Studies on the Novel Enzyme Phenylalanine Dehydrogenase.
新型酶苯丙氨酸脱氢酶的研究。
基本信息
- 批准号:05044138
- 负责人:
- 金额:$ 2.56万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for international Scientific Research
- 财政年份:1993
- 资助国家:日本
- 起止时间:1993 至 1994
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
We discovered for the first time that Bacillus sp.and Sporosarcina sp.produces phenylalanine dehydrogenase. We started our collaboration with Professor Arthur Copper of Cornell University Medical College, New York, U.S.A.We got the following results in this two years collaboration with him.(1)We have clarified the enzymological properties of phenylalanine dehydrogenase from Bacillus shaericus, B.badius, and Sporosarcina ureae. We synthesized various optically pure L-amino acids from prochiral alpha-keto acids, using the enzyme in combination with Candida formate dehydrogenase. The enzyme was effective for the reductive amination of various alpha-Keto acids. We have been successful in the synthesis of natural and unnatural L-amino acids, including L-homophenylalanine.(2)We have been successful in the synthesis of ^<13>N-Labelled L-phenylalanine, L-tyrosine, and L-DOPA.(3)We have cloned the genes for the enzymes and sequenced some of the genes. They showed similarities with leucine dehydrogenase, Streptomyces phenylalanine dehydrogenase, and glutamic acid dehydrogenase.(4)In the microplate method of PKU screening using the enzyme, L-phe is oxidized with the enzyme to phenylpyruvate, reducing NAD^+ to NADH.In the presence of NADH and the electron carrier 1-methoxy-5-methylphenazium methylsulfate (1-PMS), Co^<3+> is reduced to Co^<2+> which produces a colored chelate. The method is being introduced as a tool of the routine mass screening in Japan.(5)A spirolactone [3-hydroxy-2-oxo-1-oxaspiro [4.5] -dec-3-ene] and the sodium salts of two new alpha-keto acids [(1-cyclohexenyl) glyoxylate, beta- (1-hydoxycyclohexane) pyruvate (enol form)] were chemically synthesized. These compounds are expected to be useful as prototypes for the design of inhibitors of alpha-keto acid-utilizing enzymes. In fact, one of them was found to be a new type of inhibitor of rabbit muscle lactate dehydrogenase.
首次发现芽孢杆菌和孢子八叠球菌产生苯丙氨酸脱氢酶。我们与美国纽约康奈尔大学医学院的亚瑟科珀教授开始了合作。(1)We已经阐明了来自Bacillus shaericus、B.badius和Sporosarcina uericum的苯丙氨酸脱氢酶的酶学性质。我们合成了各种光学纯的L-氨基酸的前手性α-酮酸,使用的酶与假丝酵母甲酸脱氢酶的组合。该酶对各种α-酮酸的还原胺化是有效的。我们已经成功地合成了天然和非天然的L-氨基酸,包括L-高苯丙氨酸。(2)We已成功合成^<13>N-标记的L-苯丙氨酸、L-酪氨酸和L-DOPA。(3)We已经克隆了这些酶的基因,并对其中一些基因进行了测序。它们与亮氨酸脱氢酶、链霉菌苯丙氨酸脱氢酶和谷氨酸脱氢酶表现出相似性。(4)In在使用该酶进行PKU筛选的微孔板法中,L-苯丙氨酸被该酶氧化为苯丙酮酸,NAD^+被还原为NADH。在NADH和电子载体1-甲氧基-5-甲基吩嗪甲基硫酸盐(1-PMS)存在下,Co^<3+>被还原为Co^<2+>,生成有色螯合物。该方法正在日本作为常规大规模筛查的工具引入。(5)A化学合成了螺内酯[3-羟基-2-氧代-1-氧杂螺[4.5] -癸-3-烯]和两种新的α-酮酸[(1-环己烯基)乙醛酸、β-(1-羟基环己烷)丙酮酸(烯醇形式)]的钠盐。预期这些化合物可用作设计α-酮酸利用酶的抑制剂的原型。事实上,其中一种被发现是一种新型的兔肌肉乳酸脱氢酶抑制剂。
项目成果
期刊论文数量(32)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Abraham DG,Patel P,and Cooper AJL.: "Characterization of a high molecular weight cysteine-S-conjugate beta-lyase from rat kidney with activity toward leukotriene E4." Journal of Biological Chemistry. 270. 180-188 (1995)
Abraham DG、Patel P 和 Cooper AJL.:“大鼠肾脏中具有白三烯 E4 活性的高分子量半胱氨酸-S-缀合物 β-裂合酶的表征。”
- DOI:
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- 影响因子:0
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Makar TK,Cooper AJL,Tofel-Orehi B,Thaler TH,and Blass JP.: "Carnitine, carnitine acetyl translerase, and glutathione in Alzheimer brain." Neurochemical Research. (in press). (1995)
Makar TK、Cooper AJL、Tofel-Orehi B、Thaler TH 和 Blass JP.:“阿尔茨海默病大脑中的肉碱、肉碱乙酰转移酶和谷胱甘肽。”
- DOI:
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- 影响因子:0
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M.Ueda, H.Yamada and Y.Asano: "The Substrate Specificity of Maleate Hydratase from Arthrobacter sp. strain MCI2612." Applied Microbiology and Biotechnology. 41. 215-218 (1994)
M.Ueda、H.Yamada 和 Y.Asano:“来自节杆菌菌株 MCI2612 的马来酸水合酶的底物特异性。”
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- 影响因子:0
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Y.Asano, Y.Yamamoto, and H.Yamada: "Catechol-2,3-Dioxygenase-catalyzed Synthesis of Picolinic Acids from Catechols." Bioscience Biotechnology and Biochemistry. 58. 2054-2056 (1994)
Y.Asano、Y.Yamamoto 和 H.Yamada:“儿茶酚 2,3-双加氧酶催化从儿茶酚合成吡啶甲酸”。
- DOI:
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- 影响因子:0
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A.J.L.Cooper: "Ammonia metabolism in mammals:Interorgan relationships." Adv.Exp.Biol.Med.341. 21-37 (1993)
A.J.L.Cooper:“哺乳动物的氨代谢:器官间关系。”
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- 影响因子:0
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ASANO Yasuhisa其他文献
ASANO Yasuhisa的其他文献
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{{ truncateString('ASANO Yasuhisa', 18)}}的其他基金
Enzymatic synthesis of useful chemicals from nitrogen-containg substrates
从含氮底物酶法合成有用化学品
- 批准号:
23248015 - 财政年份:2011
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Use of the enzymes in the microbial and plant "aldoxime-nitrile pathway"for chiral synthesis
利用微生物和植物“醛肟-腈途径”中的酶进行手性合成
- 批准号:
20380053 - 财政年份:2009
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Discovery of amino acid amide racemase and its use in the dynamic kinetic resolution of amino acid amides
氨基酸酰胺消旋酶的发现及其在氨基酸酰胺动态拆分中的应用
- 批准号:
18380061 - 财政年份:2006
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Comparative Biochemistry of the Aldoxime-Nitrile Pathway of Plant and Microbial Origins
植物和微生物来源的醛肟-腈途径的比较生物化学
- 批准号:
16380064 - 财政年份:2004
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Discovery of Microbial "Aldoxime-Nitrile Pathway" and Its Application to Nitrile Synthesis
微生物“醛肟-腈途径”的发现及其在腈合成中的应用
- 批准号:
13460045 - 财政年份:2001
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Alternation in Substrate Specificity of Amino Acid Dehydrogenases based on the Structural Analyses
基于结构分析的氨基酸脱氢酶底物特异性的改变
- 批准号:
09460052 - 财政年份:1997
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Reaction Mechanisms and Application of New Bacterial Enzymes
新型细菌酶的反应机理及应用
- 批准号:
08044218 - 财政年份:1996
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for international Scientific Research
Development of Novel Lyases and Their Application to the Synthesis of Optically Active Compounds
新型裂解酶的开发及其在光学活性化合物合成中的应用
- 批准号:
07660117 - 财政年份:1995
- 资助金额:
$ 2.56万 - 项目类别:
Grant-in-Aid for Scientific Research (C)