A5087777648- Enzyme Structure and Function
- Amino Acid Enzymes and Metabolism
- Biochemical and Molecular Research
- Metabolism and Genetic Disorders
- Enzyme Catalysis and Immobilization
- Polyamine Metabolism and Applications
- Diet, Metabolism, and Disease
- Genomics and Phylogenetic Studies
- Protein Structure and Dynamics
- Electrochemical sensors and biosensors
- Microbial Metabolic Engineering and Bioproduction
- GABA and Rice Research
- Crystallization and Solubility Studies
- Enzyme Production and Characterization
- Mass Spectrometry Techniques and Applications
- X-ray Diffraction in Crystallography
- Chemical Synthesis and Analysis
- Biopolymer Synthesis and Applications
- Pancreatic function and diabetes
- Probiotics and Fermented Foods
- Metabolomics and Mass Spectrometry Studies
- Bacterial Genetics and Biotechnology
- Electrochemical Analysis and Applications
- Biofuel production and bioconversion
- Cancer, Hypoxia, and Metabolism
Osaka Institute of Technology
2013-2022
Kyushu University
2007-2018
Genetic Resources Center
2009-2013
Tokushima University
1999-2006
Kyoto University of Education
1982-2005
Osaka University
2005
Kansai Medical University
2005
Kirin (Japan)
2002
The University of Tokyo
2002
Tokyo Institute of Technology
2002
The distribution of bacterial leucine dehydrogenase (L-leucine:NAD+ oxidoreductase, deaminating, EC 1.4.1.9) was investigated, and Bacillus sphaericus (IFO 3525) found to have the highest activity enzyme. Leucine dehydrogenase, which purified homogeneity crystallized from B. sphaericus, has a molecular weight about 245,000 consists six identical subunits (Mr = 41,000). enzyme catalyzes oxidative deamination L-leucine, L-valine, L-isoleucine, L-norvaline, L-alpha-aminobutyrate, L-norleucine,...
Levels of free D-amino acids were compared in 11 vinegars produced from different sources or through manufacturing processes. To analyze the D- and L-amino acids, enantiomers initially converted into diastereomers using pre-column derivatization with o-phthaldialdehyde plus N-acethyl-L-cysteine N-tert-butyloxycarbonyl-L-cysteine. This was followed by separation resultant fluorescent isoindol derivatives on an octadecylsilyl stationary phase ultra-performance liquid chromatography. The...
Genes encoding 2-deoxy-d-ribose-5-phosphate aldolase (DERA) homologues from two hyperthermophiles, the archaeon Pyrobaculum aerophilum and bacterium Thermotoga maritima, were expressed individually in Escherichia coli, after which structures activities of enzymes produced characterized compared with those E. coli DERA. To our surprise, hyperthermophilic DERAs showed much greater catalysis sequential aldol condensation using three acetaldehydes as substrates than enzyme, even at a low...
Thermophiles have important advantages over mesophiles as host organisms for high-temperature bioprocesses, functional production of thermostable enzymes, and efficient expression enzymatic activities in vivo. To capitalize on these thermophiles, we describe here a new inducible gene system the thermophile Geobacillus kaustophilus HTA426. Six promoter regions HTA426 genome were identified analyzed profiles using β-galactosidase reporter assay. This analysis region upstream putative...
A filamentous bacteriophage, φOH3, was isolated from hot spring sediment in Obama Japan with the hyperthermophilic bacterium Thermus thermophilus HB8 as its host. Phage which classified into Inoviridae family, consists of a flexible particle 830 nm long and 8 wide. φOH3 stable at temperatures ranging 70 to 90°C pHs 6 9. one-step growth curve phage showed 60-min latent period beginning immediately postinfection, followed by intracellular virus production during subsequent 40 min. The released...
A gene encoding an ADP-dependent phosphofructokinase homologue has been identified in the hyperthermophilic archaeon Methanococcus jannaschii via genome sequencing. The encoded a protein of 462 amino acids with molecular weight 53,361. deduced acid sequence showed 52 and 29% identities to glucokinase from Pyrococcus furiosus, respectively. was overexpressed Escherichia coli, produced enzyme purified characterized. To our surprise, high activities for both phosphofructokinase. native mass...
The gene encoding the thermostable phenylalanine dehydrogenase [EC 1.4.1.-] of a thermophile, Thermoactinomyces intermedius, was cloned and its complete DNA sequence determined. (pdh) consists 1,098 nucleotides encodes 366 amino acid residues corresponding to subunit (Mr 41,000) hexameric enzyme. deduced from nucleotide pdh T. intermedius 56.0 42.1% homologous those dehydrogenases Bacillus sphaericus Sporosarcina ureae, respectively. It shows 47.5% homology that leucine B....
Leucine dehydrogenase has been purified to homogeneity from a moderate thermophilic actinomycete, Thermoactinomyces intermedius IFO 14230. The enzyme can be stored without loss of its activity at low temperature (e.g., 4°C) for over two years. was more thermostable higher concentrations salts such as NaCl and KCl. It retained about 90% on incubation 70°C least 40 min in the presence 3 M NaCl. Michaelis constants NAD, l ‐leucine, NADH, 2‐oxoisocaproate ammonia were determined 0.36, 2.0,...
We screened various thermophiles for meso-diaminopimelate dehydrogenase (meso-DAPDH, EC 1.4.1.16), which catalyzes the NAD(P)-dependent oxidative deamination of meso-diaminopimelate, and found enzyme in a thermophilic bacterium isolated from compost Japan. The grew well aerobically at around 55°C was identified as Ureibacillus thermosphaericus strain A1. purified about 47-fold to homogeneity crude cell extract using five successive purification steps. molecular mass protein 80 kDa, molecule...
Phenylalanine dehydrogenase (L-phenylalanine:NAD oxidoreductase, deaminating; EC 1.4.1.-) was found in various thermophilic actinomycetes. We purified the enzyme to homogeneity from Thermoactinomyces intermedius IFO 14230 by heat treatment and Red Sepharose 4B, DEAE-Toyopearl, CL-4B, Sephadex G-100 chromatographies with a 13% yield. The relative molecular weight of native estimated be about 270,000 gel filtration. consists six subunits identical (41,000) is highly thermostable: it not...
Although ATP is the most common phosphoryl group donor for kinases, some kinases from certain hyperthermophilic archaea such as Pyrococcus horikoshii and Thermococcus litoralis use ADP donor. Those are ADP-dependent glucokinases (ADPGK) phosphofructokinases in their glycolytic pathway. Here, we succeeded gene cloning ADPGK P. OT3 (phGK) Escherichia coli,and easy preparation of enzyme, crystallization, structure determination apo enzyme. Recently, three-dimensional T. (tlGK) a complex with...
Two novel types of dye-linked l-proline dehydrogenase complex (PDH1 and PDH2) were found in a hyperthermophilic archaeon, Pyrococcus horikoshii OT3. Here we report the first crystal structure PDH1, which is heterooctameric (αβ)4 containing three different cofactors: FAD, FMN, ATP. The was determined by x-ray crystallography to resolution 2.86 Å. β subunit, an catalytic component FAD as cofactor, similar that monomeric sarcosine oxidase. On other hand, α subunit possessed unique composed...
Abstract Alanine dehydrogenase catalyzed the conversion of 3‐fluoropyruvate into 3‐fluoro‐ L ‐alanine in presence NADH and ammonia. The optimum pH reaction was 7.8. K m values enzyme for 3‐fluoropyruvate, polyethylene glycol‐bound NADH, ammonia were 2.94, 0.56, 105m M , respectively. 3‐Fluoro‐ selectively continuously produced from ammonium formate an membrane reactor by multienzyme system alanine with a simultaneous coenzyme regeneration. average space–time yield 73% 75 g/L day,...