Summary of Research Project Results Under the JSPS FY2000
"Research for the future Program"



1.Research Institution Osaka Medical College
 
2.Research Area Life Sciences
 
3.Research Field Structure and Functional Control Mechanism of Biomolecules
 
4.Term of Project FY1996〜FY2000
 
5.Project Number 96L00506
 
6.Title of Project Exploring the steric superfine structures and dynamics as the basis of substrate recognition and catalysis of multifunctional enzymes

7.Projetct Leader
Name Institution,Department Title of Position
Hiroyuki Kagamiyama Osaka Medical College, Biochemistry Professer

8.Core Members

Names Institution,Department Title of Position
Hideyuki Hayashi Osaka Medical College, Biochemistry Associate Professor

9.Cooperating Researchers

Names Institution,Department Title of Position
Ken Hirotsu Osaka City Univ. Graduate School of Science Professor
Retsu Miura Kumamoto Univ. School of Medicine Professor
Tetsuo Toraya Okayama Univ. Engineering Professor

10.Summary of Research Results

In this project, the three dimensional structures of pyridoxal enzymes (10), flavin enzymes (2), a cobalamin enzyme, and a quinoenzyme have been analyzed. Based on the structures, substrate-recognition and reaction mechanisms of these enzymes have been studied in detail, and important new findings were obtained. 1) A new concept for enzyme catalysis was presented. Enzymes are charged with strain energy, mainly found in the structures formed between the coenzymes and the enzyme proteins in their unliganded forms, and release the strain energy on binding of substrates, thereby reducing the activation energy barrier and enhancing the catalytic efficiency and stereoselectivity. The conformational changes of the enzymes responsible for the release of the strains have been elucidated structurally. 2) Quantum-chemical interpretation of the proton-transfer and electron-transfer process clarified how the enzyme protein controls the reaction specificity. In many enzymes, the reaction mechanism has been revised based on these findings of this project, which could not have been obtained by conventional methods of enzymology. 3) Several enzymes were found to undergo reorganization of the active site to accept different kinds of substrates, while maintaining the rigid structure of the part facing the reaction center. Based on this principle, substrate specificity could be altered by directed evolution using auxotrophy and DNA shuffling. The same strategy was successfully applied to develop a thermostable enzyme. 4) The process of formation of the prosthetic group (built-in coenzyme) has been elucidated at the atomic level. Other findings are as follows. 5) Discovery and gene cloning of a protein that catalyzes the energy-dependent replacement of the inactivated coenzyme in the enzyme with the intact adenosylcobalamin. 6) Isolation and gene cloning of enzymes from psychrophiles. X-ray structural analyses are under way to elucidate how these enzymes are active at low temperatures.

11.Key Words

(1)Pyridoxal enzymes 、(2)Flavin enzymes、(3)Cobalamin enzymes
(4)Build-in coenzymes、(5)Substrat recognition、(6)Enzyme-reaction mechanism
(7)X-ray crystallography、(8)Molecular design of active site、(9)Directed evolution

12.References

[Reference Articles]
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Yoshimura, T. Stereospecificity for the hydrogen transfer and molecular evolution of pyridoxal enzyme.
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J. Biol. Chem. 271 1996 9612-9618

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Yamamoto, N. Cloning sequencing expression of the UvrA gene from an extremely thermophilic bacterium, Thermus thermophilus HB8.
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Mizutani, H. Three-dimensional structure of porcine kidney D-amino acid oxidase at 3.0 Å.
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J. Biochem. 120 1996 14-17

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Setoyama, C. Crystallization of expressed porcine kidnet D-amino acid oxidase and preliminary X-ray crystallographic characterzation.
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J. Biochem. 119 1996 1114-1117

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Nakamura, N. Biosynthesis of Topa quinone cofactor in bacterial amino oxidases. Solvent origin of C-2 oxygen determined by Raman spectroscopy.
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J. Biol. Chem. 271 1996 22598-22603

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Tobimatsu, T. Cloning, sequencing and high-level expression of the genes encoding adenosylcobalamin-dependent glycerol dehydrase of Klebsiella pneumoniae.
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Biochemistry 35 1996 6754-6761

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Biochim. biophys. Acta. 1308 1996 189-192

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Mollova, E. T. Use of 1H-15Nheteronuclear multiple-quantum coherence NMR spectroscopy to study the active site of aspartate aminotransferase.
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Biosci. Biotech. Biochem. 61 1997 410-417

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Biochemistry 36 1997 1953-1959

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Toraya, T. An electron paramagnetic resonance study on the mechanism-based inactivation of adenosylcobalamin-dependent diol dehydrase by glycerol and other substrates.
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Biochim. Biophys. Acta. 1337 1997 11-16

Author Title of Article
Oue, S. Paracoccus denitrificans aromatic amino acid aminotransferase: A model enzyme for the study of dual substrate recognition mechanism.
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J. Biochem. 121 1997 161-171

Author Title of Article
Galkin, A. Conversion of α-keto acid to D-amino acid by coupling of four enzyme reactions.
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J. Ferment. Bioeng. 83 1997 299-300

Author Title of Article
Hayashi, H. Transient - state kinetics of the reaction of aspartate aminotransferase with aspartate at low pH reveals dual routes in the enzyme - substrate association process.
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Biocnemistry 36 1997 13558-13569

Author Title of Article
Hazekawa, I A Raman study on the C(4)=0 stretching mode of flavins in flavoenzymes: Hydrogen bonding at the C(4)=0 moiety.
Journal Volume Year Pages Concerned
J. Biochem. 121 1997 1147-1154

Author Title of Article
Kawaguchi, S. Enzyme flexibility: New concept in recognition of hydrophobic substrates.
Journal Volume Year Pages Concerned
J. Biochem. 122 1997 55-63

Author Title of Article
Kishimoto, K. Mutation of Arg98, which serves as a substrate - recognition site of D-amino acid aminotransferase, can be partly compensated for by mutation of Tyr88 to an argrinine residue
Journal Volume Year Pages Concerned
J. Biochem. 122 1997 1182-1189

Author Title of Article
Mihara, H. Cysteine sulfinate desulfinase, a NIFS like protein of Escherichia coli with selenocysteine lyase and cysteine desulfurase activities
Journal Volume Year Pages Concerned
J. Biol. Chem. 272 1997 22417-22424

Author Title of Article
Miura, R. Structural and mechanistic studies on D-amino acid oxidase - substrate complex: Implications of the crystal structure of enzyme - substrate analog complex.
Journal Volume Year Pages Concerned
J. Biochem. 122 1997 825-833

Author Title of Article
Mori, K. A protein factor is essential for in situ reactivation of glycerol- inactivated adenosylcobalamin-dependent diol dehydrase.
Journal Volume Year Pages Concerned
Biosci. Biotech. Biochem. 61 1997 1729-1733

Author Title of Article
Mori, K. Characterization, sequencing and expression of the genes encoding a reactivating factor for glycerol - inactivated adenosylcobalamin - degendent diol dehydratase.
Journal Volume Year Pages Concerned
J. Biol. Chem. 272 1997 32034-32041

Author Title of Article
Mure, M. Chemical and biochemical characteristics of topaquinone.
Journal Volume Year Pages Concerned
Biosci. Biotech. Biochem. 61 1997 410-417

Author Title of Article
Nakamura, N. Topaquinone - dependent amine oxidase. Identification of reaction intermediates by Raman spectroscopy.
Journal Volume Year Pages Concerned
Biochemistry 36 1997 11479-11486

Author Title of Article
Nishino, J. An anomalous side reaction of the Lys303 mutant aromatic - L - amino acid decarboxylase unravels the role of the residue in catalysis.
Journal Volume Year Pages Concerned
J. Biochem. 121 1997 604-611

Author Title of Article
Oikawa, T. A novel of D - mannitol dehydrogenase from Acetobacter xylinum : Occurrence, purification, and basic properties
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Biosci. Biotech. Biochem. 61 1997 1778-1782

Author Title of Article
Okada, K. Three - dimensional structure of Escherichia coli branched - chain amino acid aminotransferase at 2.5 Å resolution.
Journal Volume Year Pages Concerned
J. Biochem. 121 1997 637-641

Author Title of Article
Ruggiero, C. E. Mechanistic studis of topa quinone biogenesis in phenylethylamine oxidase.
Journal Volume Year Pages Concerned
Biochemistry 36 1997 1953-1958

Author Title of Article
Setoyama, C. Structural and functional characterization of the fuman brain D - aspartate oxidase.
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J. Biochem. 121 1997 798-803

Author Title of Article
Shimizu, E. Purification and properties of phenylethylamine oxidase of Arthrobacter globiformis.
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Biosci. Biotech. Biochem. 61 1997 501-505

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Tamaoki, H. Spectroscopic studies of rat liver acyl - CoA oxidase with reference to recognition and activation of substrate.
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J. Biochem. 121 1997 1139-1146

Author Title of Article
Tobimatsu, T. Heterologous expression, purification and properties of diol dehydratase, an adenosylcobalanin - dependent enzyme of Klebsiella oxytoca.
Journal Volume Year Pages Concerned
Arch. Biochem. Biophys. 347 1997 132-140

Author Title of Article
Tanizawa, K. Molecular mechanism of topa quinone biogenesis in copper amine oxidase
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Kagaku to Seibutsu 35 1997 569-575

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Toraya, T. Current status of studies on vitamin B12 enzyme.
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Seikagaku 69 1997 1281-1287

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Wilce, M. C. Crystal structures of the copper - containing amine oxidase from Arthrobacter globiformis in the holo - and apo - forms: Implications for the biogenesis of topa quinone.
Journal Volume Year Pages Concerned
Biochemistry 36 1997 16116-16133

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Yamada, K. Purification and some properties of cobalamin - dependent methionine synthase from rat liver.
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J. Nutr. Sci. Vitaminol. 43 1997 177-186

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Ikushiro, H. Analysis of the pH - and ligand - induced spectral transitions of tryptophanase : Activation of the coenzyme at the early steps of the catalytic cycle.
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Biochemistry 37 1998 3043-3052

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Lim, Y. H. Non-stereospecific transamination catalyzed by pyridoxal - dependent amino acid racemases of broad substrate specificity.
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J. Biol. Chem. 273 1998 4001-4005

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Liu, L. Compensation for D-glutamate auxotrophy of IWM335 by D-amino acid aminotransferase gene and regulation of murl expression.
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Biosci. Biotech. Biochem. 62 1998 193-195

Author Title of Article
Gutierrez, A. A mutant D-amino acid aminotransferase with broad specificity: Construction by replacement of the interdomain loop Pro119 - Arg120 - Pro121 by Gly - Gly -Gly.
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Protein Engineering. 11(1) 1998 53-58

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Yamada, K. Cloning, Sequencing, and Heterologous Expression of Rat Methionine Synthase cDNA.
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Biosci. Biotech. Biochem. 61 1998 2155-2160

Author Title of Article
Yamanishi, M. EPR Spectroscopic Evidence for the Mechanism-based Inactivation of Adenosylcobalamin-dependent Diol Dehydratase by Coenzyme Analogs.
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J. Biochem. 124 1998 598-601

Author Title of Article
Tobimatsu, T. Molecular Cloning, Sequencing and Characterization of the Genes for Adenosylcobalamin-dependent Diol Dehydratase of Klebsiella pneumoniae.
Journal Volume Year Pages Concerned
Biosci. Biotech. Biochem. 62 1998 1774-1777

Author Title of Article
Yamanishi, M. Evidence for Axial Coordination of 5,6-Dimethylbenzimidazole to the Cobalt Atom of Adenosylcobalamin Bound to Diol Dehydratase.
Journal Volume Year Pages Concerned
Biochemistry 37 1998 4799-4803

Author Title of Article
Kawaguchi, S. Thermodynamics and Molecular Simulation Analysis of Hydrophobic Substrate Recognition by Aminotransferases.
Journal Volume Year Pages Concerned
J. Biol. Chem. 273 1998 18353-18364

Author Title of Article
Nakai, T. Crystallization and Preliminary X-ray Characterization of Aspartate Aminotransferase from an Extreme Thermophile, Thermus thermophilus HB.
Journal Volume Year Pages Concerned
Acta Crystallogr. D54(5) 1998 1032-1034

Author Title of Article
Okamoto, A. Crystal Structures of Paracoccus denitrificans Aromatic Amino Acid Aminotransferase: A Substrate Recognition Site Constructed by Rearrangement of Hydrogen Bond Network.
Journal Volume Year Pages Concerned
J. Mol. Biol. 280 1998 443-461

Author Title of Article
Nobe, Y. The Novel Substrate Recognition Mechanism Utilized by Aspartate Aminoransferase of the Extreme Thermophile Thermus thermophilus HB8.
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J. Biol. Chem. 273 1998 29554-29564

Author Title of Article
Oikawa, T. Endo- β -Glucanase Secreted by a Psychrotrophic Yeast: Purification and Characterization.
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Biosci. Biotech. Biochem. 62 1998 1751-1756

Author Title of Article
Yano, T. Directed Evolution of An Aspartate Aminotransferase With New Substrate Specificities.
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Proc. Natl. Acad. Sci. USA 95 1998 5511-5515

Author Title of Article
Ishii, S. Aromatic L-Amino Acid Decarboxylase: Conformational Change in The Flexible Region Around Arg334 Is Reguired During The Transaldimination Process.
Journal Volume Year Pages Concerned
Protein Sci. 7 1998 1802-1810

Author Title of Article
Hayashi, H. The Imine-Pyridine Torsion of The Pyridoxal 5'-Phosphate Schiff Base of Aspartate Aminotransferase Lowers Its pKa in The Unliganded Enzyme And Is Crucial For The Successive in The pKa During Catalysis.
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Biochemistry 37 1998 15076-15085

Author Title of Article
Li, Y.-F. Crystal Structures of Reaction Intermediates of L-2-Haloacid Dehalogenase And Implications for The Reaction Mechanism.
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J. Biol. Chem. 273 1998 15035-15044

Author Title of Article
Sugio, S. Crystal Structures of L201A Mutant of D-Amino Acid Aminotransferase at 2.0 A Resolution: Implication of Structural Role of Leu201 in Transamination.
Journal Volume Year Pages Concerned
Protein Engineering 11 1998 613-619

Author Title of Article
Uo, T. Occurrence Of Pyridoxal 5'-Phosphate-Dependent Serine Racemase in Silkworm, Bombyx mori.
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Biochem. Biophys. Res. Commun. 246 1998 31-34

Author Title of Article
Fuchikami, Y. Construction and Properties of a Fragmentary D-Amino Acid Aminotransferase.
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J. Biochem. 124 1998 905-910

Author Title of Article
Kurokawa, Y. Transamination as a Side-Reaction Catalyzed by Alanine Racemase of Bacillus stearothemophilus.
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J. Biochem. 124 1998 1163-1169

Author Title of Article
Li, Y. -F X-Ray Structure of A Reaction Intermediate of L-2-Haloacid Dehalogenase with L-2-Chloropropionamide.
Journal Volume Year Pages Concerned
J. Biochem. 124 1998 20-22

Author Title of Article
Nishina, Y. Resonance Raman Study on Reduced Flavin in Purple Intermediate of Flavoenzyme: Use of [4-Carbonyl-18O]-Enriched Flavin.
Journal Volume Year Pages Concerned
J. Biochem. 124 1998 200-208

Author Title of Article
Matsuzaki, R. Exploring A Channel to The Active Site of Copper / Topaquinone-Containing Phenylethylamine Oxidase by Chemical.
Journal Volume Year Pages Concerned
Biochemistry 37 1998 13947-13957

Author Title of Article
Oue, S. Redesignning The Substrate Specificity of An Enzyme by Cumulative Effects of The Mutations of Non-Active Site Residues.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 2344-2349

Author Title of Article
Toraya, T. A Reactivating Factor for Coenzyme B12-Dependent Diol Dehydratase.
Journal Volume Year Pages Concerned
J. Biol. Chem. 247 1999 3372-3377

Author Title of Article
Watanabe, A. Role of Lysine39 of Alanine Racemase from Bacillus Stearothermophilus That Binds Pyridoxal 5'-Phosphate:Chemical Rescue Studies of Lys39→Ala Mutant.
Journal Volume Year Pages Concerned
J. Biol. Chem. 247 1999 4189-4194

Author Title of Article
Tamaoki, H. Mechanism for Recognition and Activation of Substrate in Medium-Chain Acyl-CoA Dehydrogenase.
Journal Volume Year Pages Concerned
J. Biochem. 125 1999 285-296

Author Title of Article
Odagaki, Y. Crystal Structure of The Pyroglutamyl Peptidase I from Bacillus amyloliquefaciens Reveals New Structure for A Cysteine Protease
Journal Volume Year Pages Concerned
Structure 7 1999 399-411

Author Title of Article
Galkin, A. Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning. Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts.
Journal Volume Year Pages Concerned
Appl. Environ. Microbiol. 65(9) 1999 4014-4020

Author Title of Article
Mihara, H. AnifS-like Gene, csdB, Encodes an Escherichia coli Counterpart of Mammalian Selenocysteine Lyase.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274(21) 1999 14768-14772

Author Title of Article
Watanabe, A. Tyrosine 265 of Alanine Racemase Serves as a Base Abstracting a-Hydrogen from L-Alanine: The Counterpart Residue to Lysine 39 Specific to D-Alanine.
Journal Volume Year Pages Concerned
J. Biochem. 126(4) 1999 781-786

Author Title of Article
Tamaoki, H. Mechanism for recognition and activation of substrate in medium-chain acyl-CoA dehydrogenase.
Journal Volume Year Pages Concerned
J. Biochem. 125 1999 285-296

Author Title of Article
Hirota, S. Spectroscopic Characterzation of Carbon Monoxide Complexes Generated for Copper/Topa Quinone-Containing Amine Oxidases.
Journal Volume Year Pages Concerned
Biochemistry 38 1999 14256-14263

Author Title of Article
Tanizawa, K. Built-in coenzymes:amino acid residue-derived new coenzyme.
Journal Volume Year Pages Concerned
Tanpakushittsu ・ Kakusan ・ Kohso 44 1999 1947-1958

Author Title of Article
Yamada, K. Heterologous High-level Expression, Purification, and Enzymological Properties of Recombinant Rat Cobalamin-dependent Methionine Synthase.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 35571-35576

Author Title of Article
Mori, K. The Mechanism of Reactivation of Coenzyme B12-Dependent Diol Dehydratase by a Molecular Chaperone-Like Reactivating Factor.
Journal Volume Year Pages Concerned
Biochemistry 38 1999 13170-13178

Author Title of Article
Toraya, T. Direct Participaton of Potassium Ion in the Catalysis of Coenzyme B12-Dependent Diol Dehydratase.
Journal Volume Year Pages Concerned
J. Biochem. 126 1999 650-654

Author Title of Article
Masuda, J. Crystallization and Preliminary X-ray Study of Two Crystal Forms of Klebsiella oxytoca Diol Dehydratase-cyanocobalamin Complex.
Journal Volume Year Pages Concerned
Acta. Crystallogr. D55 1999 907-909

Author Title of Article
Shibata, N. A New Mode of B12 Binding and the Direct Particpation of a Potassium Ion in Enzyme Catalysis. X-ray Structure of Diol Dehydratase.
Journal Volume Year Pages Concerned
Structure 7 1999 997-1008

Author Title of Article
Tobimatsu, T. Identification and Expression of the Genes Encoding a Reactivating Factor for Adenosylcobalamin-dependent Glycerol Dehydratase.
Journal Volume Year Pages Concerned
J. Bacter. 181 1999 4410-4113

Author Title of Article
Oikawa, T. Production of D-Glutamate from L-Glutamate with Glutamate Racemase and L-Glutamate Oxidase.
Journal Volume Year Pages Concerned
Biosci. Biotech. Biochem. 63 1999 2168-2173

Author Title of Article
Oikawa, T. Stereoisomers of Glutathione: Preparation and Enzymatic Reactivities.
Journal Volume Year Pages Concerned
J. Nutr. Sci. Vitaminol. 45 1999 223-229

Author Title of Article
Nakai, T. Structure of Thermus thermophilus HB8 Aspartate Aminotransferase and Its Complex with Maleate.
Journal Volume Year Pages Concerned
Biochemistry 38(8) 1999 2413-2424

Author Title of Article
Hoseki, J. Directed Evolution of Thermostable Kanamycin-Resistant Gene Products in an Extremely Thermophilic Bacterium, Thermus thermophilus.
Journal Volume Year Pages Concerned
J. Biochem. 126(5) 1999 951-956

Author Title of Article
Hayashi, H. Acid-Base Chemistry of the Reaction. of Aromatic L-Amino Acid Decarboxylase and Dopa Analyzed by Transient and Steady-State Kinetics: Preferential Binding of the Substrate with Its Amino Group Unprotonated.
Journal Volume Year Pages Concerned
Biochemistry 38 1999 15615-15622

Author Title of Article
Oue, S. Cocrystallization of a Mutant Aspartate Aminotransferase with a C5-Dicarboxylic Substrate Analog: Structural Comparison with the Enzyme-C4-Dicarboxylic Analog Complex.
Journal Volume Year Pages Concerned
J. Biochem. 127 2000 337-343

Author Title of Article
Koyanagi, T. Molecular Cloning snd Heterologous Expression of Pea Seedling Copper Amine Oxidase.
Journal Volume Year Pages Concerned
Biosci. Biotech. Biochem. 64 2000 717-722

Author Title of Article
Mihara, H. Kinetic and mutational studies of three Nifs homologs from Escherichia coli : Mechanistic difference between L-cysteine desulfurase and L-selenocysteine lyase reactions.
Journal Volume Year Pages Concerned
J. Biochem. 127(2) 2000 559-567

Author Title of Article
Mihara, H. cDNA cloning, purification, and characterization of mouse liver selenocysteine lyase. Condidate for selenium delivery protein in selenoprotein synthesis.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275(9) 2000 6195-6200

Author Title of Article
Fujii, T. Structure of Nifs homologue: X-ray structure analysis of Csd B, an Escherichia coli counterpart of mammalian selenocysteine lyase.
Journal Volume Year Pages Concerned
Biochemistry 39(6) 2000 1263-1273

Author Title of Article
Gutierrez, A. Modulation of activity and substrate specificity by modifying the backbone length of the distant interdomain loop of D-amino acid aminotransferase.
Journal Volume Year Pages Concerned
Eur. J. Biochem. 267(24) 2000 7218-7223

Author Title of Article
Nakai, T. Three dimensional structure of 4-amino-4-deoxychorismate lyase from Escherichia coli.
Journal Volume Year Pages Concerned
J. Biochem. 128 2000 29-38

Author Title of Article
Jhee, K. -H. Stereochemistry of the transamination reaction catalyzed by aminodeoxychorismate lyase from Escherichia coli: Close relationship between fold type and stereochemistry.
Journal Volume Year Pages Concerned
J. Biochem. 128(4) 2000 679-686

Author Title of Article
Ishijima, J. Free energy requirement for domain movement of enzyme.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275(25) 2000 18939-18945

Author Title of Article
Matsui, I. The molecular structure of hyperthermostable aromatic aminotransferase with novel substrate specificity from Pyrococcus horikoshii.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275(7) 2000 4871-4879

Author Title of Article
Mizutani, H. Three-dimensional structure of the purple intermediate of porcine kidney D-amino acid oxidase. Optimization of the oxidative half-reaction through alignment of the product with reduced flavin.
Journal Volume Year Pages Concerned
J. Biochem. 128 2000 73-81

Author Title of Article
Toraya, T. Radical atalysis of B12 enzymes: structure, mechanism, inactivation and reactivation of diol and glycerol dehydratasesm.
Journal Volume Year Pages Concerned
Cell. Mol. Life Sci. 57 2000 106-127

Author Title of Article
Masuda, J. How a grotein generates a catalytic radical from coenzyme B12 : X-ray structure of a diol - dehydratase - adeninylpentylcobalamin Complex.
Journal Volume Year Pages Concerned
Structure 8 2000 775-788

Author Title of Article
Tobimatsu, T. Specificities of reactivating factors for adenosylcoblamin - dependent diol dehydratase and glycerol dehydratase.
Journal Volume Year Pages Concerned
Arch. Microbiol. 174 2000 81-88

Author Title of Article
Nishina, Y. Substrate recognition and activation mechanism of D-amino acid oxidase : A study using substrate analogs.
Journal Volume Year Pages Concerned
J. Biochem. 128 2000 213-223

Author Title of Article
M. M. Islam The substrate activation process in the catalytic reaction of Escherichia coli aromatic amino acid aminotransferase.
Journal Volume Year Pages Concerned
Biochemistry 39(50) 2000 15418-15428

Author Title of Article
Kagamiyama, H. Studies of transaminases on the basis of their tertiary structures
Journal Volume Year Pages Concerned
Vitamins 74(1) 2000 1-12

Author Title of Article
Ura, H. Temperature dependence of the enzyme - substrate recognition mechanism.
Journal Volume Year Pages Concerned
J. Biochem. 129(1) 2001 173-178

Author Title of Article
Mizuguchi, H. Strain is more important than electrostatic interaction in controlling the pKa of the catalytic group in aspatate aminotransferase.
Journal Volume Year Pages Concerned
Biochemistry 40 2001 353-360

Author Title of Article
Miura, R. Porcine kidney D-amino acid oxidase : The three dimensional structure and its catalytic mechanism based on the enzyme - substrate complex model.
Journal Volume Year Pages Concerned
J. Mol. Catalysis. B, Enzymatic 12 2001 43-52

Author Title of Article
Haruyama, K. Structure of Escherichia coli histidinol - phosphate aminotransferase and its complexes with histidinol - phosphate and N-(5'-phosphopyridoxyl)-L-glutamate double substrate recognition of the enzyme
Journal Volume Year Pages Concerned
Biochemistry 40 2001 in press

Author Title of Article
Miura, R. Versatility and specificity of flavoenzymes : Control mechanism of flavin reactivity.
Journal Volume Year Pages Concerned
The Chemical Record 1 2001 in press

Author Title of Article
Watanabe, A. Stereochemistry of the hydrogen abstraction from pyridoxamine phosphate catalyzed by alanine racemase of Bacillus stearothermophilus.
Journal Volume Year Pages Concerned
J. Mol. Catalysis   2001 in press

Author Title of Article
Eda, M. Theoretical Predictions on the Mechanism of Catalysis of Coenzyme B12 - Dependent Diol.
Journal Volume Year Pages Concerned
Dehydratase   2001 in press

Author Title of Article
Oikawa, T. Fragmentary form of thermostable leucine dehydrogenase of Bacillus stearothermophilus: Its construction and reconstruction of active fragmentary enzyme.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun   2001 in press

Author Title of Article
Oikawa, T. Alanine aminotransferase II from the hyperthermophilic archaeon Thermococcus sp. TK1: Purification, molecular characterization, and expression in Escherichia coli.
Journal Volume Year Pages Concerned
Eur, J. Biochem.   2001 in press

Author Title of Article
Oikawa, T. Psychrophilic valine dehydrogenase of Antarctic Psychophile, Cytophaga sp. P1: Purification, molecular characterization, and expression in Escherichia coli.
Journal Volume Year Pages Concerned
Eur. J. Biochem.   2001  

Author Title of Article
Kazuoka, T. Thermostable alcohol dehydrogenase from a Psycophile, Cytophage sp. KUC-1: Cloning, molecular characterization.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA   2001 in press

Author Title of Article
Oikawa, T Alanine aminotransferase I from the hyperthermophilic Archaeon Thermococcus sp. TK1: Purification, molecular characterization, and expression in Escherichia coli.
Journal Volume Year Pages Concerned
Eur. J. Biochem.   2001 in press

[Reference Books]
Author Title of Book
Miura, R. Flavins and Flavoproteins
Publisher Year Pages
Agency for Scientific Publ., Berlin. 1999 939

Author Title of Book
Setoyama, C. Flavins and Flavoproteins
Publisher Year Pages
Agency for Scientific Publ., Berlin. 1999 939

Author Title of Book
Kagamiyama, H. Methodes in Enzymology (Branched. chain amino acid aminotransferase in E. coli)
Publisher Year Pages
Academic Press 2000 550

Author Title of Book
Hayashi, H. Biochemistry and Molecular Bioligy of Vitamin B6 and PQQ - dependent proteins
Publisher Year Pages
Birkhauser Verlag 2000 376

Author Title of Book
Hirotsu, K. Biochemistry and Molecular Biology of Vitamin B6 and PQQ - dependent proteins.
Publisher Year Pages
Birkhauser Verlag 2000 376


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