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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王愛玉 | |
| dc.contributor.author | Tz-Li Chen | en |
| dc.contributor.author | 陳姿利 | zh_TW |
| dc.date.accessioned | 2021-06-13T07:06:25Z | - |
| dc.date.available | 2005-07-30 | |
| dc.date.copyright | 2005-07-30 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-26 | |
| dc.identifier.citation | 參考文獻
Albrecht, G., and Mustroph, A. (2003). Localization of sucrose synthase in wheat roots: increased in situ activity of sucrose synthase correlates with cell wall thickening by cellulose deposition under hypoxia. Planta 217, 252-260. Amor, Y., Haigler, C.H., Johnson, S., Wainscott, M., and Delmer, D.P. (1995). A membrane-associated form of sucrose synthase and its potential role in synthesis of cellulose and callose in plants. Proceedings of the National Academy of Sciences of the United States of America 92, 9353-9357. Anand, S.S., R. (1986). Sucrose synthase from immature wheat grains. Journal of Plant Science Research 2, 1-10. Anguenot, R., Yelle, S., and Nguyen-Quoc, B. (1999). Purification of tomato sucrose synthase phosphorylated isoforms by Fe(III)-immobilized metal affinity chromatography. Archives of Biochemistry & Biophysics 365, 163-169. Baroja-Fernandez, E., Munoz, F.J., Saikusa, T., Rodriguez-Lopez, M., Akazawa, T., and Pozueta-Romero, J. (2003). Sucrose synthase catalyzes the de novo production of ADPglucose linked to starch biosynthesis in heterotrophic tissues of plants. Plant & Cell Physiology 44, 500-509. Barratt, D.H., Barber, L., Kruger, N.J., Smith, A.M., Wang, T.L., and Martin, C. (2001). Multiple, distinct isoforms of sucrose synthase in pea. Plant Physiology 127, 655-664. Bologa, K.L., Fernie, A.R., Leisse, A., Loureiro, M.E., and Geigenberger, P. (2003). A bypass of sucrose synthase leads to low internal oxygen and impaired metabolic performance in growing potato tubers. Plant Physiology 132, 2058-2072. Bordier, C. (1981). Phase separation of integral membrane proteins in Triton X-114 solution. Journal of Biological Chemistry 256, 1604-1607. Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248-254. Cardini, C.E., Leloir, L.F. and Chiriboga, J. (1995). The biosynthesis of sucrose. Journal of Biological Chemistry 214, 149-155. Carlson, S.J., and Chourey, P.S. (1996). Evidence for plasma membrane-associated forms of sucrose synthase in maize. Molecular & General Genetics 252, 303-310. Carlson, S.J., Chourey, P.S., Helentjaris, T., and Datta, R. (2002). Gene expression studies on developing kernels of maize sucrose synthase (SuSy) mutants show evidence for a third SuSy gene. Plant Molecular Biology 49, 15-29. Chen, Y.C. and Chourey, P.S. (1989). Spatial and temporal expression of the two sucrosesynthase genes in maize: Immunohistological evidence. Theoretical and Applied Genetics 78, 553-559. Chourey, P. S. and Nelson, O.E. (1976). The enzymatic deficiency conditioned by the shrunken-1 mutation in maize. Biochemical Genetics 14, 1041-1055. Chourey, P.S., Taliercio, E.W., Carlson, S.J., and Ruan, Y.L. (1998). Genetic evidence that the two isozymes of sucrose synthase present in developing maize endosperm are critical, one for cell wall integrity and the other for starch biosynthesis. Molecular & General Genetics 259, 88-96. Chourey. P. S. (1981). Genetic control of sucrose synthase in maize endosperm. Molecular & General Genetics 184: 372-376. Cregg, J. M., Barringer, K. J., and Hessler, A.Y., (1985). Pichia pastoris as a host system for transformations. Molecular Cell Biology 5, 3376-3385. Denyer, K., Dunlap, F., Thorbjornsen, T., Keeling, P., and Smith, A.M. (1996). The major form of ADP-glucose pyrophosphorylase in maize endosperm is extra-plastidial. Plant Physiology 112, 779-785. Dejardin, A., Sokolov, L.N., and Kleczkowski, L.A. (1999). Sugar/osmoticum levels modulate differential abscisic acid-independent expression of two stress-responsive sucrose synthase genes in Arabidopsis. Biochemical Journal 344 Pt 2, 503-509. Elling, L. (1995). Effect of metal ions on sucrose synthase from rice grains--a study on enzyme inhibition and enzyme topography. Glycobiology 5, 201-206. Etxeberria, E., and Gonzalez, P. (2003). Evidence for a tonoplast-associated form of sucrose synthase and its potential involvement in sucrose mobilization from the vacuole. Journal of Experimental Botany 54, 1407-1414. Geigenberger, P. and Stitt, M. (1993). Sucrose synthase catalyses a readily reversible reaction in vivo in developing potato tubers and other plant tissues. Planta 189, 329-339. Gross, K.C.P., D.M. (1982). Cucumber fruit sucrose synthetase isozymes. Phytochemistry 21. Guilbault, George G. (1976) Handbook of Enzyme Methods of Analysis. Hanggi, E., and Fleming, A.J. (2001). Sucrose synthase expression pattern in young maize leaves: implications for phloem transport. Planta 214, 326-329. Hardin, S.C., Winter, H., and Huber, S.C. (2004). Phosphorylation of the amino terminus of maize sucrose synthase in relation to membrane association and enzyme activity. Plant Physiology 134, 1427-1438. Hardin, S.C., Tang, G.Q., Scholz, A., Holtgraewe, D., Winter, H., and Huber, S.C. (2003). Phosphorylation of sucrose synthase at serine 170: occurrence and possible role as a signal for proteolysis. Plant Journal 35, 588-603. Hisajima, S.I., T. (1981). Purification and properties of sucrose synthetase from morning glory callus cells. Biologia Plantarum 23. Huang, D.Y., and Wang, A.Y. (1998). Purification and characterization of sucrose synthase isozymes from etiolated rice seedlings. Biochemistry & Molecular Biology International 46, 107-113. Huang, J.W., Chen, J.T., Yu, W.P., Shyur, L.F., Wang, A.Y., Sung, H.Y., Lee, P.D., and Su, J.C. (1996). Complete structures of three rice sucrose synthase isogenes and differential regulation of their expressions. Bioscience, Biotechnology & Biochemistry 60, 233-239. Huang, Y.H., Picha, D.H., and Kilili, A.W. (1999). A continuous method for enzymatic assay of sucrose synthase in the synthetic direction. Journal of Agricultural & Food Chemistry 47, 2746-2750. Hubbard, N. L., Huber, S. C., and Pharr, D. M. (1989). Sucrose phosphate synthase and acid invertase as determinants of sucrose concentration in developing muskmelon (Cucumis melo L.) fruits. Plant Physiololgy 91, 1527-1534. Huber, S.C., Huber, J.L., Liao, P.C., Gage, D.A., McMichael, R.W., Jr., Chourey, P.S., Hannah, L.C., and Koch, K. (1996). Phosphorylation of serine-15 of maize leaf sucrose synthase. Occurrence in vivo and possible regulatory significance. Plant Physiology 112, 793-802. Koch, K.E., Nolte, K.D., Duke, E.R., McCarty, D.R., and Avigne, W.T. (1992). Sugar Levels Modulate Differential Expression of Maize Sucrose Synthase Genes. Plant Cell 4, 59-69. Koch, K. E., Xu, J., Duke, E. R., Yuan, C.-X., Tan, B.-C., and Avigne, W. T. (1995). Sucrose provides long distance signal for coarse control of gene aggecting its metabolism. In: Sucrose metabolism, Biochemistry, Physiology and Molecular Biology, Point, H. G., Salerno, H.G., and Echeverria, E., Eds., American Society of Plant Physiologists., Rockville, MD, 266-277. Koch, K. (2004). Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Current Opinion in Plant Biology 7, 235-246. Komina, O., Zhou, Y., Sarath, G., and Chollet, R. (2002). In vivo and in vitro phosphorylation of membrane and soluble forms of soybean nodule sucrose synthase. Plant Physiology 129, 1664-1673. Konishi, T., Ohmiya, Y., and Hayashi, T. (2004). Evidence that sucrose loaded into the phloem of a poplar leaf is used directly by sucrose synthase associated with various beta-glucan synthases in the stem. Plant Physiology 134, 1146-1152. Larsen, A.E., Salerno, G.L. and Pontis, H.G. (1985). Sucrose synthase from wheat leaves. Comparison with the wheat germ enzyme. Physiologia Plantarum 67, 37-42. Lindblom, S., Ek, P., Muszynska, G., Ek, B., Szczegielniak, J., and Engstrom, L. (1997). Phosphorylation of sucrose synthase from maize seedlings. Acta Biochimica Polonica 44, 809-817. Macauley-Patrick, S., Fazenda, M.L., McNeil, B., and Harvey, L.M. (2005). Heterologous protein production using the Pichia pastoris expression system. Yeast 22, 249-270. Martin, T., Frommer, W.B., Salanoubat, M., and Willmitzer, L. (1993). Expression of an Arabidopsis sucrose synthase gene indicates a role in metabolization of sucrose both during phloem loading and in sink organs. Plant Journal 4, 367-377. Matic, S., Akerlund, H.E., Everitt, E., and Widell, S. (2004). Sucrose synthase isoforms in cultured tobacco cells. Plant Physiology & Biochemistry (Paris) 42, 299-306. Morell, M.C., L. (1985). Sucrose synthase of soybean nodules. Plant Physiology 78. Nakai, T., Moriya, A., Tonouchi, N., Tsuchida, T., Yoshinaga, F., Horinouchi, S., Sone, Y., Mori, H., Sakai, F., and Hayashi, T. (1998a). Control of expression by the cellulose synthase (bcsA) promoter region from Acetobacter xylinum BPR 2001. Gene 213, 93-100. Nakai, T., Konishi, T., Zhang, X.Q., Chollet, R., Tonouchi, N., Tsuchida, T., Yoshinaga, F., Mori, H., Sakai, F., and Hayashi, T. (1998b). An increase in apparent affinity for sucrose of mung bean sucrose synthase is caused by in vitro phosphorylation or directed mutagenesis of Ser11. Plant & Cell Physiology 39, 1337-1341. Nakai, T., Tonouchi, N., Konishi, T., Kojima, Y., Tsuchida, T., Yoshinaga, F., Sakai, F., and Hayashi, T. (1999). Enhancement of cellulose production by expression of sucrose synthase in acetobacter xylinum. Proceedings of the National Academy of Sciences of the United States of America 96, 14-18. Nelson, N. (1944). A photometric adaption Somogi method for the determination of glucose . Journal of Biological Chemistry 153, 375-380. Patil, B.A.J., G.V. (1972). Effect of magnesium(2+) and manganese(2+) ions on sucrose synthetase in sugarcane leaves. Proceedings of the Indian National Science Academy. Part B 38, 50-54. Pozueta-Romero, J., Yamaguchi, J., and Akazawa, T. (1991). ADPG formation by the ADP-specific cleavage of sucrose-reassessment of sucrose synthase. FEBS Letters 291, 233-237. Pozueta-Romero, J., Perata, P. and Akazawa, T. (1999). Sucrose-starch conversion in heterotropic tissues of plants. Critical Reveiws in Plant Science 18, 489-525. Romer, U., Schrader, H., Gunther, N., Nettelstroth, N., Frommer, W.B., and Elling, L. (2004). Expression, purification and characterization of recombinant sucrose synthase 1 from Solanum tuberosum L. for carbohydrate engineering. Journal of Biotechnology 107, 135-149. Ricard, B., Toai T. V., Chourey P., and Saglio P. (1998). Evidence for the critical role of sucrose synthase for anoxic tolerance of maize roots using a double mutant. Plant Physiol 116 (4), 1323-1331. Ruan, Y.L., Llewellyn, D.J., and Furbank, R.T. (2003). Suppression of sucrose synthase gene expression represses cotton fiber cell initiation, elongation, and seed development. Plant Cell 15, 952-964. Ruan, Y.L., Llewellyn, D.J., Furbank, R.T., and Chourey, P.S. (2005). The delayed initiation and slow elongation of fuzz-like short fibre cells in relation to altered patterns of sucrose synthase expression and plasmodesmata gating in a lintless mutant of cotton. Journal of Experimental Botany 56, 977-984. Salnikov, V.V., Grimson, M.J., Seagull, R.W., and Haigler, C.H. (2003). Localization of sucrose synthase and callose in freeze-substituted secondary-wall-stage cotton fibers. Protoplasma 221, 175-184. Salanoubat, M., and Belliard, G. (1989). The steady-state level of potato sucrose synthase mRNA is dependent on wounding, anaerobiosis and sucrose concentration. Gene 84, 181-185. Sayion, Y., Huang, Y. W., Chen H. W., Liao Y. C. and Wang A. Y. (1999). Expression and characterization of rice sucrose synthase in Escherichia coli. Food Science and Agricultural Chemistry 1(2), 122-128. Shaw, J.R., Ferl, R.J., Baier, J., St.Clair, D., Carson, C. and McCarty, L.C. (1994). Structural features of maize Sus1 gene and protein. Plant Physiology 106, 1659-1665. Sung, H.Y. and Su, J.C. (1977). Sucrose synthase isozymes of pea seedling - Purification and general properties. Journal Chinese Biochemisyry Society 6, 22-37. Su, J. C., Wu J. L., and Yang, C. L. (1977). Purification and characterization of sucrose synthetase from the shoot of bamboo Leleba oldhami. Plant Physiology 60, 17-21. Su, J.C. and Preiss, J. (1978). Purification and properties of sucrose synthase from maize. Plant Physiology 61, 389-393. Tanase, K., and Yamaki, S. (2000). Purification and characterization of two sucrose synthase isoforms from Japanese pear fruit. Plant & Cell Physiology 41, 408-414. Tang, G.Q., and Sturm, A. (1999). Antisense repression of sucrose synthase in carrot (Daucus carota L.) affects growth rather than sucrose partitioning. Plant Molecular Biology 41, 465-479. Tsai Z. C. and Wang A. Y. (2003). Identification of rice manganese-dependent protein kinases that phosphorylate sucrose synthase at multiple serine residues. Botanical Bulletin of Academia Sinica 44,141-150. Vinci, J., Nolte, K.D. and Koch, K.E. (1993). Sucrose synthase localization during seed and hair development: Cotton ovules as a model system. Plant Physiology 102, 43-43 Wacher R, Langhans M, Aloni R, Gotz S, Weilmunster A, Koops A, Temguia L, Mistrik I, Pavlovkin J, Rascher U et al. (2003). Vascularization, high-volumn solution flow, and localized roles for enzymes of sucrose metabolism during tumorigenensis by Agrobacterium tumefaciens. Plant Physiology 133, 1024-1037. Wang, A.Y., Yu, W.P., Juang, R.H., Huang, J.W., Sung, H.Y., and Su, J.C. (1992). Presence of three rice sucrose synthase genes as revealed by cloning and sequencing of cDNA. Plant Molecular Biology 18, 1191-1194. Wang, A.Y., Kao, M.H., Yang, W.H., Sayion, Y., Liu, L.F., Lee, P.D., and Su, J.C. (1999). Differentially and developmentally regulated expression of three rice sucrose synthase genes. Plant & Cell Physiology 40, 800-807. Wang, F., Smith, A.G. and Brenner, M.L. (1994). Temporal and spatial expression pattern of sucrose synthase during tomato fruit development. Plant Physiology 104, 535-540. Winter, H., and Huber, S.C. (2000). Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes. Critical Reviews in Biochemistry & Molecular Biology 35, 253-289. Winter, H., Huber, J.L., and Huber, S.C. (1997). Membrane association of sucrose synthase: changes during the graviresponse and possible control by protein phosphorylation. FEBS Letters 420, 151-155. Winter, H., Huber, J.L., and Huber, S.C. (1998). Identification of sucrose synthase as an actin-binding protein. FEBS Letters 430, 205-208. Wolfgang E. Schafer, Rohwerb, J.M., and Bothac, F.C. (2004). Protein-level expression and localization of sucrose synthase in the sugarcane culm. Physiologia Plantarum 121, 187–195. Wolosiuk, R.A. and Pontis, H.G. (1971). Evidence of the existence of two forms of sucrose synthetase. FEBS Letters 16, 237-240. Yen, S.F., Su, J.C., and Sung, H.Y. (1994). Purification and characterization of rice sucrose synthase isozymes. Biochemistry & Molecular Biology International 34, 613-620. Zhang, X.Q., and Chollet, R. (1997). Seryl-phosphorylation of soybean nodule sucrose synthase (nodulin-100) by a Ca2+-dependent protein kinase. FEBS Letters 410, 126-130. Zrenner, R., Salanoubat, M., Willmitzer, L., and Sonnewald, U. (1995). Evidence of the crucial role of sucrose synthase for sink strength using transgenic potato plants (Solanum tuberosum L.). Plant Journal 7, 97-107. 莊榮輝 (1985) 水稻蔗糖合成酶之研究－其純化、生物化學及免疫性質之比較。博士論文，國立台灣大學農業化學研究所。 黃如瑋 (1994) 水稻蔗糖合成酶異構基因之選殖及影響表現生理條件的研究。博士論文，台灣大學農業化學研究所。 岩素芬 (1992) 水稻蔗糖合成異構酶之純化與鑑定。博士論文，國立台灣大學農業化學研究所。 廖憶純 (2002) 水稻懸浮培養細胞中蔗糖合成酶基因表現受糖調控之研究。博士論文，國立台灣大學農業化學研究所。 蔡承佳 (2003) 蛋白質磷酸化對水稻蔗糖合成酶酵素功能及基因表現的影響。博士論文，國立台灣大學農業化學研究所。 黃德宜 (2003) 水稻蔗糖合成酶 RSuS3 基因表現與酵素功能之探討。博士論文，國立台灣大學農業化學研究所。 莊榮輝 (1997) 酵素化學實驗 – 實驗手冊。 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/35716 | - |
| dc.description.abstract | 蔗糖合成酶催化蔗糖與 UDP 轉換為果糖及 UDPG 之可逆反應。在水稻中有三種 RSus 基因 (RSus1, RSus2, Rus3)，RSus1 以及RSus2 在水稻中各組織中皆有表現，而 RSus3 則主要表現於水稻種子中。本論文由 Pichia pastoris 表現之 RSuS2，探討其酵素功能。
將 RSus2 cDNA 送入酵母菌 P. pastoris 中表現，轉形株以甲醇誘導 60 小時後，可以得到最大的 RSuS2 蛋白質量以及 SuS 活性。將重組 RSuS2 進行純化並進行生化性質分析。重組 RSuS2 對於蔗糖、UDP、果糖及UDPG之 Km 值分別為 104.8 mM、0.099 mM、6.65 mM 及 0.146 mM。Ca2+ 以及 Mg2+ 會促進合成方向的活性。代謝中間產物 fructose 6-phosphate 會抑制分解方向的活性，而 fructose 2,6 bisphosphate 則會活化分解方向的活性。此外，重組 RSuS2 具有膜結合型式的蛋白質存在，並且純化後的重組 RSuS2 分解方向活性可以被界面活性劑 Triton X-100 活化，合成方向的活性則可以被磷脂質 (phospholipid) 活化。 | zh_TW |
| dc.description.abstract | Sucrose synthase (SuS) catalyzes the reversible conversion of sucrose and UDP into fructose and UDPG. There are three Sus isogenes in rice (Oryza sativa) (RSus1, RSus2 and RSus3). Among these three isogenes, RSus1 and RSus2 are ubiquitously expressed in various tissues, while RSus3 is expressed predominantly in rice seed. In this research, recombinant RSuS2 expressed and purified from Pichia pastoris was used to study on the enzyme function of RSuS2.
The RSus2 cDNA was introduced into yeast Pichia pastoris for expression. The transformed cells accumulated high level of RSuS2 protein and SuS activity after 60 hours of methanol induction. The recombinant RSuS2 was purified and characterized on its general properties. The Michaelis constant (Km) of the recombinant RSuS2 for sucrose, UDP, fructose, and UDPG were 104.8 mM, 0.099 mM, 6.65 mM, and 0.146 mM, respectively. The synthetic activity could be stimulated by Mg2+ and Ca2+. Moreover, the cleavage activity of RSuS2 was inhibited by fructose 6-phosphate, but was activated by fructose 2,6 bisphosphate. On the other hand, membrane-bound RSuS was observed in the crude extract of recombinant RSuS2. The cleavage activity of purified RSuS2 was activated by Triton X-100, while the synthetic activity was activated by phospholipids. | en |
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| dc.description.tableofcontents | 目錄………………………………………………………….……..I
寫表…………………………………………………………………...V 摘要…………………………………………………………………..VI Abstract…………………………………………………………….VII 第一章 研究背景…………………………………………………..1 第一節 蔗糖合成酶的生化性質…………………………………….1 第二節 蔗糖合成酶的生理功能…………………………………….2 2.1 不同型式之蔗糖合成酶 2 2.2 參與細胞壁多醣類的合成 3 2.3 參與澱粉的生合成 4 2.4 參與蔗糖運輸 4 2.5 缺氧逆境 5 第三節 蔗糖合成酶的磷酸化現象…………………………………5 第四節 蔗糖合成酶之基因調控…………………………………….6 第五節 本篇論文之目的及方向…………………………………….7 6.1 重組 RSuS2 於酵母菌的表現及純化 8 6.2 重組 RSuS2 生化性質之探討 8 第二章 材料與方法…………………………………………………10 第一節 實驗材料與樣品………………………………………10 1.1 菌種 10 1.2 質體 10 1.3 藥品 10 第二節 實驗儀器設備………………………………………………10 2.1 水平核酸電泳 10 2.2 蛋白質電泳轉印設備 10 2.3 離心機 10 2.4 其他 11 第三節 實驗方法……………………………………………………11 3.1質體 DNA 抽取與分析 11 3.1.1質體 DNA 之小量分離 11 3.1.2 DNA 限制酶切割 12 3.1.3 DNA 洋菜膠體電泳法 12 3.2 於酵母菌Pichia pastoris中表現重組蛋白質 12 3.2.1 酵母菌轉形 12 3.2.2 酵母菌轉形株的篩選與鑑定 13 3.3 蛋白質定量與分析 14 3.3.1 蛋白質定量 14 3.3.2蛋白質電泳 14 3.3.3 蛋白質轉印 15 3.3.4 免疫染色法 15 3.4 蔗糖合成酶活性測定 16 3.4.1 分解蔗糖的活性測定 16 3.4.1.1 UDPG 去氫酶酵素耦合法 16 3.4.1.2 連續酵素耦合測定法 17 3.4.1.3 還原糖定量法 17 3.4.2 合成蔗糖的活性測定 18 3.4.2.1 Anthrone 定量法 18 3.4.2.2 連續酵素耦合測定法 19 3.5 重組 RSuS2 蛋白質之表現及純化 20 3.5.1 重組 RSuS2 之最佳表現時間探討 20 3.5.2 重組 RSuS2 之純化 20 3.6 重組 RSuS2 生化性質分析 22 3.6.1 最適反應溫度測定 22 3.6.2 最適反應pH值測定 22 3.6.3 熱穩定性測定 23 3.6.4 pH穩定性測定 23 3.6.5 不同金屬離子的影響 23 3.6.6 不同中間代謝物的影響 23 3.6.7 無機磷酸根之影響 23 3.6.8 Phospholipid 對活性的影響 23 3.6.9 酵素動力學測定 24 3.7 其他實驗法 25 3.7.1 Triton X-114 partition 25 3.7.2 膜結合蛋白質之萃取 26 3.7.3 Glucose 6-phosphate 脫氫酶活性測定 26 第三章 結果與討論………………………………………………27 第一節 RSuS2 cDNA 於酵母菌 Pichia pastoris 中表現…………27 1.1 表現載體之轉形 27 1.2 轉形株之鑑定與表現篩選 27 1.3 重組 RSuS2蛋白質的純化 28 第二節 重組蛋白質分子量檢定………………………………………29 2.1 蛋白質電泳法 29 2.2 膠體過濾法 30 第三節 重組蛋白質 N 端定序………………………………………30 第四節 重組蛋白質的生化性質分析…………………………………30 4.1 活性測定方法之比較 30 4.2 酵素動力學常數的測定 31 4.3 最適反應溫度 32 4.4 最適反應 pH 值 32 4.5 熱穩定性 32 4.6 pH 穩定性 32 4.7 不同金屬離子的影響 33 第五節 中間代謝物對重組 RSuS2 的影響…………………………33 5.1 Hexose phosphates 的影響 34 5.2 Fructose 1,6 bisphosphate 以及 fructose 2,6 bisphosphate 的影響 34 5.3 Dihydroacetone phosphate 及 3-phosphoglycerate 的影響 34 5.4 無機磷酸根的影響 34 5.5 討論 35 第六節 重組 RSuS2 在酵母菌中膜結合型式與水溶性型式的分佈…………35 6.1 活性分佈 35 6.2 蛋白質電泳分析 36 第七節 界面活性劑以及 Phospholipid 對重組 RSuS2 的影響…37 7.1 Triton X-114 分層 37 7.2 Triton X-100 以及 phospholipid 對活性的影響 37 7.3 討論 37 第四章 總論………………………………………………………39 第五章 未來展望……………………………………………………41 5.1 RSuS2 在環境中糖濃度的改變與膜結合型式的分佈 41 5.2 RSuS2 與膜結合之區域 41 5.3 Fructose 6-phsphate 及 fructose 2,6 bisphaphate 對 RSuS2 活性影響的情形 41 參考文獻……………………………………………………………….42 圖表…………………………………………………………………….49 | |
| dc.language.iso | zh-TW | |
| dc.subject | 水稻蔗糖合成酶 | zh_TW |
| dc.subject | SuS | en |
| dc.subject | sucrose synthase | en |
| dc.subject | Pichia pastoris | en |
| dc.title | 水稻蔗糖合成酶 RSuS2 在酵母菌 Pichia pastoris 中的表現及生化性質檢定 | zh_TW |
| dc.title | Expression and Characterization of Rice Sucrose Synthase RSuS2 in Pichia pastoris | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 宋賢一,蔣啟玲,張珍田 | |
| dc.subject.keyword | 水稻蔗糖合成酶, | zh_TW |
| dc.subject.keyword | sucrose synthase,SuS,Pichia pastoris, | en |
| dc.relation.page | 77 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-27 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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