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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李平篤(Ping-Du Lee) | |
dc.contributor.author | Chin-Lin Huang | en |
dc.contributor.author | 黃靜琳 | zh_TW |
dc.date.accessioned | 2021-06-15T01:26:07Z | - |
dc.date.available | 2010-07-27 | |
dc.date.copyright | 2009-07-27 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-23 | |
dc.identifier.citation | 參考文獻
Arai, T.; Hosoya, M.; Nakamura, M.; Magoori, E.; Uematsu, Y.; Sako, T. (2002) Cytosolic ratio of malate dehyrogenase/lactate dehydrogenase activity in peripheral leukocytes of race horses with training. Res Vet Sci , 72, (3), 241-4. Aranda, A.; Maugeri, D.; Uttaro, A. D.; Opperdoes, F.; Cazzulo, J. J.; Nowicki, C. (2006) The malate dehydrogenase isoforms from Trypanosoma brucei: subcellular localization and differential expression in bloodstream and procyclic forms. Int J Parasitol, 36, (3), 295-307. Banaszak L. J., Bradshow R. A. (1975) Malate dehydrogenase. In: The Enzymes, 3rd.ed (Ed. P. Boyer), 11, pp.369-396, Academic Press, New York Berkemeyer, M.; Scheibe, R.; Ocheretina, O. (1998) A novel, non-redox-regulated NAD-dependent malate dehydrogenase from chloroplasts of Arabidopsis thaliana L. J Biol Chem, 273, (43), 27927-33. Birktoft J. J., Bradshaw R. A., Banaszak L. J. (1987) Structure of porcine heart cytoplasmic malate dehydrogenase: combing X-ray diffraction and chemical sequence data in structural studies. Biochemistry USA 26, 2722-2734 Birkoft J. J., Fu Z.,Carnahan G. E, Rhodes G.,Roderick S L.,Banazak L. J. (1998) Comparsion of the molecular structure of cytoplasmic and mitochondrial malate dehydrogenase. Biochem. Soc. Trans. 17, 301-304 Birktoft J. J.. Rhodes G., Banaszak L. J. (1989) Refined crystal structure of cytoplasmic malate dehydrogenase at 2.5 A resolution. Biochemistry USA 28, 6065-6089 Cherniad'ev, II, (2002) Effect of preparations exhibiting cytokinin-like activity on the specific density of leaf in grasses. Prikl Biokhim Mikrobiol , 38, (6), 689-97. Chen, J.; Smith, D. L. (2001) Amide hydrogen exchange shows that malate dehydrogenase is a folded monomer at pH 5. Protein Sci, 10, (5), 1079-83. Clarke, A. R.; Wigley, D. B.; Chia, W. N.; Barstow, D.; Atkinson, T.; Holbrook, J. J. (1986) Site-directed mutagenesis reveals role of mobile arginine residue in lactate dehydrogenase catalysis. Nature, 324, (6098), 699-702 Cox, B.; Chit, M. M.; Weaver, T.; Gietl, C.; Bailey, J.; Bell, E.; Banaszak, L. (2005) Organelle and translocatable forms of glyoxysomal malate dehydrogenase. The effect of the N-terminal presequence. Febs J , 272, (3), 643-54. David L. Nelson, M. M. C. (2005) Lehninger PRINCIPLES OF BIOCHEMISTRY. 4 ed.; W. H. Freeman and Company. Faske, M.; Backhausen, J. E.; Sendker, M.; Singer-Bayrle, M.; Scheibe, R.; Von Schaewen, A. (1997) Transgenic Tobacco Plants Expressing Pea Chloroplast Nmdh cDNA in Sense and Antisense Orientation (Effects on NADP-Malate Dehydrogenase Level, Stability of Transformants, and Plant Growth). Plant Physiol , 115, (2), 705-715. Feeney, R.; Clarke, A. R.; Holbrook, J. J. (1990) A single amino acid substitution in lactate dehydrogenase improves the catalytic efficiency with an alternative coenzyme. Biochem Biophys Res Commun, 166, (2), 667-72. Gelpi, J. L.; Dordal, A.; Montserrat, J.; Mazo, A.; Cortes, A. (1992) Kinetic studies of the regulation of mitochondrial malate dehydrogenase by citrate. Biochem J , 283 ( Pt 1), 289-97 Genda, T.; Nakamatsu, T.; Ozak, H. (2003) Purification and characterization of malate dehydrogenase from Corynebacterium glutamicum. J Biosci Bioeng, 95, (6), 562-6. Gietl C. (1992) Malate dehydrogenase isoenzymes: cellular locations and role in the flow of metabolites between the cytoplasm and cell organelles. Biochem. Biophys. Acta1100, 217-234 Gietl, C.; Hock, B. (1987) Glyoxysomal and mitochondrial malate dehydrogenase in watermelon cotyledons. Isozymes Curr Top Biol Med Res, 16, 175-92. Gietl, C.; Lehnerer, M.; Olsen, O. (1990) Mitochondrial malate dehydrogenase from watermelon: sequence of cDNA clones and primary structure of the higher-plant precursor protein. Plant Mol Biol , 14, (6), 1019-30. Gietl, C. (1992) Partitioning of Malate Dehydrogenase Isoenzymes into Glyoxysomes, Mitochondria, and Chloroplasts. Plant Physiol, 100, (2), 557-559. Goward C. R., Nicholls D. J. (1994) Malate dehydrogenease: a model for structure, evolution and catalysis. Protein Sci. 3, 1883-1888 Hartl F. U., Pfanner N., Nicholson D., Neupert W. (1989) Mitochondril protein import. Biochim. Biophys. Acta 988 1-45 Hatch M. D., Slack C. R. (1969) NAD-specific malate dehydrogenase and glycerate kinase in leaves and evidence for their location in chloroplasts. Biochem. Biophys. Res. Commun. 34, 589-593 Hayes, M. K.; Luethy, M. H.; Elthon, T. E. (1991) Mitochondrial Malate Dehydrogenase from Corn : Purification of Multiple Forms. Plant Physiol, 97, (4), 1381-1387. Hecht K., Wrba A., Jaenicke R. (1989) Catalytic properties of thermophilic lactate dehydrogenase and halophilic malate dehydrogenase at high temperature and low water activity. Eur. J. Biochem. 183, 69-74 Hock B., Gietl C. (1982) Cell-free synthesis of water-melon glyoxysomal malate dehydrogenase: a comparison with the mitochondrial isoenzyme. Ann. NY Acad Sci. 386, 350-361 Hung, H. C.; Kuo, M. W.; Chang, G. G.; Liu, G. Y. (2005) Characterization of the functional role of allosteric site residue Asp102 in the regulatory mechanism of human mitochondrial NAD(P)+-dependent malate dehydrogenase (malic enzyme). Biochem J, 392, (Pt 1), 39-45. Issakidis, E.; Saarinen, M.; Decottignies, P.; Jacquot, J. P.; Cretin, C.; Gadal, P.; Miginiac-Maslow, M. (1994) Identification and characterization of the second regulatory disulfide bridge of recombinant sorghum leaf NADP-malate dehydrogenase. J Biol Chem, 269, (5), 3511-7. Joh T., Takeshima H., Tsuzuki T., Setoyama C., Shimada K., Tanase S., Kuramitsu S., Kagamiyama H., Morino Y. (1987) Cloning and sequence analysis of cDNAs encoding mammalian cytosolic malate dehydrogenase. Comparison of the amino acid sequences of mammalian and bacterial malate dehydrogenase. J. Biol. Chem. 262, 15127-15131 Johnson H. P., Hatch M. D. (1970) Properties and regulation of leaf nicotinamide-adenine dinucleotide phosphate-malate dehydrogenase and “malic” enzyme in plants with the C4-dicarboxylic acid pathway of photosynthesis. Biochem. J. 119. 273-280 Kim, D. J.; Smith, S. M. (1994) Expression of a single gene encoding microbody NAD-malate dehydrogenase during glyoxysome and peroxisome development in cucumber. Plant Mol Biol , 26, (6), 1833-41. Leroux, A.; Fleming-Canepa, X.; Aranda, A.; Maugeri, D.; Cazzulo, J. J.; Sanchez, M. A.; Nowicki, C. (2006) Functional characterization and subcellular localization of the three malate dehydrogenase isozymes in Leishmania spp. Mol Biochem Parasitol , 149, (1), 74-85. Miller, C. O. (1980) Cytokinin inhibition of respiration in mitochondria from six plant species. Proc Natl Acad Sci U S A, 77, (8), 4731-4735. Madern, D.; Cai, X.; Abrahamsen, M. S.; Zhu, G. (2004) Evolution of Cryptosporidium parvum lactate dehydrogenase from malate dehydrogenase by a very recent event of gene duplication. Mol Biol Evol, 21, (3), 489-97. Magori, E.; Nakamura, M.; Inoue, A.; Tanaka, A.; Sasaki, N.; Fukuda, H.; Mizutani, H.; Sako, T.; Kimura, N.; Arai, T. (2005) Malate dehydrogenase activities are lower in some types of peripheral leucocytes of dogs and cats with type 1 diabetes mellitus. Res Vet Sci , 78, (1), 39-44. Nishiyama, M.; Birktoft, J. J.; Beppu, T. (1993) Alteration of coenzyme specificity of malate dehydrogenase from Thermus flavus by site-directed mutagenesis. J Biol Chem, 268, (7), 4656-60. Parker, D. M.; Lodola, A.; Holbrook, J. J. (1978) Use of the sulphite adduct of nicotinamide-adenine dehydrogenase. Biochem J , 173, (3), 959-67. Portilla-Arias, J. A.; Garcia-Alvarez, M.; Galbis, J. A.; Munoz-Guerra, S. (2008) Biodegradable Nanoparticles of Partially Methylated Fungal Poly(beta-L-malic acid) as a Novel Protein Delivery Carrier. Macromol Biosci. Piera-Velazquez, S.; Marhuenda-Egea, F.; Cadenas, E. (2002) Increased stability of malate dehydrogenase from Halobacterium salinarum at low salt concentration in reverse micelles. Extremophiles , 6, (5), 407-12. Parker, D. M.; Lodola, A.; Holbrook, J. J. (1978) Use of the sulphite adduct of nicotinamide-adenine dehydrogenase. Biochem J , 173, (3), 959-67. R. A. Musrati, M. Kollarova, N. Mernik and D. Mikulasova. (1998) Malate Dehydrogenase : Distribution, Function and Properties. Gen.Physiol. Biophys., 17, 193-210 Randokph T Wedding, M. Kay Black and Dennis Pap. (1976) Malate Dehydrogenase and NAD Malic Enzyme in the Oxidation of Malate by Sweet Potato Mitochondria. Plant physiol. 58, 740-743 Ruelland, E.; Lemaire-Chamley, M.; Le Marechal, P.; Issakidis-Bourguet, E.; Djukic, N.; Miginiac-Maslow, M. (1997) An internal cysteine is involved in the thioredoxin-dependent activation of sorghum leaf NADP-malate dehydrogenase. J Biol Chem , 272, (32), 19851-7. Savenstrand, H.; Strid, A. (2004) A Pisum sativum glyoxysomal malate dehydrogenase induced by cadmium exposure. DNA Seq, 15, (3), 206-8. Savitch, L. V.; Barker-Astrom, J.; Ivanov, A. G.; Hurry, V.; Oquist, G.; Huner, N. P.; Gardestrom, P. (2001) Cold acclimation of Arabidopsis thaliana results in incomplete recovery of photosynthetic capacity, associated with an increased reduction of the chloroplast stroma. Planta , 214, (2), 295-303. Scheibe, R. (2004) Malate valves to balance cellular energy supply. Physiol Plant, 120, (1), 21-26. Setoyama C., Ding S. H., Choudhury B. K., Joh T., Takeshima H., Tsuzuki T., Shimada K. (1990) Regulatory regions of the mitochondrial and cytosolic isoenzyme genes participating I the malate-aspartate shuttle. J. Biol. Chem. 265, 1293-1299 Strumilo, S.; Ovseniuk, A.; Radeczka, A.; Tylicky, A. (2006) [Comparison of malate dehydrogenase I sozymes from the hare and rabbit heart]. Zh Evol Biokhim Fiziol, 42, (5), 450-2. Tomita, T.; Fushinobu, S.; Kuzuyama, T.; Nishiyama, M. (2005) Crystal structure of NAD-dependent malate dehydrogenase complexed with NADP(H). Biochem Biophys Res Commun, 334, (2), 613-8 Wilks H. M., Hart K. N., Feeney R., Dunn C. R., Murhead H., Chia W. N., BarstowD. A., Atkinson T., Clarke A. R., Holbrook J. J. (1988) A specific, highlyactive malate dehydrogenase by redesign of a lactate dehydrogenase framework.Science 242, 1541-1544 Yueh A. Y., Chung C.-S., Lai Y.-K. (1989) Purification and molecular properties of malate dehydrogenase from the marine diatom Nitzschia alba. J. Biochem. 258, 221-228 謝陸盛 (2005) 博士論文初稿,國立臺灣大學微生物與生化學研究所 蕭哲仁 (2008) 碩士論文,國立臺灣大學微生物與生化學研究所 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/42854 | - |
dc.description.abstract | 蘋果酸去氫酶 (malate dehydrogenase, MDH EC 1.1.1.37) 廣泛存在生物體中。負責可逆性催化oxaloacetate 和malate間的轉換。蘋果酸去氫酶在高等植物中有多種異構酶,存在不同的胞器中,使用不同的輔酶,對於催化之生理途徑有其專一性。
本實驗室之前於綠竹cDNA庫篩選細胞分裂素氧化酶/去氫酶時,意外篩到了一可能為綠竹gMDH的基因,因此對MDH產生極大興趣,本實驗進行甘藷MDH之純化、生化性質及酵素動力學研究。 從甘藷中純化MDH,經緩衝液粗抽、硫酸銨分劃、膠體過濾法 (Sephacryl S-300) 及親和層析法 (Blue Sepharose¬¬¬ CL-4B) 得到的純化蛋白質,其反應最適反應溫度為45 ℃,最適pH值為10~10.5;金屬離子會提高MDH活性;重金屬離子以Hg2+抑制MDH活性最甚;而化學修飾劑DEPC可抑制MDH活性,推測MDH之活性區可能含有histidine;MDH所參與之生理途徑之下游產物皆對MDH有抑制作用。 在酵素動力學中,MDH對malate之Vmax值為1.12 nmol / min,Km值為0.21 mM;對NAD+之Vmax值為4.41 nmol/min,Km值為0.18 mM。從膠體過濾法及原態膠體電泳可推測MDH原態分子量約為70 kD;從SDS-PAGE得知MDH單體約為34 kD,推測甘藷MDH應為同質二元體。 | zh_TW |
dc.description.abstract | Malate dehydrogenase (MDH EC 1.1.1.37) is a ubiquitous enzyme in all kinds of living organisms. MDH catalyzes the interconversion of oxaloacetate and malate. Higher plants contain multiple forms of MDH that differ in co-enzyme specificity, subcellular localization and physiological function.
Unexpectedly, a putative mdh cDNA was screened with the specific probe of cytokinin from the cDNA library of Bambusa oldhamii in our laboratory. For comparing the MDH of bamboo, we continue studying about MDH in sweet potato. We purified MDH from sweet potato through buffer extraction, ammonium sulfate precipitation, gel filtration (Sephacryl S-300) and affinity chromatography (Blue Sepharose CL-4B). The optimal temperature of MDH is 45 ℃; the optimal pH is 10-10.5; MDH activity was benefited by metal ions, but heavy metal ions inhibited its activity. DEPC inhibited MDH activity through modifying histidine. About the MDH enzyme kinetics, the Vmax of malate is 1.12 nmol / min and the Km is 0.21 mM; the Vmax of NAD+ is 4.41 nmol / min and the Km is 0.18 mM. The molecular weight of native MDH was estimated to be 70 kD and the subunit of MDH was about 34 kD. It is estimated that MDH might be a homodimeric enzyme. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T01:26:07Z (GMT). No. of bitstreams: 1 ntu-98-R96B47212-1.pdf: 3901856 bytes, checksum: 7a9766feb031e8fa25d5c589f211fe5c (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 目錄
縮寫表 IV 摘要 V Abstract VI 第一章 緒論 1 第一節 實驗緣起 1 第二節 蘋果酸 2 第三節 蘋果酸所參與之生理途徑 3 第四節 蘋果酸調控之相關酵素 9 第五節 蘋果酸去氫酶 10 第六節 蘋果酸去氫酶異構酶之研究 11 第七節 蘋果酸去氫酶序列之演化研究 15 第八節 蘋果酸去氫酶蛋白質結構與作用機制 17 第九節 蘋果酸去氫酶與CKX的相關研究 18 第二章 材料與方法 20 第一節 實驗材料、藥品與儀器 20 2.1.1 實驗材料 20 2.1.2 實驗藥品與儀器 20 第二節 一般實驗法 21 2.2.1 Malate Dehydrogenase酵素活性測定 21 2.2.2 蛋白質定量法Bradford Method 23 2.2.3 蛋白質電泳檢定系統 25 2.2.3.1 原態膠體電泳 25 2.2.3.2 蛋白質染色法Coomassie Brilliant R-250 27 2.2.3.3 SDS膠體電泳 28 2.2.3.4 膠片乾燥法 30 2.2.3.5 蛋白質轉印法 31 2.2.3.6 免疫染色法 32 第三節 甘藷Malate Dehydrogenase之純化 34 2.3.1 粗抽取及硫酸銨分劃 34 2.3.2 膠體過濾法 Sephacryl S-300 36 2.3.3 親和層析法 Blue Sepharose CL-4B 37 2.3.4 快速蛋白質液相層析 Fast Protein Liquid Chromatography 38 2.3.5 Mono Q HR5/5 chromatography 39 第四節 甘藷Malate Dehydrogenase生化性質探討 41 2.4.1 最適反應條件 41 2.4.2 最適反應溫度 41 2.4.3 活化能測定 42 2.4.4 最適反應pH 42 2.4.5 金屬離子對活性影響 44 2.4.6 重金屬離子對活性影響 44 2.4.7 化學修飾劑對活性影響 45 2.4.8 MDH參與之生理途徑下游產物對活性影響 46 2.4.9 MDH動力學分析 47 2.4.9.1 MDH對malate結合能力 47 2.4.9.2 MDH對NAD+結合能力 48 第三章 實驗結果與討論 49 第一節 甘藷Malate Dehydrogenase之純化 49 3.1.1 純化步驟 49 3.1.2 粗抽取及硫酸銨分劃 49 3.1.3 膠體過濾法 Sephacryl S-300 50 3.1.4 親和層析法 Blue Sepharose CL-4B 50 3.1.5 原態分子量測定 51 第二節 甘藷Malate Dehydrogenase生化性質探討 52 3.2.1 最適反應溫度 52 3.2.2 反應活化能 (Ea) 52 3.2.3 最適反應pH 52 3.2.4 金屬離子對MDH活性影響 53 3.2.5 重金屬離子對MDH活性影響 53 3.2.6 化學修飾劑對MDH活性影響 54 3.2.7 MDH參與之生理途徑下游產物對MDH活性影響 55 3.2.8 MDH之酵素動力學分析 56 第四章 結論與展望 57 第一節 甘藷Malate Dehydrogenase之純化方法 57 第二節 甘藷Malate Dehydrogenase之生化性質 57 第三節 未來展望 58 結果圖表集 60 附錄 82 參考文獻 86 問答集 90 | |
dc.language.iso | zh-TW | |
dc.title | 甘藷蘋果酸去氫酶之生化學研究 | zh_TW |
dc.title | Biochemical Studies of Malate Dehydrogenase
from Sweet Potato (Ipomoea batatas) | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 林棋財,楊健志 | |
dc.subject.keyword | 蘋果酸去氫酶,同質二元體, | zh_TW |
dc.subject.keyword | malate dehydrogenase,homodimer, | en |
dc.relation.page | 90 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-07-23 | |
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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