臺灣博碩士論文加值系統

綠竹筍 (Leleba oldhami) 中除可溶性蔗糖合成 (Sucrose synthase，簡稱 SuS) 外，亦發現膜結合型 SuS 存在。此種結合型 SuS 含量少，但比活性很高。結合型 SuS 與細胞膜之間作用力弱，暗示其應屬於周邊性膜蛋白。可溶性SuS 經硫酸銨分劃、膠體過濾層析 (Sephacryl S-300)、陰離子交換層析 (DEAE-Sephacel) 等步驟，可得部分純化酵素。SuS 經色層焦集層析 (Mono-P chromatofocusing chromatography) 之進一步純化後發現，竹筍中至少含有四種 SuS 異構，在不連續膠體電泳之泳動率相異。異構之原態分子量約為 363,000, 次單元體分子量約為 90,000，推測其為四元體之構造。所有異構皆以 UDP 為其最佳核雙磷酸基質。所有異構蔗糖降解方向活性皆會被鎂離子抑制，而蔗糖合成方向活性則皆會受其誘導。

Both cytoplasmic and membrane-associated forms of sucrose synthase (SuS) were detected in the shoot of bamboo Lelebba oldhami. The content of membrane-associated SuS was low; but the specific activity was high. The interaction between membrane and SuS was weak, indicating SuS was a peripheral membrane protein. The cytoplasmic sucrose synthase was partially purified by ammonium sulfate fractionation, Sephacryl S-300 gel filtration and DEAE-Sephacel ion exchange chromatography. Further purification of the enzyme by the Mono-P chromatofocusing chromatography indicated the presence of at least four isozymes. These isozymes showed different electrophoretic mobilities in Disc-PAGE. The native and subunit molecular weight of these isozymes were 363,000 and 90,000, respectively. It suggests that SuS isozymes were tetramers. All these isozymes use UDP as the best nucleoside diphosphate substrate. The sucrose cleavage activities of these enzymes were inhibited by Mg2+ ion. However, activities of sucrose synthesis were stimulated by the same ion.

目錄
縮寫表 1
中文摘要 2
英文摘要 3
第一章 緒論 4
第一節 蔗糖合成之生化特性及生理意義 4
第二節 蔗糖合成異構之研究 6
第三節 本論文之緣起與研究目的 8
第二章 材料與方法 10
第一節 實驗材料 10
第二節 實驗儀器 10
第三節 實驗方法 11
第三章 結果與討論 21
第一節 竹筍膜結合型 SuS 21
第二節 竹筍可溶性 SuS 之純化 22
第三節 竹筍可溶性 SuS 生化性質之探討 24
第四節 竹筍不同部位 SuS 之比較 26
第四章 結論與展望 27
圖表 29
參考文獻 48

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. Proc. Natl. Acad. Sci. USA 92, 9353-9357.
Anguenot, R., Yelle, S. and Nguyen-Quoc, B. (1999). Purification of tomato sucrose synthase phosphorylated isoforms by Fe(III)-immobilized metal affinity chromatography. Arch. Biochem. Biophys. 365, 163-169.
Cardini, C.E., Leloir, L.F. and Chiriboga, J. (1995). The biosynthesis of sucrose. J. Biol. Chem. 214, 149-155.
Carlson, S.J. and Chourey, P.S. (1996). Evidence for plasma membrane-associated forms of sucrose synthase in maize. Mol. Gen. Genet. 252, 303-310.
Chan, H.Y., Ling, T.Y., Juang, R.H., Ting, I.N., Sung., H.Y. and Su, J.C. (1990) Sucrose synthase in rice plants. Plant Physiol. 94, 1456-1461.
Chopra, S., Del favero, J., Dolferus, R. and Jacobs, M. (1992). Sucrose synthase of Arabidopsis: genomic cloning and sequence characterization. Plant Mol. Biol. 18, 131-134.
Chourey, P.S. and Nelson, O.E. (1976). The enzymatic deficiency conditioned by the shrinken-1 mutation in maize. Biochem. Genet. 14, 1041-1055.
Chourey, P.S., Latham, M. and Still, P.E. (1986) Expression of two sucrose synthetase genes in endosperm and seedling cells of maize: evidence of tissue specific polymerization of promoters. Mol. Gen. Gent. 103, 251-255.
Chourey, P.S., Taliercio, E.W. and Kane, E.J. (1991). Tissue -specific expression and anaerobically induced post-transcripitional modulation of sucrose synthase gene in Sorghum bicolor M. Plant Physiol. 96, 485-490.
Cross, K.C. and Pharr, D.M. (1982). Cucumber fruit sucrose synthase isozymes. Phytochemistry 21, 1241-1244.
Echt, C. S. and Chourey, P. S. (1985) A comparison of two sucrose synthetase isozymes from normal and shrunken-1 maize. Plant Physiol. 79, 530-536.
Fu, H., Kim, S.Y. and Park, W.D. (1995). High-level tuber expression and sucrose inducibility of a potato Sus4 sucrose synthase gene require 5'' and 3'' flanking sequences and the leader intron. Plant Cell 7, 1387-1394.
Fu, H., Kim, S.Y. and Park, W.D. (1995). A potato Sus3 sucrose synthase gene contains a context-dependent 3'' element and a leader intron with both positive and negative tissue- specific effects. Plant Cell 7, 1395-1403.
Geigenberger, P. and Stitt, M. (1993). Sucrose synthase catalyses a readily reversible reaction in vivo iv developing potato tubers and other plant tissues. Planta 189, 329-339.
Huang, D.Y. and Wang, A.Y. (1998). Purification and characterization of sucrose synthase isozymes from etiolated rice seedlings. Biochem. Mol. Biol. Int. 46, 107-113.
Hisajima, S. and Ito, T. (1981). Purification and properties of sucrose synthase from morning-glory callus cells. Biol. Plant. 23, 356-364.
Hubbard, N.L., Pharr, D.M. and Huber, S.C. (1991). Sucrose phosphate synthase and acid invertase as determinants of sucrose concentration in developing muskmelon (Cucumis melo L.) fruits. Plant Physiol. 91, 1572-1574.
Koch, K. E., Notle, 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. (1996). Carbohydrate-modulated gene expression in plant. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47, 509-540.
Larsen, A.E., Salerno, G.L. and Points, H.G. (1985) Sucrose synthase from wheat leaves. Comparison with the weat germ enzyme. Physiol. Plant. 67:37-42.
McCarty, D.R., Shaw, J.R. and Hannah, L.C. (1986) The cloning, genetic mapping, and expression of the constitutive sucrose synthase locus of maize. Proc. Natl. Acad. Sci. USA. 83: 9099-9103.
Marana, C., Garcia-Olmedo, F. and Carbonero, P. (1988). Linked sucrose synthase genes in group-7 chromosomes in hexaploid wheat (Triticum aestivum L.). Gene 63, 253-260.
Marana, C., Garcia-Olmedo, F. and Carbonero, P. (1990). Differential expression of two types of sucrose synthase-encoding genes in wheat in response to anaerobiosis, cold shock and light. Gene 88, 167-172.
Moriguchi, T. and Ymaki, S. (1988). Purification and charaterization of sucrose synthase from peach (Prunus persci) fruit. Plant Cell Physiol. 29, 1361-1366.
Nasciment, R.O., Cordenunsi, R.B. and Lajolo, M.F. (1997). Partial purification and charaterization of Sucrose phosphate synthase form preclimateric and climacteric bananas. J. Agric. Food Chem. 45, 1103-1107.
Nguyen-Quoc, B., Krivitzky, M., Huber, S. C. and Lecharny, A. (1990). Sucrose synthase in developing maize leaves. Plant Physiol. 94, 516-523.
Pozueta-Romero, J., Yamaguchi, J., and Akazawa, T. (1991). ADPG formation by the ADP-specific cleavage of sucrose-reassessment of sucrose synthase. FEBS Lett. 291, 233-237.
Sanchez, d.l.H., Vicente-Carbajosa, J., Mena, M. and Carbonero, P. (1992). Homologous sucrose synthase genes in barley (Hordeum vulgare) are located in chromosomes 7H (syn. 1) and 2H. Evidence for a gene translocation? FEBS Lett. 310, 46-50.
Schaffer, A.A., Aloni. B. and Fogelman. E. (1987). Sucrose metabolism and accumulation in developing fruit of cucumis. Phytochemistry 26, 1883-1887.
Silvius, J.E. and Snyder, F.W. (1979). Comparative enzymic studies of sucrose metabolism in taproots and fibrous roots of Beta vulgaris L.. Plant Physiol. 64, 1070-1073.
Springer, B., Werr, W., Starlinger, P., Bennett, D.C., Zokolica, M., and Freeling, M. (1986). The Shrunken gene on chromosome 9 of Zea mays L. is expressed in various plant tissues and encodes an anaerobic protein. Mol. Gen. Genet. 205, 461-468.
Sturm, A. and Tang, G.Q. (1999). The sucrose-cleaving enzymes of plants are crucial for development, growth and carbon partitioning. Trends Plant Sci. 4, 401-407.
Sturm, A., Lienhard, S., Schatt, S. and Hardegger, M. (1999). Tissue-specific expression of two genes for sucrose synthase in carrot (Daucus carota L.). Plant Mol. Biol. 39, 349-360.
Su, J.C. (1965) Carbohydrate metabolism in the shoots of bamboo Leleba oldhaami. I Preliminary survey of soluble saccharides and sucrose-degrading enzymes. Bot. Bull. Acad. Sinica, 6, 153-159.
Su, J.C., Wu, J.L. and Yang, C.L. (1977). Purification and characterization of sucrose synthetase from the shoot of bamboo Leleba oldhanmi. Plant Physiol. 60, 17-21.
Su, J.C. and Preiss, J. (1978). Purification and properties of sucrose synthase from maize kernels. Plant Physiol. 61, 389-393.
Sung, H.Y., Yuan, H.F. and Su, J.C. (1971) Carbohydrate metabolism in the shoots of bamboo Leleba oldhaami. IX Some properties of the pentosan synthesizing system. J. Chin. Agric. Chem. Soc. Sp Iss, 62-73.
Taliercio, E.W. and Chourey, P.S. (1989). Post-transcripitional control of sucrose synthase expression on anaerobic seddlings of maize. Plant Physiol. 90, 1359-1364.
Tanase,K. and Yamaki,S. (2000). Purification and characterization of two sucrose synthase isoforms from Japanese pear fruit. Plant Cell Physiol. 41, 408-414.
Tupy, J. and Primot, L. (1982). Sucrose synthetase in the latex of Hevea Brasiliensis Muell. Arg.. J. Exp. Bot. 33, 988-995.
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 Mol. Biol. 18, 1191-1194.
Wang, F., Smith, A.G. and Brenner, M.L. (1993). Isolation and sequencing of tomato fruit sucrose synthase cDNA. Plant Physiol. 103, 1463-1464.
Weber, H., Borisjuk, L., Heim, U., Sauer, N. and Wobus, U. (1997). A role for suger transporters during seed development: molecular characterization of a hexose and a sucrose carrier in fava bean seed. Plant Cell 7, 1835-1846.
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 Lett. 420, 151-155.
Winter, H., Huber, J.L. and Huber, S.C. (1998). Identification of sucrose synthase as an actin-binding protein. FEBS Lett. 430 , 205-208.
Wolosiuk, R.A. and Pontis, H.G. (1971). Evidence of the existence of two forms of sucrose synthetase. FEBS Lett. 16, 237-240.
Yeh, D.B. and Su, J.C. (1972). Purification and charaterization of sucrose synthetase form banana fruits. J. Chin. Biochem. Soc. 1, 37-52.
Zeng, Y., Wu, Y., Avigne, W.T. and Koch, K.E. (1998). Differential regulation of sugar-sensitive sucrose synthases by hypoxia indicate complementary transcriptional and post transcriptional responses.
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 J. 7, 97-107.
王素秋 (1989) 豌豆蔗糖合成之研究, 碩士論文, 國立台灣大學農業化學系.
李根永(1976) 水稻穀實蔗糖合成酵素之性質, 碩士論文, 國立台灣大學農業化學系.
宋賢一 (1973) 蔗糖合成酵素在植物醣類代謝之功用, 博士論文, 國立台灣大學農業化學系.
岩素芬 (1992) 水稻蔗糖合成異構之純化與鑑定, 博士論文, 國立台灣大學農業化學系.
高閬仙 (1977) 豌豆芽中蔗糖合成酵素之動力學及反應機構之研究, 碩士論文, 國立台灣大學農業化學系.
張久瑗 (1978) 綠竹筍之蔗糖合成酵素之純化及第四級結構之探討, 碩士論文, 國立台灣大學農業化學系.
張毓秀 (1991) 台濃 67 號水稻葉片蔗糖合成之研究, 碩士論文, 國立台灣大學農業化學系.
黃德宜（1996）水稻白化苗蔗糖合成異構的純化及生化性質探討, 碩士論文, 國立台灣大學農業化學系.
程士瑜 (1987) 水稻蔗糖合成缺陷突變株及癒創組織蔗糖合成之免疫性質比較, 碩士論文, 國立台灣大學農業化學系.
鄭麗麗 (1974) 香蕉果實蔗糖合成酵素之動力學及反應機構之研究, 碩士論文, 國立台灣大學農業化學系.