跳到主要內容

臺灣博碩士論文加值系統

(3.236.110.106) 您好!臺灣時間:2021/07/29 17:32
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:林哲毅
研究生(外文):Chei-Yi Lin
論文名稱:炭角菌之多氯聯苯分解酵素的純化與探討
論文名稱(外文):Isolation, purification and characterization of the PCBs-degrading enzyme from Xylaria regalis
指導教授:張勝祺張勝祺引用關係
指導教授(外文):Shenq-Chyi Chang
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物化學研究所
學門:生命科學學門
學類:生物化學學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:炭角菌多氯聯苯
外文關鍵詞:Xylaria regalisPCB
相關次數:
  • 被引用被引用:1
  • 點閱點閱:134
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
多氯聯苯(Polychlorinated Biphenyls,簡稱PCBs)是一種廣泛存在於環境之中的環境污染物,許多研究更已証實其對於人類和動物會造成危害。本實驗室發現木生性腐生真菌Xylaria regalis對於多氯聯苯具有分解能力,根據本實驗室之前的研究,Xylaria regalis在以SDB培養基進行培養的條件之下,其分解多氯聯苯的能力最佳,因此本實驗室便以SDB培養基來培養Xylaria regalis,並進一步純化其多氯聯苯分解酵素。
本實驗室將多氯聯苯添加到土壤中,仿照環境污染的情況,以SDB菌株培養液探討其分解土壤中多氯聯苯之情形。結果顯示,第十天的SDB菌株培養液在30 ℃下,反應2小時,能分解67 %的Aroclor 1254 (PCBs)。進一步以陰離子交換樹脂(QAE Sephadex A-25)和疏水性層析管柱(Phenyl-650M)純化其多氯聯苯分解酵素,所得到的酵素純度提高到11.60倍,比活性為8.90 x 10-2 unit/mg,回收率為108.29 %。
根據高效率液相層析儀之膠體過濾層析管柱和SDS-PAGE,得知酵素分子量約為22.5 kDa。酵素最佳反應pH值為7,最佳反應溫度在30 ℃,而且在溫度30 ℃及pH 7的環境下,酵素最為穩定。本實驗室所研究的金屬離子中,鐵離子對酵素活性有明顯的抑制效果;EDTA對酵素活性同樣也有抑制效果。
PCBs (Polychlorinated Biphenyls) are widespread pollutants. Many studies have proven that PCBs are hazardous to humans and other animals. Xylaria regalis is a wood-grown saprophytic fungus. Based on previous study, Xylaria regalis is capable of PCBs degradation, and the fungus could degrade more PCBs in SDB medium than in other media. So, We cultured Xylaria regalis in SDB medium and further purified the PCBs-degrading enzyme.
We found that culture medium from 10-day growth mycelia could degrade 67 % of Aroclor 1254 (PCBs). And the PCBs-degrading enzyme was further purified by anion exchanger column (QAE Sephadex A-25) and hydrophobic interaction column (Phenyl-650M). The purity of the PCBs-degrading enzyme was increased to 11.60 fold, the specific activity of the PCBs-degrading enzyme was reached to 8.90 x 10-2 unit/mg, and the recovery of the purified enzyme was 108.29 %.
The molecular weight of the PCBs-degrading enzyme was found to be approximately 22.5 kDa by HPLC gel filtration and SDS-PAGE. The optimal pH of the enzyme was 7 and the optimal temperature of the enzyme was 30 ℃. The pH stability of the enzyme was 7 and the thermostability of the enzyme was 30 ℃. The iron ions that we studied in different concentration can inhibit the enzyme activity. When EDTA was added in the enzyme solution, the enzyme activity was inhibited.
1.Boyle A.W., Silvin C.J., Hassett J.P., Nakas J.P., and Tanenbaum S.W. (1992) Bacterial PCB biodegradation. Biodegradation 3, 285 ~ 298.
2.Chang K.J., Chen J.S., and Huang P.C. (1980) Studies of patients with polychlorinated biphenyls poisoning. Blood analysis of patients. J. Formosa Med. Assoc. 79, 304 ~ 313.
3.Chao H.R., Wang S.L., Lin L.Y., Yu H.Y., Lu Y.K., Chou W.L., Guo Y.L., and Chang L.W. (2003) Polychlorinated biphenyls in Taiwanese primipara human milk and associated factors. Bull. Environ. Contam. Toxicol. 70, 1097 ~ 1103.
4.Chao H.R., Wang S.L., Lee C.C., Yu H.Y., Lu Y.K., and Papke O. (2004) Level of polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls (PCDD/Fs, PCBs) in human milk and the input to infant body burden. Food Chem Toxicol. 42, 1299 ~ 1308.
5.Chen P.H. (1981) Polychlorinated biphenyls: environmental occurrence, biological and toxicological effects. Cli. Med. (Taipei) 7, 4 ~ 8.
6.Chen P.H., Chang K.T., and Lu Y.D. (1981) Toxic compounds in the cooking oil which caused PCB poisoning in Taiwan. Levels of polychlorinated biphenyls and polychlorinated dibenzofurans. Cli. Med. (Taipei) PCB Poisoning Special Issue. 7, 71 ~ 76.
7.Chen P.H., Lu Y.D., and Yang M.H. (1981) Toxic compounds in the cooking oil which caused PCB poisoning. The presence of polychlorinated quaterphenyls and polychlorinated terphenyls. Cli. Med. (Taipei). PCB Poisoning Special Issue. 7, 77 ~ 82.
8.Gladen B.C., Schecter A.J., Papke O., Shkyryak-Nyzhnyk Z.A., Hryhorczuk D.O., and Little R.E. (1999) Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and coplanar polychlorinated biphenyls in breast milk from two cities in Ukraine. J Toxicol Environ Health A 58, 119 ~ 127.
9.Guo Y.L., Ryan B.P.Y., Lau B.Y.P., Yu M.L., and Hsu C.C. (1997) Blood serum levels of PCBs and PCDFs in Yucheng women 14 years after exposure to a toxic rice oil. Archives of Environmental Contamination and Toxicology. 33, 104 ~ 108.
10.Jensen S. (1966) A new chemical hazard. New Sci. 32, 612.
11.Karlson K., Ishaq R., Becker G., Berggren P., Broman D., and Colmsjo A. (2000) PCBs, DDTs and methyl sulphone metabolites in various tissues of harbour porpoises from Swedish waters. Environ Pollut. 110, 29 ~ 46.
12.Kimbrough R.D., and Krouskas C.A. (2003) Human exposure to polychlorinated biphenyls and health effects : a critical synopsis. Toxicol Rev. 22, 217 ~ 233.
13.Kuratsune M., Yoshimura T., and Matsuzaka J. (1972) Yusho, a poisoning caused by rice oil contaminated with chlorobiphenyls. Environ. Health. Perspec. 1, 119 ~ 128.
14.Lopez-Avila V., Benedicto J., Charan C., and Young R. (1995) Determination of PCBs in soil/sediments by microwave-assisted extraction and GC/ECD or ElISA. Environ. Sci. Technol. 29, 2709 ~ 2712.
15.Mayes B.A., McConnell E.E., Neal B.H., Brunner M.J., Hamilton S.B., Sullivan T.M., Peters A.C., Ryan M.J., Toft J.D., Singer A.W., Brown J.F.Jr., Menton R.G., and Moore J.A. (1998) Comparative carcinogenicity in Sprague-Dawley rats of the polychlorinated biphenyl mixtures Aroclors 1016, 1242, 1254, and 1260. Toxicol. Sci. 41, 62 ~ 76.
16.Oakley G.G., Devanaboyina U., Robertson L.W., and Gupta R.C. (1996) Oxidative DNA damage induced by activation of polychlorinated biphenyls (PCBs): implications for PCB-induced oxidative stress in breast cancer. Chem. Res. Toxicol. 9, 1285 ~1292.
17.Pineiro M.E.A., Lozano J.S., and Yusty M.A.L. (1996) Comparison of two extraction methods for determination of PCBs and PCTs in mussels from Galicia. Talanta 43, 487 ~ 491.
18.Robinson G.K., and Lenn M.J. (1994) The bioremediation of polychlorinated biphenyls (PCBs): problems and perspectives. Biotechnology and Genetic Engineering Reviews 12, 139 ~ 188.
19.Rossi L., de Alencastro L., Kupper T., and Tarradellas J. (2004) Urban stormwater contamination by polychlorinated biphenyls (PCBs) and its importance for urban water systems in Switzerland. Sci Total Environ. 322, 179 ~ 189.
20.Saito R., Shigematsu N., and Ishimam S. (1972) Immunoglobulin levels in serum and sputum of patients with PCB poisoning. Fukuoka Acta. Med. 63, 408 ~ 411.
21.Schuhmacher M., Nadal M., and Domingo J.L. (2004) Levels of PCDD/Fs, PCBs, and PCNs in soils and vegetation in an area with chemical and petrochemical industries. Environ Sci Technol. 38, 1960 ~ 1969.
22.Silberhorn E.M., Glauert H.P., and Robertson L.W. (1990) Carcinogenicity of polyhalogenated biphenyls: PCBs and PBB. Crit. Rev. Toxicol. 20, 439 ~ 496.
23.Smedes F., and de Boer J. (1997) Determination of chlorobiphenyls in sediments---analytical methods. Trends in Anal. Chem. 16, 503 ~ 517.
24.Urabe H., Koda H., and Asahi M. (1979) Present state of Yusho patients. Ann. N. Y. Acad. Sci. 320, 263 ~ 276.
25.Uzawa H., Notomi A., and Nakamuta S. (1972) Consecutive three year follow up study of serum triglyceride concentrations of 82 subjects with PCB poisoning. Fukuoka Acta. Med. 63, 401 ~ 404.
26.Vit L. (1992) Polychlorinated biphenyls in the environment. J. Chrom. 595, 1 ~ 43.
27.Yamaguchi A., Yoshimura T., and Kuratsune M. (1971) A survey on pregnant women having consumed rice oil contaminated with chlorobiphenyls and their babies. Fukuoka Acta. Med. 62, 117 ~ 122.
28.Yoshimura H., Yamamoto H., and Uzawa H. (1971) Studies on the tissue distribution and the urinary and facal excretion of 3H-Kanechlor (chlorobiphenyls) in rats. Fukuoka Acta. Med. 62, 12 ~ 19.
29.Yoshimura T. (1974) Epidemiological study on Yusho babies born to mothers who had consumed oil contaminated by PCB. Ibid. 65, 74 ~ 80.
30.Zuloaga O., Etxebarria N., Fernández L.A., and Madariaga J.M. (1998) Comparison of accelerated solvent extraction with microwave-assisted extraction and Soxhlet for the extraction of chlorinated biphenyls in soil samples. Trends in Anal. Chem. 17, 642 ~ 647.
31.高銘木(1990),環境保護---多氯聯苯對生態環境的毒害,工業污染防治,36,1 ~ 7。
32.饒連財(1980),多氯聯苯(PCBs)與米糠油中毒事件,環境科學,1, 33 ~ 49。
33.饒連財(1985),多氯聯苯(PCBs)之毒理學---台灣多氯聯苯事件的探討與污染,污染毒理學,24 ~ 65。
34.行政院(2002),環署檢字第0910014627號公告:去硫淨化法。
35.行政院(2002),環署檢字第0910077075號公告:土壤及事業廢棄物中多氯聯苯檢測方法---毛細管柱氣相層析法。
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top