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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李平篤(Ping-Du Lee) | |
| dc.contributor.author | Pei-Jou Chu | en |
| dc.contributor.author | 朱珮柔 | zh_TW |
| dc.date.accessioned | 2021-06-13T02:17:46Z | - |
| dc.date.available | 2007-02-27 | |
| dc.date.copyright | 2007-02-27 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-02-02 | |
| dc.identifier.citation | Ann YG, Iizuka M, Minamura N (1989) Evidence for existence of an active monomer of sweet potato β-amylase. Agric. Biol. Chem. 53(11): 3109-3110
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Plant Foods for Human Nutrition 58: 309–315 Lao NT, Schoneveld O, Mould RM, Hibberd JM, Gray JC, Kavanagh TA (1999) An Arabidopsis gene encoding a chloroplast-targeted beta-amylase. Plant J. 20: 519-527 Lee EYC (1971) The action of sweet potato β-amylase on glycogen and amylopectin: Formation of a novel limit dextrin. Arch. Biochem. Biophys. 240(2): 757-767 Lizotte PA, Henson, Duke SH (1990) Purification and characterization of pea epicotyl β-amylase. Plant Physiol. 92: 615-621 Monroe JD, Preiss J (1990) Purification of a β-amylase that accumulates in Arabidopsis thailand mutants defective in starch metabolism. Plant Physiol. 94: 1033-1039 Nakamura K, Ohto M, Yoshida N (1991) Sucrose-induced accumulation of β-amylase occurs concomitant with the accumulation of starch and sporamin in leaf-petiole cuttings of sweet potato. Plant Physiol. 96: 902-909 Nielsen T, Deiting U, Stitt M (1997) A β-amylase in potato tubers is induced by storage at low temperature. Plant Physiol 113: 503-510 Robyt JF (1984) Enzymes in the hydrolysis and synthesis of starch. In “Starch” 2nd ed, Whistler RL, Bemiller JN, Paschall EF (Eds.) Academic Press Inc, New York, pp.87-123 Sawai J, Nakai T, Hashimoto A, Shimizu M (2004) A comparison of the hydrolysis of sweet potato starch with β-amylase and infrared radiation allows prediction of reducing sugar production. International Journal of Food Science and Technology 39: 967-974 Sharma A, Sharma S, Gupta MN (2000) Purification of wheat germ amylase by precipitation. Protein Expression and Purification 18: 111-114 Sharma CB, Goel M, Irshad M (1978) Myoinositol hexaphosphate as a potential inhibitor of α-amylases. Phytochemistry 17: 201-204 Teotia S, Gupta MN (2001) Reversibly soluble macroaffinity ligand in aqueous two-phase separation of enzymes. J Chromatogr A 923(1-2): 275-80 Teotia S, Gupta MN (2002) Magnetite-alginate beads for purification of some starch degrading enzymes. Mol Biotechnol 20(3): 231-7 Teotia S, Khare SK, Gupta MN (2001) An efficient purification process for sweet potato beta-amylase by affinity precipitation with alginate. Enzyme and Microbial Technology 28: 792-795 Thoma JA, Spradlin JE, Dygert S (1971) Plant and animal amylase. In PD Boyer, ed, The enzyme, Ed.3, Academic press, New York, Vol 5. 115-189 Thoma JA, Wakim J, Stewart L (1963) Comparison of the active sites of alpha and beta amylases. Biochem. Biophys. Res. Commun. 12: 350-355 Wang Q, Monroe J, Sjolund RD (1995) Identification and characterization of a phloem-specific β-amylase. Plant Physiol 109: 743-750 Wang Q, Monroe J, Sjolund RD (1995) Identification and characterization of a phloem-specific beta-amylase. Plant Physiol. 109: 743-750 Wang SM, Liu WL, Eimert K, Chen J (1996) Phytohormone-regulated β-amylase genes expression in rice. Plant Mol. Biol. 31: 975-982 Yamamoto T (1995)-b, In: The amylase research society of Japan (ed) Enzyme chemistry and molecular biology of amylases and related enzymes. Yamasaki Y (2003) β-amylase in germinating millet seeds. Phytochemistry 64: 935–939 Young GH, Chen HM, Lin CT, Tseng KC, Wu JS, Juang RH (2006) Site-specific phosphorylation of L-form starch phosphorylase by the protein kinase activity from sweet potato roots. Planta 223: 468–478 Ziegler P, Beck E (1986) Exoamylase activity in vacuoles isolated from pea and wheat leaf protoplasts. Plant Physiol 82: 1119-1121 莊榮輝 (1985) 水稻蔗糖合成酶之研究。博士論文，國立台灣大學農業化學研究所 張瓈文 (1992) 生澱粉水解酶之研究（二）純化與生化性質再探討。碩士論文，國立中興大學食品科學研究所 鄒純安 (1993) 酵素法由甘藷製備高麥芽糖糖漿與副產品。碩士論文，國立台灣海洋大學水產食品科學研究所 陳師瑩 (1997) β-澱粉酶阻礙澱粉磷解酶的分子機轉。博士論文，國立台灣大學農業化學研究所 劉志寧 (1998) 甘藷癒創組織光照誘導β-澱粉酶之表現及其生化性質探討。碩士論文，國立台灣大學農業化學研究所 陳韋琮 (1999) 甘藷β-澱粉酶在大腸桿菌之表現及以原位雜交法探討其在甘藷塊根內之分布。碩士論文，國立台灣大學農業化學研究所 張弘儒 (2003) β-澱粉酶基質結合與催化機制的探討。碩士論文，國立台灣大學農業化學研究所 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30841 | - |
| dc.description.abstract | β-澱粉酶(β-amylase, EC 3.2.1.2，簡稱BA) 催化水解澱粉鏈之非還原端
α-1,4-糖甙鍵，釋出β-maltose而留下β-limit dextrin，分類上屬於外切澱粉酶。 本論文針對不同品種之甘藷測定BA活性，選擇總活性最高之60℃樣本與對照組進行純化，測定生化性質加以比較，來探討溫度對BA活性之影響。純化將粗抽液經硫酸銨100%沈澱後，進行製備式電泳可得到相當純的BA。 對照組與60℃樣本對基質澱粉之 Km 值分別為 6.02 與 2.95%。酵素最適反應溫度均為60℃，對照組最適反應pH為 5，60℃樣本為 4.5。對照組與 60℃樣本之活化能分別為 3.35 與 4.2 kcal/mol；酵素活性會受到汞、銅、銀離子的抑制，除了汞離子外，60℃樣本對金屬離子的耐受性均較對照組差。化學修飾劑β-mercaptoethanol、DTT、PMSF中除了DTT外，60℃樣本受到的影響較對照組來得大。 | zh_TW |
| dc.description.abstract | β-amylase (EC 3.2.1.2) is an exoamylase that releases successive maltose units from the nonreducing end of a polysaccharide chain by hydrolyzing of α-1,4-glucan linkages.
In this article, BA activities were measured in different varieties of sweet potato, and chose the highest total activity sample under 60℃ treatment and control sample to purify. And put to compare their biochemical characteristics and the effects of temperature on BA activity. Crude extraction precipitated by ammonium sulfate 100% then run preparative electrophoresis can get pure enzyme. Km values for starch substrate of control sample and 60℃ sample were estimated to be 6.02 and 2.95%, respectively. Optimal temperature for both samples are 60℃. Optimal pH value for control sample is 5, and for 60℃ sample is 4.5. Activation energy for control sample and 60℃ sample is 3.35 and 4.2 kcal/mol, respectively. BA activity could be inhibited by Hg2+, Cu2+ and Ag+. Except for Hg2+, tolerance to metal ions of 60℃ sample is less than control sample. Chemical modified reagents, including β-mercaptoethanol, DTT, PMSF also influence BA activity. Except for DTT, influence of chemical modified reagents of 60℃ sample is more than control sample. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T02:17:46Z (GMT). No. of bitstreams: 1 ntu-96-R93b47211-1.pdf: 917075 bytes, checksum: dce53647d0d72876289452e2c20b6bff (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 目錄…………………………………………………………………………...………..Ⅰ
縮寫表……………………………………..…………………………………………...III 中文摘要…………………………...………………………….………………...……..IV Abstract……...……..…………………………………………..………………………..V 第一章 緒論………………………………………………………………………...…1 1.1 甘藷…………………………………………………………………………....1 1.2 澱粉酶 (Amylase)…………………………………..………………………...2 1.2.1 α-amylase (AA) ………………….…………………...…………………3 1.2.2 β-amylase (BA) ………………….…………………...………….……...3 1.3 實驗緣起………………………………………………………………….…...9 第二章 材料與方法………………………………………………………………….10 2.1 實驗材料…………………………………………………………………......10 2.1.1 甘藷…………………..……………………………………………......10 2.1.2 馬鈴薯………………..……………………………………………......10 2.2 實驗藥品………………………………...…………………………………...10 2.3 儀器設備…………………………………………………………………......10 2.4 蛋白質定量法………………………………………………………..............11 2.5 β-澱粉酶活性分析法…………………………………………..……...13 2.5.1 活性分析………………………………………………………..13 2.5.2 澱粉酶活性染色………………………………………………..14 2.6 迷你電泳檢定系統…………………………………………..................15 2.6.1 原態膠體電泳………………………………………….…..…...16 2.6.2 SDS 膠體電泳…………………………………………….……19 2.7 膠體染色法………………………………………………………...…...22 2.7.1 Coomassie Brilliant Blue R-250 (CBR) 染色法………..….…..22 2.7.2 醣蛋白質染色法 (過碘酸-硝酸銀法)…………….….….…….23 2.7.3 膠片乾燥法…………………………….….……………………25 2.8 β-澱粉酶的純化…………………………….……….………………...26 2.8.1 粗抽及硫酸銨分劃…………………………………….……….26 2.8.2 製備式電泳與電泳溶離………………………………..………27 2.9 酵素生化性質…………………...……………….………………..……29 2.9.1 酵素反應最適 pH 值………………………………...…………29 2.9.2 酵素酸鹼安定性........………………….……………...…………29 2.9.3 最適反應溫度…………………...………………………….……30 2.9.4 活化能 (Activation energy, Ea) …………………...………….…30 2.9.5 熱安定性………………………………….…………...…………30 2.9.6 Km 值………………….................................................….………30 2.9.7 金屬離子對酵素活性之影響………………………...………….30 2.9.8 化學修飾物對酵素活性之影響…………………………...…….30 第三章 實驗結果…………………………………………………………………….32 3.1 溫度對β-澱粉酶活性之影響……….……………………………………….32 3.1.1 實驗流程…………………………………………...………………...32 3.1.2 溫度對不同品種甘藷 BA 之影響…….…………..………….….....32 3.1.3 甘藷與馬鈴薯中BA含量之對照…………………………….……..33 3.1.4 不同升溫處理方式對甘藷BA活性之影響………………….……..34 3.2 β-澱粉酶之純化………………………………………………...…….….....34 3.2.1 純化流程………………………….…………………….....................34 3.2.2 膠片染色結果………………………………………………....……..36 3.3 溫度對β-澱粉酶生化性質之影響…….................………………….……...37 3.3.1 酵素反應最適 pH 值………………………………...….……...37 3.3.2 酵素酸鹼安定性........………………….……………...….……...37 3.3.3 最適反應溫度…………………...………………………..……...37 3.3.4 活化能………………………….…………………...…….……...37 3.3.5 熱安定性………………………………….…………...…….…...39 3.3.6 酵素動力學研究...…….................................................…….…...39 3.3.7 金屬離子對酵素活性之影響………………………...……….…40 3.3.8 化學修飾物對酵素活性之影響…….…………………...………40 第四章 討論與展望………………………………………………………………….42 4.1 溫度對β-澱粉酶活性之影響………………………………………..….......42 4.2 β-澱粉酶之純化……………………………………………….....................43 4.3 溫度對β-澱粉酶生化性質之影響…...…………………………………......44 4.4 未來展望…….………………………………….............................................45 結果圖表集…………………………………………………………………………….47 參考文獻……………………………………………………………………………….69 | |
| dc.language.iso | zh-TW | |
| dc.subject | β-澱粉酶 | zh_TW |
| dc.subject | 溫度 | zh_TW |
| dc.subject | 甘藷 | zh_TW |
| dc.subject | temperature | en |
| dc.subject | β-amylase | en |
| dc.subject | sweet potato | en |
| dc.title | 溫度對甘藷β-澱粉酶活性影響之研究 | zh_TW |
| dc.title | Effect of temperature on β-amylase activity in sweet potato | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林棋財(Chi-Tsai Lin),楊健志 | |
| dc.subject.keyword | 溫度,甘藷,β-澱粉酶, | zh_TW |
| dc.subject.keyword | temperature,sweet potato,β-amylase, | en |
| dc.relation.page | 72 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-02-04 | |
| 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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