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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39236完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃穰(Rang Huang),周宏農(Hong-Nong Chou) | |
| dc.contributor.author | Yi-Min Chen | en |
| dc.contributor.author | 陳逸民 | zh_TW |
| dc.date.accessioned | 2021-06-13T17:24:39Z | - |
| dc.date.available | 2005-01-27 | |
| dc.date.copyright | 2005-01-27 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-01-26 | |
| dc.identifier.citation | Anderson R. J., Luu H. A., Chen D. X. X., Holmes C. F. B., Kent M. L., Blanc M. L., ‘Max’ Taylor F. J. R. and Williams D. E., 1993. Chemical and biological evidence links microcystins to salmon ‘Netpen liver disease’. Toxicon, 31: 1315-1323.
An, J. and Carmichael, W. W., 1994. Use of a colorimetric protein phosphatase inhibition assay and enzyme linked immunosorbent assay for the study of microcystins and nodularins. Toxicon, 32: 1495-1507. Ash C., Mackintosh C, Mackintosh R. and Fricker C. R., 1995. Use of a protein phosphatase inhibition test for the detection of cyanobacterial toxins in water. Wat. Sci. Tech., 31: 51-53. Aune T. and Berg K., 1986. Use of freshly prepared rat hepatocytes to study toxicity of blooms of the blue-green algae Microcystis aeruginosa and Oscillatoria agardhii. J. Toxicol. Environ. Health., 19: 325-336. Bagu J. R., Sykes B. D., Craig M. M. and Holmes C. F. B., 1997. A molecular basis for different interactions of marine toxins with protein phosphatase-1. J. Biol. Chem., 272: 5087-5097. Bajuk A., Gluch K. and Michalak L., 2001. Effect of impurities on the matrix-assisted laser desorption/ionization mass spectra of insulin. Rapid Commun. Mass Spectrom., 15: 2383-2386. Barford D., 1996. Molecular mechanisms of the protein serine/threonine phosphatases. Trends Biochem. Sci., 21: 407-412. Bialojan C. and Takai A., 1988. Inhibitory effect of a marine-sponge toxin, okadaic acid, on protein phosphatases. Biochem. J., 256: 283-290. Blom J. F., Robinson J. A. and Juttner F., 2001. Higher grazer toxicity of [D-Asp3, (E)-Dhb7] microcystin-RR of Planktothrix rubescens as compared to different microcystins. Toxicon, 39: 1923-1932. Bornsen K. O., Gass M. A. S., Bruin, G. J. M., von Adrichem, J. H. M., Biro, M. C., Kresbach, G. M. and Ehrat, M., 1997. Influence of solvents and detergents on matrix-assisted laser desorption/ionization mass spectrometry measurements of proteins and oligonucleotides. Rapid Commun. Mass. Spectrom., 11: 603-609. Botes D. P., Kruger H. and Viljoen C. C., 1982. Isolation and characterization of four toxins from the blue-green alga, Microcystis aeruginosa. Toxicon, 20: 945-954. Bury, N. R., Codd, G. A., Wendelaar Bonga S. E. and Flik, G., 1998. Fatty acids from the cyanobacterium Microcystis aeruginosa with potent inhibitory effects on fish gill Na+/K+-ATPase activity. J. Exp. Biol., 201: 81-89. Campbell D. L., Lawton L. A., Beattle K. A. and Codd G. A., 1994. Comparative assessment of the specificity of the brine shrimp and microtox assays to hepatotoxic (microcystin-LR-containing) cyanobacteria. Environ. Toxicol. Wat. Qual., 9: 71-77. Carbis C. R., Mitchell G. F., Anderson J. W. and McCauley I., 1996. The effects of microcystins on the serum biochemistry of carp, Cyprinus carpio L., when the toxins are administered by gavage, immension and intraperitoneal routes. J. Fish Dis., 19: 151-159. Carmichael W. W., 1988. Freshwater cyanobacteria (blue-green algae) toxins. In: Natural toxins. Ownby C. L. and Odell G.. V. (Eds), Pergamon Press, New York, pp. 3-16. Carmichael W. W., Beasely V., Bunner D. L., Eloff J. N., Falconer I., Gorham P., Harada K. I., Krishnamurthy T., Yu M. J., Moore R. E., Rinehart K., Runnegar M., Skulberg O. M. and Watanabe M., 1988. Naming of cyclic heptapeptide toxins of cyanobacteria (blue-green algae). Toxicon, 26: 971-973. Carpenter E. J. and Carmichael W. W., 1995. Taxonomy of cyanobacteria. In: Manual on Harmful Marine Microalgae. Hallegraeff G. M., Anderson D. M., Cembella A. D. and Enevoldsen H. O. (Eds). UNESCO, France, pp. 373-380. Cembella A. D., Milenkovic L. V. and Doucette G. J., 1995. Part A. In Vitro biochemical and cellular assays. In: Manual on Harmful Marine Microalgae. Hallegraeff G. M., Anderson D. M., Cembella A. D. and Enevoldsen H. O. (Eds), UNESCO, France, pp.181-212. Chen Y. M., Lee T. H., Huang R., Bai J. Y. and Chou, H. N., 2004. Comparison of protein phosphatase inhibition assay with the mouse and Artemia toxicity assays in the toxic Microcystis screening. Aquat. Toxicol., in submission. Chong M. W. K., Gu K. D., Lam P. K. S., Yang M. and Fong W. F., 2000. Study on the cytotoxicity of microcystin-LR on cultured cells. Chemosphere, 41: 143-147. Claffey D. J. and Ruth J. A., 2001. Amphetamine adducts of melanin intermediates demonstrated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Chem. Res. Toxicol., 14: 1339-1344. Cohen P., 1989. The structure and regulation of protein phosphatase. Annu. Rev. Biochem., 58: 453-508. Cohen P., Alemany S., Hemmings B.A., Resink T.J., Stralfors P. and Tung H.Y., 1988. Methods Enzymol. 159, 390-408. Connor J. H., Kleeman T., Barik S., Honkanen R. E. and Shenolikar S., 1999. Important of the b12-b13 loop in protein phosphatase-1 catalytic subunit for inhibition by toxins and mammalian protein inhibitors. J. Biol. Chem., 274: 22366-22372. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39236 | - |
| dc.description.abstract | 首先以現有的微囊藻株為材料，分別測試其對於小白鼠、豐年蝦的毒性，以及其對於蛋白質磷酸水解酵素PP-1c的抑制活性後，分析此三種活性表現之關連性。由對於PP-1c具有抑制活性的藻株，亦對兩種生物皆具明顯毒性的初步結果，推測微囊藻毒對於兩種生物的毒性，可能均與其抑制蛋白質磷酸水解酵素的活性有關。在以自行設計的矽膠固相萃取法處理粗萃樣品後，將使三種活性分析數值間一致性更佳的結果，說明本法的使用，有助於改善上述分析方法對於微囊藻毒分析的精確性。其對於微囊藻樣品內各種微囊藻毒的回收率，另經後續MALDI-TOF及HPLC的分析而獲肯定。
後續以由上述藻株中所純化出之各種微囊藻毒純質，分別測定其對於小白鼠的急性毒性、PP-1c及PP-2Ac兩種蛋白質磷酸水解酵素的抑制活性，以及其與PP-1c形成共價產物之動力學。藉由微囊藻毒結構與活性關連性數值的分析，推測出微囊藻毒的第三氨基酸MeAsp，在抑制PP-2Ac的活性表現中所扮演的角色，並不若其於抑制PP-1c來的重要，同時得知微囊藻毒的急性毒性和PP-2Ac抑制的關連，遠高於其對PP-1c的抑制。在以MALDI-TOF分析MCYST-LR與PP-1c共價鍵形成動力學的研究中，得知在30oC的反應條件下，每小時僅有低於5 %的PP-1c會形成共價鍵，反應速率極為緩慢。此說明共價鍵的形成應為毒素以其他非共價鍵的分子間作用力與PP-1c結合，同時抑制其活性後再出現的事件，與毒素的抑制活性間並無直接的關連。 | zh_TW |
| dc.description.abstract | Strains of Microcystis have been cultured for their toxicity analysis, and their toxins were isolated for their specific toxicity tests. Mouse and Artemia toxicity assays were applied for the various strains and their respective LD50 and LC50 that were compared with the IC50 of protein phosphatase inhibition assay after converted to the toxin content of MCYST-LR equivalent. Strains that do not possess any PP1 inhibitory activity show no toxicity to either mice or brine shrimp larvae. Ranking of the animal toxicities coincided well with enzyme inhibitory activities. A solid phase extraction procedure was developed for the removal of interfering agents in Artemia assay and proved to be very effective. This SPE pretreatment in conjugation with the following assays can be applied for the MCYST screening in field cyanobacterial samples or in algal food supplements. MALDI-TOF and HPLC metabolomic analysis showed the SPE good in preserving the toxins and removing contaminants.
In the further study a series of microcystins were tested on the mouse toxicity, protein phosphatase inhibition activities of PP-1 and PP-2A from native or recombinant sources, and the covalent-bonding kinetics with recombinant PP-1c. Structure-Activity-Relationship studies showed mouse toxicities of different microcystins were in better correlation with their inhibitory activities against PP2Ac than PP1c. Different microcystins have different inhibitory activities on different enzyme sources, indicating the different interaction in molecular level. The covalent-bonding kinetics obtained from MALDI-TOF assays showed that less than 5% of total PP-1c covalent-bonding with MCYST-LR per hour under 30oC, revealing the slow reacting nature of covalent-bonding formation between PP-1c and microcystin-LR. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T17:24:39Z (GMT). No. of bitstreams: 1 ntu-94-D88241003-1.pdf: 6032705 bytes, checksum: 53300c2c703f36eb319d879eed3d9ce4 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 摘要 ---------------------------------------------------------------------------- iii
壹、文獻回顧 ----------------------------------------------------------------- 1 貳、前言 ---------------------------------------------------------------------- 29 參、研究一、微囊藻株的生物毒性與蛋白質磷酸水解酵素的活性抑制分析 一、摘要 ------------------------------------------------------------------------------- 37 二、材料方法 ------------------------------------------------------------------------- 38 三、結果 ------------------------------------------------------------------------------- 46 四、討論 ------------------------------------------------------------------------------- 55 肆、研究二、微藻健康食品的微囊藻毒高效液相層析與蛋白質磷酸水解酵素活性抑制分析 一、摘要 ------------------------------------------------------------------------------- 59 二、材料方法 ------------------------------------------------------------------------- 60 三、結果 ------------------------------------------------------------------------------- 64 四、討論 ------------------------------------------------------------------------------- 70 伍、研究三、應用介質輔助激光脫附離子化-飛行時間質譜儀於微囊藻毒素組成的分析 一、摘要 ------------------------------------------------------------------------------- 73 二、材料方法 ------------------------------------------------------------------------- 74 三、結果 ------------------------------------------------------------------------------- 76 四、討論 ------------------------------------------------------------------------------- 81 陸、研究四、微囊藻毒化學結構與其蛋白質磷酸水解酵素活性抑制之關連性研究 一、摘要 ------------------------------------------------------------------------------- 85 二、材料方法 ------------------------------------------------------------------------- 87 三、結果 ------------------------------------------------------------------------------- 88 四、討論 ------------------------------------------------------------------------------- 96 柒、研究五、微囊藻毒MCYST-LR與蛋白質磷酸水解酵素PP-1c共價產物形成的動力學研究 一、摘要 ------------------------------------------------------------------------------- 99 二、材料方法 ------------------------------------------------------------------------ 100 三、結果 ------------------------------------------------------------------------------104 四、討論 ------------------------------------------------------------------------------113 捌、結論 ---------------------------------------------------------------------- 119 玖、參考文獻 --------------------------------------------------------------- 121 拾、附錄 一、論文中常用英文縮寫與全名之對照表 ----------------------------------- 145 二、修正 Fitzgerald 培養基配方 -----------------------------------------------147 三、個人著述與發表 ---------------------------------------------------------------149 | |
| dc.language.iso | zh-TW | |
| dc.subject | 微囊藻毒 | zh_TW |
| dc.subject | microcystin | en |
| dc.title | 微囊藻毒檢測方法與毒理機制的探究 | zh_TW |
| dc.title | Studies on the Detection and Toxicology of Microcystins | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-1 | |
| dc.description.degree | 博士 | |
| dc.contributor.advisor-orcid | ,周宏農(unijohn@ntu.edu.tw) | |
| dc.contributor.oralexamcommittee | 宋賢一(Hein-Yi Sung),余豐益(Feng-Yi Yu),黃憲斌(Hsin-Bin Huang),吳俊宗(Jun-Tzong Wu) | |
| dc.subject.keyword | 微囊藻毒, | zh_TW |
| dc.subject.keyword | microcystin, | en |
| dc.relation.page | 146 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-01-26 | |
| dc.contributor.author-college | 理學院 | zh_TW |
| dc.contributor.author-dept | 海洋研究所 | zh_TW |
| 顯示於系所單位： | 海洋研究所 | |
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