Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/3263
DC FieldValueLanguage
dc.contributor劉永銓zh_TW
dc.contributor.author連瑋翎zh_TW
dc.contributor.authorLien, Wei-Lingen_US
dc.contributor.other化學工程學系所zh_TW
dc.date2012en_US
dc.date.accessioned2014-06-06T05:31:35Z-
dc.date.available2014-06-06T05:31:35Z-
dc.identifierU0005-1508201215475000en_US
dc.identifier.citation1. 陳光宇, 大腸桿菌醱酵生產盤尼西林 G 醯胺酵素代謝工程之研究. 2009. 2. 黃宏彰, 應用基因重組大腸桿菌生產盤尼西林醯胺酵素之醱酵條件探討. 2001. 3. Sulkowski, E., Purification of proteins by IMAC. Trends in biotechnology, 1985. 3(1): p. 1-7. 4. Hemdan, E.S. and J. Porath, Development of immobilized metal affinity chromatography: II. Interaction of amino acids with immobilized nickel iminodiacetate. Journal of Chromatography A, 1985. 323(2): p. 255-264. 5. Wei, Q.B.S. and R. Yao, Progress of Immobilized Metal-chelate Affinity Chromatography. 6. Zou, H., Q. Luo, and D. Zhou, Affinity membrane chromatography for the analysis and purification of proteins. Journal of Biochemical and Biophysical Methods, 2001. 49(1): p. 199-240. 7. 何立凡, 金屬親和吸附材於蛋白質純化及酵素固定化之應用. 2001. 8. Lihme, A., et al., Divinylsulphone-activated agarose:: Formation of stable and non-leaking affinity matrices by immobilization of immunoglobulins and other proteins. Journal of Chromatography B: Biomedical Sciences and Applications, 1986. 376: p. 299-305. 9. 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Morganti, Current and prospective applications of metal ion–protein binding. Journal of Chromatography A, 2003. 988(1): p. 1-23. 15. Chaga, G.S., Twenty-five years of immobilized metal ion affinity chromatography: past, present and future. Journal of Biochemical and Biophysical Methods, 2001. 49(1-3): p. 313. 16. Jr-Chiang, J.J., 應用金屬親和薄膜分離純化盤尼西林醯胺酵素. 2002. 17. Chaga, G.S., B. Ersson, and J.O. Porath, Isolation of calcium-binding proteins on selective adsorbents application to purification of bovine calmodulin. Journal of Chromatography A, 1996. 732(2): p. 261-269. 18. Porath, J., B. Olin, and B. Granstrand, Immobilized-metal affinity chromatography of serum proteins on gel-immobilized group III A metal ions. Arch Biochem Biophys, 1983. 225(2): p. 543-547. 19. Birger Anspach, F., Silica-based metal chelate affinity sorbents I. Preparation and characterization of iminodiacetic acid affinity sorbents prepared via different immobilization techniques. Journal of Chromatography A, 1994. 672(1-2): p. 35-49. 20. Liu, Y.C., et al., Effects of spacer arm on penicillin G acylase purification using immobilized metal affinity membranes. Journal of membrane science, 2005. 251(1): p. 201-207. 21. Todd, R.J., R.D. Johnson, and F.H. Arnold, Multiple-site binding interactions in metal-affinity chromatography:: I. Equilibrium binding of engineered histidine-containing cytochromes c. Journal of Chromatography A, 1994. 662(1): p. 13-26. 22. Johnson, R.D., R.J. Todd, and F.H. Arnold, Multipoint binding in metal-affinity chromatography II. Effect of pH and imidazole on chromatographic retention of engineered histidine-containing cytochromes< i> c</i>. Journal of Chromatography A, 1996. 725(2): p. 225-235. 23. Johnson, R.D. and F.H. Arnold, Review: Multipoint binding and heterogeneity in immobilized metal affinity chromatography. Biotechnology and Bioengineering, 1995. 48(5): p. 437-443. 24. Finette, G., Q.M. Mao, and M.T.W. Hearn, Comparative studies on the isothermal characteristics of proteins adsorbed under batch equilibrium conditions to ion-exchange, immobilised metal ion affinity and dye affinity matrices with different ionic strength and temperature conditions. Journal of Chromatography A, 1997. 763(1): p. 71-90. 25. Porath, J., et al., Metal chelate affinity chromatography, a new approach to protein fractionation. Nature, 1975. 258: p. 598-599. 26. Porath, J., IMAC—Immobilized metal ion affinity based chromatography. TrAC Trends in Analytical Chemistry, 1988. 7(7): p. 254-259. 27. Hubert, P. and J. Porath, Metal chelate affinity chromatography:: I. Influence of various parameters on the retention of nucleotides and related compounds. Journal of Chromatography A, 1980. 198(3): p. 247-255. 28. Gibert, S., N. Bakalara, and X. Santarelli, Three-step chromatographic purification procedure for the production of a His-tag recombinant kinesin overexpressed in E. coli. Journal of Chromatography B: Biomedical Sciences and Applications, 2000. 737(1-2): p. 143-150. 29. Fitton, V. and X. Santarelli, Evaluation of immobilized metal affinity chromatography for purification of penicillin acylase. Journal of Chromatography B: Biomedical Sciences and Applications, 2001. 754(1): p. 135-140. 30. Hu, H.L., et al., Purification of VP3 protein of infectious bursal disease virus using nickel ion-immobilized regenerated cellulose-based membranes. Journal of Chromatography B, 2006. 840(2): p. 76-84. 31. Suen, S.Y., Y.C. Liu, and C.S. Chang, Exploiting immobilized metal affinity membranes for the isolation or purification of therapeutically relevant species. Journal of Chromatography B, 2003. 797(1-2): p. 305-319. 32. Wu, C.Y., et al., Analysis of protein adsorption on regenerated cellulose-based immobilized copper ion affinity membranes. Journal of Chromatography A, 2003. 996(1): p. 53-70. 33. Liu, Y.C., C.C. ChangChien, and S.Y. Suen, Purification of penicillin G acylase using immobilized metal affinity membranes. Journal of Chromatography B, 2003. 794(1): p. 67-76. 34. 潘建亮, 固定化盤尼西林去醯基酵素反應動力學 建模及其兩水相系統分離反應之探討. 成功大學化學工程學系碩博士班學位論文, 2005(2005 年). 35. Yocum, R.R., J. Rasmussen, and J. Strominger, The mechanism of action of penicillin. Penicillin acylates the active site of Bacillus stearothermophilus D-alanine carboxypeptidase. Journal of Biological Chemistry, 1980. 255(9): p. 3977-3986. 36. 陳國誠, 生物固定化技術與產業應用. 茂昌圖書有限公司, Taiwan, 2000. 37. 洪佃玠, 利用幾丁聚醣固定脂肪分解酵素之研究. 2002. 38. CHONG AI SHING, M., Immobilization of enzyme penicillin G acylase on functionalized nanoporous silicas for biocatalysis applications. 2004. 39. Dixon, M., et al., Enzymes. 1979, New York: Academic Press. 40. 呂鋒洲 and 林仁混, 基礎酵素學. 1994. 41. Jochems, P., et al., Enzyme immobilization on/in polymeric membranes: status, challenges and perspectives in biocatalytic membrane reactors (BMRs). Green Chemistry, 2011. 13(7): p. 1609-1623. 42. Parmar, A., et al., Advances in enzymatic transformation of penicillins to 6-aminopenicillanic acid (6-APA). Biotechnology advances, 2000. 18(4): p. 289-301. 43. Kheirolomoom, A., et al., Clarification of penicillin G acylase reaction mechanism. Process Biochemistry, 2001. 36(11): p. 1095-1101. 44. Žuža, M., B. Obradović, and Z. Knežević‐Jugović, Hydrolysis of Penicillin G by Penicillin G Acylase Immobilized on Chitosan Microbeads in Different Reactor Systems. Chemical Engineering & Technology, 2011. 34(10): p. 1706-1714.en_US
dc.identifier.urihttp://hdl.handle.net/11455/3263-
dc.description.abstract盤尼西林醯胺酵素(penicillin G acylase, PGA)是一重要生化觸媒,使盤尼西林G(penicillin G, PG)進行水解反應生成六青黴素酸(6-aminopenicillanic acid, 6-APA),它和一些有機物以有機合成反應形成β-lactam抗生素。在全球巿場,每年有數千噸β-lactam抗生素需求量應用於治療方面。因此製備出高活性與穩定性之PGA對於6-APA生產是很重要。 本研究以再生纖維膜(regenerated cellulose-based membrane)作為載體,先使用EPI (epichlorohydrin),再利用化學方法接上IDA(iminodiacetic acid)螯合劑,再接上銅離子,即為固定化金屬親和薄膜(immobilized metal affinity membrane, IMAM),之後再吸附固定盤尼西林醯胺酵素。 第一部分探討再生纖維素薄膜固定酵素後接著進行酵素反應的動力學研究,探討基質固定盤尼西林醯胺酵素的抑制現象,並探討產物6-APA和phenyl acetic acid(PAA)是否為競爭型反應,由實驗結果得知在高濃度盤尼西林G對固定盤尼西林醯胺酵素有抑制的情形。我們利用Lineweaver–Burk 雙倒數作圖法,求得固定化PGA酵素之動力學參數,VM =0.212 mM/min, KM= 2.55 mM, KiS= 411.29 mM,KiPAA= 6.02 mM, Ki6-APA= 1.22 mM。將參數來模擬基質PG隨時間的變化,其模擬結果實際實驗值與模擬計算值之擬合程度,證實其產物有抑制現象。 第二部分探討再生纖維素薄膜固定酵素在柱狀流動反應器中的變因如膜片數、隔板、流速,找到最適化的條件。由實驗結果可知發現當膜片數為10片且加隔板反應為最佳條件,流速則為0.58 ml/min可幾乎達到98%的轉化率。zh_TW
dc.description.tableofcontents中文摘要 i 英文摘要 ii 目錄 iii 圖目錄 v 表目錄 vii 第一章、緒論 1 第二章、文獻回顧 2 2-1固定化金屬親和層析 2 2-2薄膜應用於金屬親和層析法 8 2-3抗生素的介紹 9 2-4盤尼西林醯胺酵素 10 2-5 PGA酵素固定化 11 2-6酵素動力學 14 2-6-1 Michaelis 參數之估算 15 2-6-2動力學理論 19 2-7 酵素膜反應器(Biocatalytic membrane reactors) 24 第三章、實驗材料與方法 26 3-1實驗藥品 26 3-2儀器設備 27 3-3盤尼西林醯胺粗酵素液的製備 28 3-4固定化金屬親和薄膜之製備 28 3-5批次吸附實驗 29 3-5-1製備吸附PGA之IMAM膜 29 3-5-2基質(PG)測量 29 3-5-3產物(6-APA)測量 30 3-5-4產物(PAA)測量 30 3-6流動式吸附實驗(裝置如圖) 30 3-6-1 膜片數變數 30 3-6-2 隔板變數 30 3-6-3 不同流速變數 31 3-7分析方法 31 3-7-1 PGA酵素活性的量測 31 3-7-2金屬離子的量測 32 第四章、實驗結果與討論 34 4-1 Batch酵素動力學探討 34 4-1-1基質(PG)動力學反應 34 4-1-2產物PAA抑制反應 39 4-1-3產物6-APA抑制反應 41 4-1-4動力學模式 43 4-2 Plug flow reactor流動式探討 49 4-2-1隔板比較 49 4-2-2膜片數比較 51 4-2-3 不同流速比較 53 第五章、結論與未來展望 54 5-1結論 54 5-2未來展望 54 參考文獻 55zh_TW
dc.language.isozh_TWen_US
dc.publisher化學工程學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1508201215475000en_US
dc.subject盤尼西林醯胺酵素zh_TW
dc.subjectpenicillin G acylaseen_US
dc.subject固定化金屬親和薄膜zh_TW
dc.subject動力學zh_TW
dc.subjectImmobilized metal affinity membraneen_US
dc.subjectkineticsen_US
dc.title金屬親和薄膜固定盤尼西林醯胺酵素在批式反應器和流動式反應器的動力學探討zh_TW
dc.titleStudy of the kinetics for penicillin G acylase on immobilized metal affinity membrane for batch and plug flow reactoren_US
dc.typeThesis and Dissertationzh_TW
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item.languageiso639-1zh_TW-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
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