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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.author | Chen-Chen Tu | en |
dc.contributor.author | 涂真真 | zh_TW |
dc.date.accessioned | 2021-07-01T08:17:13Z | - |
dc.date.available | 2021-07-01T08:17:13Z | - |
dc.date.issued | 1993 | - |
dc.identifier.citation | (1) Enzymes Nomenclature, Academic Press, New York, 1979.
(2) S.T. Chen & K.T. Wang, “The Synthesis of β-Benzyl L-Aspartate and γ-Benzyl L-Glutamate by Enzyme-Catalyzed Hydrolysis”, Synthesis, 1987, 581-582. (3) M.J. Daniel, “Industrial Operation of Immobilized Enzyme”, Methods in Enzymology, 1987, 136, 37 1-379. (4) E.L. Smith, R.J. DeLange, W.H. Evans, M. Landon & F.S. Markland, “Subtilisin Carlsberg (V. The Complete Sequence; Comparison with Subtilisin BPN’; Evolutionary Relationships)”, J. Biol. Chem. 1968, 243, 2184-2191. (5) MJ. Daniel, “Industrial Operation of Immobilized Enzyme”, Methods in Enzymology, 1987, 136, 371-379. (6) K. Veeraragavan, T. Colpitts & B.F. Gibbs, “Purification and Characterization of Two Distinct Lipases from Geotrichum Candidum”, Biochimica et Biophysica Acta, 1990, 1044, 26-33. (7) Availible from NOVO Industri A/S as a clear brown liquid. According to NOVO, one Anson Unit (AU) is the amount of enzyme which, under standard conditions digests haemoglobin at an initial rate liberating per mm an amount of TCA-soluble product which gives the same colour with phenol reagent as 1 mequiv. of tyrosine (1AU = lOOμMol of Tyr-OMe hydrolysed per min). Thus Alacase 2.5L (trade name of NOVO Industri) contains 2.5 Aug-1. (8) S.T. Chen, K.T. Wang & C.H. Wong, “Chirally Selective Hydrolysis of D,L-Amino Acid Esters by Alkaline Protease”, J. Chem. Soc., Chem. Commun. 1986, 1514-1516. (9) S.T. Chen, S.H. Wu & K.T. Wang, “Diastereoselective Hydrolysis of Peptide Esters by Alkaline Protease”, Int. J. Peptide Protein Res. 1991, 37, 347-350. (10) S.T. Chen, S.Y. Chen & K.T. Wang, “Kinetically Controlled Peptide Bond Formation in Anhydrous Alcohol Catalyzed by an Industrial Protease ‘Alcalase”, J. Org. Chem. 1992, 57, 6960-6965. (11) S.T. Chen & K.T. Wang, “Enzyme-Catalysed Selective Ester Hydrolysis of Aspartyl and Glutamyl Dipeptide Benzyl Esters”, J. Chem. Research(S), 1987, 308-309. (12) A. Zaks & A.M. Klibanov, “Enzymatic Catalysis in Organic Media at 100°C”, Science, 1984,224, 1249-1251. (13) T. Sakurai, A.L. Margolin, A.J. Russell & A.M. Klibanov, “Control of Enzyme Enantioselectivity by the Reaction Medium”, J. Am. Chem. Soc. 1988, 110, 7236-7237. (14) G.A. Homandberg, J.A. Mattis & M. Laskowski, Jr., “Synthesis of Peptide Bonds by Proteinases. Addtion of Organic Cosolvents Shifts Peptide Bond Equilibria toward Synthesis”, Biochemistry, 1978, 17, 5220-5227. (15) S.T. Chen, S.Y. Chen, S.C. Hsiao & K.T. Wang, “Application of Industrial Protease ‘Alcalase’ in Peptide Synthesis”, Biomed. Biochim. Acta,1991,50, 10/11, 181-186. (16) T.L. Chapman, T.B. Shull & F.M. Raushel, “Stereochemical Probes of the Argininosuccinate Synthetase Reaction”, Biochemistry, 1986, 25, 4739-4744. (17) D. Halliday, 1988, “Recent advances in the use of labelled amino-acids to quantative protein metabolism in man” In Synthesis and Applications of Isotopically Labelled Compounds. Proceedings of the Third International Symposium, Innsbruck, Austria 1988, p.177-182, Eds. T.A. Baillie, I.R. Jones, 1989, Elsevier Science Publishers B.V., Amsterdam. (18) D.S. Millington, D.A. Maltby, D. Gale, C.R. Roe, 1988, “Synthesis and human applications of stable isotope-labelled L-carnitine” Synthesis and Aplications of Isotopically Labelled Compounds. Proceedings of the Third International Symposium, Innsbruck, Austria 1988, p.189-1949 Eds. T.A. Baillie, I.R. Jones, 1989, Elsevier Science Publishers B.V., Amsterdam. (19) K.A.H. Adams, S.H. Chung & A.M. Klibanov, “Kinetic Isotope Effect Investigation of Enzyme Mechanism in Organic Solvents”, J. Am. Chem. Soc. 1990, 112, 94 18-9419. (20) C.H. Wong & G.M. Whitesides, “Enzyme-Catalyzed Organic Synthesis:Regeneration of Deuterated Nicotinamide Cofactors for Use in Large-Scale Enzymatic Synthesis of Deuterated Substances”, J. Am. Chem.Soc. 1983, 105, 5012-5014. (21) E. Gout, S. Chesne, C.G. Beguin & J. Pelmont, “Kinetic Studies with the Use of Proton-Magnetic-Resonance Spectroscopy of the Specific α-Deuteration of Amino Acids by Escherichia coli Aspartate Aminotransferase”, Biochem. J. 1978, 171, 719-723. (22) D.M. Yamamoto, D.A. Upson, D.K. Linn & V.J. Hruby, “Synthesis of Specific Deuterium Labeled Tyrosine and Phenylalanine Derivatives and Their Use in the Total Synthesis of [8-Arginine]vasopressin Derivatives:The Separation of Diastereomeric [8-Argininejvasopressin Derivatives by Partition Chromatography”, J. Am. Chem. Soc. 1977, 99, 1564-1570. (23) S.T. Chen, S.Y. Chen, S.C. Hsiao & K.T. Wang, “Kinetic Resolution of N-Protected Amino Acid Esters in Organic Solvents Catalyzed by a Stable Industrial Alkaline Protease”, Biotechnology Letters, 1991, 13, 773-778. (24) L.G. Barry, M. Pugniere, B. Castro & A. Previero, “Racemization ofα-Amino Acid Esters by Aliphatic Ketones in the Presence of Carboxylic Acids”, Int. J. Peptide Protein Res. 1993, 41, 323-325. (25) H. Kitaguchi & A.M. Klibanov, “Enzymatic Peptide Synthesis via Segment Condensation in the Presence of Water Mimics”, J. Am. Chem. Soc. 1989, 111, 9272-9273. (26) P.A. Fitzpatrick & A.M. Klibanov, “How Can the Solvent Affect Enzyme EnantioselectivityT’, J. Am. Chem. Soc. 1991, 113, 3166- 3171. (27) H. Kitaguchi, I. Itoh & M. Ono, “Effects of Water and Water- Mimicking Solvents on the Lipase-Catalyzed Esterification in an Apolar Solvent”, Chemistry Letters, 1990, 1203-1206. (28) J.A. Wells & D.A. Estell, “Subtilisin - an Enzyme Designed to be Engineered”, T.I.B.S. 1988, 13, 291-297. (29) M.M. Bradford, “A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding”, Analytical Biochemistry, 1976, 72, 248-254. (30) M. Tal, A. Silberstein & E. Nusser, “Why Does Coomassie Brilliant Blue R Interact Differently with Different Proteins”, J. Biol. Chem. 1980, 260, 9976-9980. (31) P. Aldercreutz & B. Mattiasson, “Aspects of Biocatalyst Stability in Organic Solvents”, Biocatalysis, 1987, 1, 99-108. (32) B. Schulze & A.M. Klibanov, “Inactivation and Stabilization of Subtilisins in Neat Organic Solvents”, Biotechnology and Bioengineering, 1991, 38, 1001-1006. (33) B.S. Hartley &B .A. Kilby, “The Inhibition of Chymotrypsin by Diethyl p-Nitrophenyl Phosphate”, Biochem. J. 1952, 50, 672-678. (34) K. Morihara, T. Oka & H. Tsuzuki, “Comparative Study of Various Serine Proteinases from Microorganisms: Specificity with Oligopeptides”, Arch. Biochem. Biophys. 1971, 146, 297-305. (35) J.B. Jones, “Enzymes in Organic Synthesis”, Tetrahedron, 1986, 42, 3351-3403. (36) A.M. Klibanov, “Enzymes That Work in Organic Solvents”, Chemtech. 1986, Jane, 354-359. (37) C.H. Wong, “Enzymatic Catalysts in Organic Synthesis”, Science, 1989, 244, 1145-1152. | - |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76010 | - |
dc.description.abstract | Alcalase 是枯草桿菌蛋白酵素的其中一種,為自 Bacillus lichenifomis 菌株篩選得到的鹼性蛋白水解酵素。由於價格便宜,取得容易,再加上具有很廣泛的專一性(Specificity ) ,特別是對於胺基酸基質具有比較多的選擇性( Selectivity ) ,因此,利用其高效率之蛋白質水解能力在工業上被普遍的應用來充當去汙劑、清潔劑。
有鑑於酵素應用於有機合成之範疇日益廣泛。故吾人致力於探討酵素在有機溶劑中的性質及其應用。而由本人這兩年的研究結果得知: ( l ) 應用枯草桿菌蛋白酵素,可以很簡便的製備帶有同位素的單一光學胺基酸且其純度極高 。 (2) 枯草桿蛋白酵素在三級丁醇中,對 N 端未保護的胺基酸酯也具有相當高的鏡像選擇性。則基於產物( L 型胺基酸)的低溶解度,使純化步驟可直接以簡單方便的離心方式達成。 ( 3) 應用枯草桿蛋白酵素能夠有選擇性的將消旋胺基酸酯中的 D 型異構物與另一 N 端保護的 L 型胺基酸酯形成雙勝汰。此合成過程不但速率快,產量高又不必擔心會有內消旋或副反應的發生。 (4) Alcalase 與 subtilisin Carlsberg酵素活性確實會因無水乙醇中含水量的增加而加快。 ( 5 ) Alcalase 與subtilisin Carlsberg 確實會溶於無水乙醇中,且其溶解度會隨著無水乙醇中水份的增加而增加。 (6) 溶解於無水乙醇中的 Alcalase 與 subtilisin Carlsberg 在放置 150 時後,仍存有 10 %以上的酵素活性。 懸浮在無水乙醇中的Alcalase 與 subtilisin Carlsberg 其半生期分別為42時及20小時。 離心後位於澄清液中的 Alcalase ,當其靜置在含有不同水份的乙醇溶液中時,酵素穩定度會隨著水的含量增加而越愈差。 離心後位愈沈澱物中的 Alcalase ,當其靜置在含有不同水份的乙醇溶液中時,酵素活性似乎與水的含量並沒有關聯。 ( 7 ) 在有機溶劑系統中滴定酵素活性中心數目是可行的。 ( 8 ) 經由分光光度計及氣相層析儀的分析結果,得知 Alcalase 酵素分子表面並沒有一層水膜而 subtilisin Carlsberg 酵素分子表面卻需要一層水膜來維持其催化活性。 ( 9 ) 經由鏡像選擇性的分析及 Km , Kcat 比值的大小,再配合上光散射儀器的數據結果,吾人確知酵素在有機溶劑中與在緩衝液中的三級結構可以說是相同的。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-07-01T08:17:13Z (GMT). No. of bitstreams: 0
Previous issue date: 1993 | en |
dc.description.tableofcontents | 摘要. . . . . . . . . . . . . 1
( A )緒言. . . . . . . . . . . . . 4 ( B )藥品與儀器 . . . . . . . . . . . . . 22 ( C )用工業級酵素來製備帶有同位素標記之單一光學活性的異構胺基酸 (一)前言. . . . . . . . . . . . . 25 (二)試驗步驟. . . . . . . . . . . . . 27 (三)實驗結果. . . . . . . . . . . . . 32 (四)討論. . . . . . . . . . . . . 34 (五)附圖. . . . . . . . . . . . . 35 ( D )在高濃度的有機醇中用枯草桿菌蛋白酵素對 N 端未保護的消旋胺基酸酯類作鏡像分割 (一)前言. . . . . . . . . . . . . 45 (二)實驗步驟. . . . . . . . . . . . . 46 (三)實驗結果. . . . . . . . . . . . . 51 (四)討論. . . . . . . . . . . . . 52 (五)附圖. . . . . . . . . . . . . 53 ( E )在有機醇中利用枯草桿菌蛋白酵素以動力控制法(kinetic controlled method )來選擇性的將消旋異構物 中的 D 型胺基酸酯類與蛋白質形成勝汰鍵 (一)前言- - - - - - - - - - - - - - - - - - - - - - - 61 (二)實驗步驟- - - - - - - - - - - - - - - - - - - - 63 (三)實驗結果- - - - - - - - - - - - - - - - - - - - 70 (四)討論- - - - - - - - - - - - - - - - - - - - - - - 72 (五)附圖- - - - - - - - - - - - - - - - - - - - - - - 75 ( F ) 探討無水乙醇中水的含量對 Alcalase 與 Carlsberg 反應速率的影響 (一)前言- - - - - - - - - - - - - - - - - - - - - - -83 (二)實驗步驟- - - - - - - - - - - - - - - - - - - -84 (三)實驗結果- - - - - - - - - - - - - - - - - - - -87 (四)討論- - - - - - - - - - - - - - - - - - - - - - -89 ( G ) 測試 Alcalase與 Carlsberg 在無水乙醇中的溶解度並探討其溶解度與無水乙醇中含水量的關係 (一)前言- - - - - - - - - - - - - - - - - - - - - - -90 (二)實驗步驟- - - - - - - - - - - - - - - - - - - 92 (三)實驗結果- - - - - - - - - - - - - - - - - - - 100 (四)討論- - - - - - - - - - - - - - - - - - - - - - -105 ( H )探討 Alcalase 與 Carlsberg 在無水乙醇中的穩定性 (一)前言- - - - - - - - - - - - - - - - - - - - - - 106 (二)實驗步驟- - - - - - - - - - - - - - - - - - - 109 (三)實驗結果- - - - - - - - - - - - - - - - - - - 113 (四)討論- - - - - - - - - - - - - - - - - - - - - - -117 ( I ) 測試在有機溶劑系統中滴定酵素活化中心數目的可行性 (一)前言- - - - - - - - - - - - - - - - - - - - - -120 (二)實驗步驟- - - - - - - - - - - - - - - - - - - 121 (三)實驗結果- - - - - - - - - - - - - - - - - - - 124 (四)討論- - - - - - - - - - - - - - - - - - - - - - 129 ( J ) 測試枯草桿菌蛋白酵素表面是否存在一層水膜 (一)前言- - - - - - - - - - - - - - - - - - - - - -130 (二)實驗步驟- - - - - - - - - - - - - - - - - - - - 131 (三)實驗結果- - - - - - - - - - - - - - - - - - - - 134 (四)討論- - - - - - - - - - - - - - - - - - - - - - 137 ( K ) 枯草桿菌蛋白酵素在無水溶劑中是否能維持其特異的選擇性及其三級結構 (一)前言- - - - - - - - - - - - - - - - - - - - - -138 (二)實驗步驟- - - - - - - - - - - - - - - - - - - 140 (三)實驗結果- - - - - - - - - - - - - - - - - - - 146 (四)討論- - - - - - - - - - - - - - - - - - - - - -150 (五)附圖- - - - - - - - - - - - - - - - - - - - - - 152 ( L ) 總結- - - - - - - - - - - - - - - - - - - - - - 189 ( M ) 參考資料- - - - - - - - - - - - - - - - - - -195 ( N ) 附錄- - - - - - - - - - - - - - - - - - - - - - 200 | - |
dc.language.iso | zh-TW | - |
dc.title | 枯草桿菌蛋白酵素在有機合成反應之應用及其在乙醇中特殊性質之研究 | zh_TW |
dc.title | The Application of Alcalase in Organic Synthesis and The Study of Alcalase in Alcohol | en |
dc.date.schoolyear | 81-2 | - |
dc.description.degree | 碩士 | - |
dc.relation.page | 225 | - |
dc.rights.note | 未授權 | - |
dc.contributor.author-dept | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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