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496 results on '"Sakuraba, Haruhiko"'

38. Gene cloning and characterization of the very large NAD-dependent L-glutamate dehydrogenase from the psychrophile Janthinobacterium lividum, isolated from cold soil

Author

Kawakami, Ryushi, Sakuraba, Haruhiko, and Ohshima, Toshihisa

Subjects
Cloning -- Research, Glutamate dehydrogenase -- Research, Biological sciences
Abstract

NAD-dependent L-glutamate dehydrogenase (NAD-GDH) activity was detected in cell extract from the psychrophile Janthinobacterium lividum UTB1302, which was isolated from cold soil and purified to homogeneity. The native enzyme (1,065 kDa, determined by gel filtration) is a homohexamer composed of 170-kDa subunits (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Consistent with these findings, gene cloning and sequencing enabled deduction of the amino acid sequence of the subunit, which proved to be comprised of 1,575 amino acids with a combined molecular mass of 169,360 Da. The enzyme from this psychrophile thus appears to belong to the GDH family characterized by very large subunits, like those expressed by Streptomyces clavuligerus and Pseudomonas aeruginosa (about 180 kDa). The entire amino acid sequence of the J. lividum enzyme showed about 40% identity with the sequences from S. clavuligerus and P. aeruginosa enzymes, but the central domains showed higher homology (about 65%). Within the central domain, the residues related to substrate and NAD binding were highly conserved, suggesting that this is the enzyme's catalytic domain. In the presence of NAD, but not in the presence of NADP, this GDH exclusively catalyzed the oxidative deamination of L-glutamate. The stereospecificity of the hydride transfer to NAD was pro-S, which is the same as that of the other known GDHs. Surprisingly, NAD-GDH activity was markedly enhanced by the addition of various amino acids, such as L-aspartate (1,735%) and L-arginine (936%), which strongly suggests that the N- and/or C-terminal domains play regulatory roles and are involved in the activation of the enzyme by these amino acids.

Published
2007

43. Novel archaeal alanine:glyoxylate aminotransferase from Thermococcus litoralis

Author

Sakuraba, Haruhiko, Kawakami, Ryushi, Takahashi, Hajime, and Ohshima, Toshihisa

Subjects
Alanine aminotransferase -- Research, Aminotransferases -- Research, Biological sciences
Abstract

A novel alanine:glyoxylate aminotransferase was found in a hyperthermophilic archaeon, Thermococcus litoralis. The amino acid sequence of the enzyme did not show a similarity to any alanine:glyoxylate aminotransferases reported so far. Homologues of the enzyme appear to be present in almost all hyperthermophilic archaea whose whole genomes have been sequenced.

Published
2004

44. Highly stable L-Lysine 6-Dehydrogenase from the thermophile Geobacillus stearothermophilus isolated from a Japanese hot spring: characterization gene cloning and sequencing, and expression

Author

Heydari, Mojgan, Ohshima, Toshihisa, Nunoura-Kominato, Naoki, and Sakuraba, Haruhiko

Subjects
Bacteria, Thermophilic -- Influence, Gene expression -- Research, Biological sciences
Abstract

L-Lysine dehydrogenases catalyze the oxidative deamination of L-Lysine in the presence of NAD and so far two types of dehydrogenases were discovered. Thermophile and hyperthermophilic microorganisms were screened for a more stable form of L-Lysine dehydrogenase and results are discussed.

Published
2004

47. Purification, characterization, and application of a novel dye-linked L-proline dehydrogenase from a hyperthermophilic archaeon, Thermococcus profundus

Author

Sakuraba, Haruhiko, Takamatsu, Yoshinori, Satomura, Takenori, Kawakami, Ryushi, and Ohshima, Toshihisa

Subjects
Biosensors -- Research, Dehydrogenases -- Research, Microbial enzymes -- Research, Bacteria, Thermophilic -- Research, Biological sciences
Abstract

Researchers describe a dye-linked dehydrogenase from a thermophilic bacterium that reduces 2,6-dichloroindophenol in the presence of L-proline. This enzyme could be useful in biosensors and may be more stable than its counterpart in mesophilic bacteria.

Published
2001

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