• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Studies on the Novel Enzyme Phenylalanine Dehydrogenase.

Research Project

Project/Area Number 05044138
Research Category

Grant-in-Aid for international Scientific Research

Allocation TypeSingle-year Grants
SectionJoint Research
Research InstitutionToyama Prefectural University

Principal Investigator

ASANO Yasuhisa  Associate Professor, Toyama Prefectural University, 工学部, 助教授 (00222589)

Co-Investigator(Kenkyū-buntansha) YAMADA Hideaki  Professor, Toyama Prefectural University, 工学部, 教授 (30027180)
KATO Yasuo  Assistant, Toyama Prefectural University, 工学部, 助手 (20254237)
クーパー アーサー.J.  米国コーネル大学, 医学部, 教授
COOPER Arthur j.l.  Professor, Cornell University Medical College
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1994: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1993: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsAmino Acid / Immunotitration / Amino Acid Dehydrogenase / Primary Structure / Enzyme Purification / Phenylalanine Dehydrogenase / Screening / Sequence Homology
Research Abstract

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.

Report

(2 results)
  • 1994 Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (43 results)

All Other

All Publications (43 results)

  • [Publications] M.Ueda: "The Substrate Specificity of Maleate Hydratase from Arthrobacter sp.strain MCI2612." Applied Microbiology and Biotechnology. 41. 215-218 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Asano: "3-Methylaspartase from a Facultative Anaerobe,Escherichia coli strain YG1002." FEMS Microbiology Letters. 118. 255-258 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Asano: "Catechol-2,3-Dioxygenase-catalyzed Synthesis of Picolinic Acids from Catechols." Bioscience Biotechnology and Biochemistry. 58. 2054-2056 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Asano: "Enzymes Involved in Theobromine Production from Caffeine by Pseudomonas putida No.352." Bioscience Biotechnology and Biochemistry. 58. 2303-2304 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kato: "Purification and Properties of Crystalline 3-Methylaspartate ammonia-lyase from Two Facultative Anaerobes,Citrobacter sp.Strain YG-0504 and Morganella morganii Strain YG-0601." Bioscience Biotechnology and Biochemistry. 59. 93-99 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kato: "3-Hydroxy-2-oxaspiro[4.5]-dec-3-ene,A New Inhibitor of Lactate Dehydrogenase." Tetrahedron Letters. (in press). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Sheu K-FR: "Abnormality of the α-ketoglutarate dehydrogenase complex in fibroblasts from familial Alzheimer's disease." Annals in Neurology. 35. 312-318 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ali G: "Isolation,characterization,and mapping of gene encoding dihydrolipoyl succinyltransferase(E2k) of human α-ketoglutarate dehydrogenase complex." Somatic Cell Molecular Genetics. 20. 99-105 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Abraham DG: "Characterization of a high molecular weight cysteine-S-conjugate β-lyase from rat kidney with activity toward leukotriene E4." Journal of Biological Chemistry. 270. 180-188 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Makar TK: "Lipid metabolism and membrane composition are altered in the brains of type II diabetic mice." Journal of Neurochemistry. (in press). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Makar TK: "Carnitine,carnitine acetyl translerase,and glutathione in Alzheimer brain." Neurochemical Research. (in press). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Makar TK: "Antioxidant defense systems in the brains of type II diabetic mice." Journal of Neurochemistry. (in press). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Cooper AJL: "Glutamine transaminase K-ω-amidase pathway in brain.In "Advances in Hepatic Encephalopathy and Nitrogen Exchange"" CRC Press,Boca Raton,Florida(in press), (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Cooper AJL: "Role of the liver in amino acid metabolism.In "Hepatology:A Textbook of Liver Disease 3rd Edition"" WB Saunders,Philadelphia(in press), (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Asano and Y.Kato: "Crystalline 3-Methylaspartase from a Facultative Anaerobe, Escherichia coli strain YG1002." FEMS Microbiology Letters. 118. 255-258 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Asano, T.Komeda and H.Yamada: "Enzymes Involved in Theobromine Production from Caffeine by Pseudomonas putida No.352." Bioscience Biotechnology and Biochemistry. 58. 2303-2304 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kato and Y.Asano: "Purification and Properties of Crystalline 3-Methylaspartate ammonia-lyase from Two Facultative Anaerobes, Citrobacter sp. Strain YG-0504 and Morganella morganii Strain YG-0601." Bioscience Biotechnology and Biochemistry. 59. 93-99 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Y.Kato, Y.Asano and A.J.L.Cooper: "3-Hydroxy-2-oxaspiro [4,5] -dec-3-cne, A New Inhibitor of Lactate Dehydrogenase." Tetrahedr on Letters. (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Sheu K-FR,Cooper AJL,Koike K,Koike M,Lindsay Jo, and Blass W.: "Abnormality of the alpha-ketoglutarate dehydrogenase complex in fibroblasts from familial Alzheimer's disease." Annals in Neurology. 35. 312-318 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Ali G,Wasco W,Cai X,Szabo P,Sheu K-FR,Cooper AJL,Oaston SM,Ousell JF,Tanzi RE,and Blass JP.: "Isolation, characterization, and mapping of gene encoding dihydrolipoyl succiny ltransferase (E2k) of human alpha-ketoglutarate dehydrogenase complex." Somatic Cell Molecular Genetics. 20. 99-105 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Makar TK,Hungund BL,Cook CA,Kashfi K,and Cooper AJL.: "Lipid metabolism and membrane composition are altered in the brains of type II diabetic mice." Journal of Neurochemisty. (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 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)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Makar TK,Rimpel-Lamhaouar K,Abraham DO,Ookhale VS,and Cooper AJL.: "Antioxidant defense systems in the brains of type II diabetic mice." Journal of Neurochemisty. (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Cooper AJL,and Abraham DO: CRC Press, Boca Raton, Florida (in press). Glutamine transaminase K-omega-amidase pathway in brain. in "Advances in Hepatic Encephalopathy and Nitrogen Exchange", (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Cooper AJL.: WB Saunders, Philadelphia(in press). Role of the liver in amino acid metabolism. in "Hepatology : A Textbook of Liver Disease 3rd Edition.", (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Ueda: "Purification and Chracterization of Maleate Hydratase from Arthrobacter sp.strain MCI2612" Bioscience Biotechnology Biochemistry. 57. 1545-1548 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Asano: "Occurrence of 3-Methylaspartate ammonia-lyase in Facultative Anaerobes and Their Application to Synthesis of 3-Substituted(S)-Aspartic Acids." Bioscience Biotechnology Biochemistry. 58. 223-224 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] M.Ueda: "Production of S-(+)-Citramalate from Itaconateby Resting Cells of Alcaligenes denitrificans strain MCI2775" Applied Microbiology and Biotechnology. 40. 466-469 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Asano: "Microbial Production of Theobromine from Caffeine" Bioscience Biotechnology Biochemistry. 57. 1286-1289 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Asano: "Microbial Production of D-Malate from Maleate" Applied and Environmental Microbiology. 59. 1110-1113 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Kato: "Enzymatic Synthesis of L-β-chloroalanine Using Amino Acid Dehydrogenase" Applied Microbiology and Biotechnology. 39. 301-304 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] A.J.L.Cooper: "Enzymology of cysteine S-conjugate beta-lyases" Advances in Pharmacology. 27. 73-115 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] A.J.L.Cooper: "High activities of glutamine transaminase K(dichloro vinylcysteine beta-lyase)and omega-amidase in the choroid plexus of rat brain" Journal of Neurochemistry. 61. 1731-1751 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] A.J.L.Cooper: "Ammonia metabolism in mammals:Interorgan relationships." Adv.Exp.Biol.Med.341. 21-37 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.K.Makar: "Vitamin E,glutathione,glutathione disulfide and enzymes of glutathione metabolism in cultures of chick astrocytes and neurons:Evidence that astrocytes play an important role in antioxidative processes in the brain." Journal of Neurochemistry. 62. 45-53 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] T.K.Makar: "Glutamine transaminase K and omega-amidase in primary cultures of astrocytes and neurons and in embryonic chick forebrain.Marked induction of brain glutamine transaminase K at time of hatching." Journal of Neurochemistry. 62. 1983-1988 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Y.Asano: "A New Enzyme D-Aminopeptidase:Structure,Function and Application to Organic Synthesis in Frontiers and New Horizons in Amino Acid Research" Elsevier, (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] A.J.L.Cooper: "Glutathione in the brain;disorders of glutathione metabolism.In:The Molecular and Genetic Basis of Neurological Disease" Butterworth-Heinemman,Boston,Massachusetts, (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Q.R.Smith: "Amino acid transport in brain.In:Mammalian Amino Acid Transport:Mechanism and Control" Plenum Press,New York, (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] W.A.Pulsinelli: "Metabolic encephalopathies and coma.,In:Basic Neurochemistry:Molecular,Cellular and Medical Aspects,5th Edition" Raven Press,New York, (1994)

    • Related Report
      1993 Annual Research Report

URL: 

Published: 1993-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi