4-Sulphobenzoate 3,4-dioxygenase: purification and properties of a desulphonative two-component enzyme system from Comamonas testosteroni T-2. Biochem. J. 274: 833-842
MicrologTM system, Release 4.0 User Guide. 1998. Section 13. pp.3-20.
Nielsen, A. T., W. Liu, C. Filipe, L. Grady, S. Molin and D. 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
Olsen, G. J., C. R. Woese and R. Overbeek. 1994. The winds of (evolutionary) changes: breathing new life into microbiology. J. Bacteriol. 178: 1-6
Prescott, L. M., J. P. Harley and D. A. Klein. 1999. Microbiology, 4th ed., WCB/McGraw-Hill
Quadroni, M., W. Staudenmann, M. A. Kertesz and P. James. 1996. Analysis of global responses by protein and peptide fingerprinting of proteins isolated by two-dimensional gel electrophoresis: application to the sulfate-starvation response of Escherichia coli. Eur. J. Biochem. 239: 773-781
Raoult, D. Submitted (10-JUN-1999) Unite des Rickettsies, Faculte de Medecine, 27, Boulevard Jean Moulin, Marseille 13385, France
Reichenbecher, W. and J. C. Murrell. 1999. Linear alkanesulfonates as carbon and energy sources for gram-positive and gram-negative bacteria. Arch. Microbiol. 171: 430-438
Reiff, B., R. Lloyd, M. J. How, D. Brown and J. S. Alabaster. 1979.
The acute toxicity of eleven detergents to fish: Results of an interlaboratory exercise. Wat. Res. 13: 207-210
Richard, C. 1974. A new species of enterobacteria found in medical bacteriology: Enterobacter agglomerans (ex-Erwinia herbicola). Biochemical study of 205 strains. Ann. Biol. Clin. 32: 341-346
Sigoillot, J-C. and M.H. Nguyen. 1992. Complete oxidation of linear alkylbenzene sulfonate by bacterial communities selected from coastal seawater. Appl. Environ. Microbiol. 58: 1308-1312
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. Cohen-Bazire and W. R. Sistrom. 1957. Kinetics studies of pigment synthesis by non-sulfur purple bacteria. J. Cell. Comp. Physiol. 49:25.
Swisher, R. D. 1987. Surfactant biodegradation, 2nd ed. Marcel Dekker, New York.
Vermeij, P., C. Wietek, A. Kahnert, T. Wuest and M. A. Kertesz. 1999. Genetic organization of sulfur-controlled aryl desulfonation in Pseudomonas putida S-313. Mol. Microbiol. 32: 913-926
Vermeij, P. and M. A. Kertesz. 1999. Pathways of assimilative sulfur metabolism in Pseudomonas putida. J. Bacteriol. 181: 5833-5837
White, G. F. and N. J. Russell. 1993. Biodegradation of anionic surfactants and related molecules. In: Ratledge C (Ed) Biochemistry of Microbial Degradation. Kluwer Academic Press Dordrecht.
Willetts, A. J. and R. B. Cain. 1972. Microbial metabolism of alkylbenzene sulfonates. Bacterial metabolism of undecylbenzene-p-sulphonate and dodecylbenzene-p-sulfonate. Biochem. J. 129: 389-402
Zurrer, D., A. M. Cook and T. Leisinger. 1987. Microbial desulfonation of substituted naphthalenesulfonic acids and benzenesulfonic acids.
Appl. Environ. Microbiol. 53: 1459-1463
指導教授 黃雪莉(Shir-Ly Huang) 審核日期 2001-7-20
推文 facebook   plurk   twitter   funp   google   live   udn   HD   myshare   reddit   netvibes   friend   youpush   delicious   baidu   
網路書籤 Google bookmarks   del.icio.us   hemidemi   myshare   

若有論文相關問題,請聯絡國立中央大學圖書館推廣服務組 TEL:(03)422-7151轉57407,或E-mail聯絡  - 隱私權政策聲明