徳島大学 教育・研究者情報データベース(EDB)

1. 徳島大学.工学部.生物工学科.生物反応工学講座(〜2023年3月31日)
2. 独立行政法人 産業技術総合研究所

1. (英) Shimizu Yasuhiro

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2. 櫻庭 春彦

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3. 川上 竜巳([徳島大学.大学院社会産業理工学研究部.生物資源産業学域.食料科学系.食料科学分野]/[徳島大学.生物資源産業学部])

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4. 郷田 秀一郎([長崎大学])

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5. 河原林 裕(独立行政法人産業技術総合研究所生物機能工学研究部門)

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6. 大島 敏久([九州大学])

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(英) L-Threonine dehydrogenase from the hyperthermophilic archaeon Pyrococcus horikoshii OT3

(英) gene cloning and enzymatic characterization

(英) A gene encoding the L-threonine dehydrogenase homologue has been identified in a hyperthermophlic archaeon Pyrococcus horikoshii OT3 via genome sequencing. The gene was cloned and expressed in Escherichia coli. The purified enzyme from the recombinant E. coli was extremely thermostable; the activity was not lost after incubation at 100 degrees C for 20 min. The enzyme (molecular mass: 192 kDa) is composed of a tetrameric structure with a type of subunit (41 kDa). The enzyme is specific for NAD and utilizes L-threonine, L-serine and DL-threo-3-phenylserine as the substrate. The enzyme required divalent cations such as Zn(2+), Mn(2+) and Co(2+) for the activity, and contained one zinc ion/subunit. The K(m) values for L-threonine and NAD at 50 degrees C were 0.20 mM and 0.024 mM, respectively. Kinetic analyses indicated that the L-threonine oxidation reaction proceeds via a random mechanism with regard to the binding of L-threonine and NAD. The enzyme showed pro-R stereospecificity for hydrogen transfer at the C4 position of the nicotinamide moiety of NADH. This is the first description of the characteristics of an L-threonine dehydrogenase from the archaea domain.

1. (英) Alcohol Oxidoreductases
2. (英) Amino Acid Sequence
3. (英) Animals
4. (英) Catalysis
5. (英) Cloning, Molecular
6. (英) Hydrogen
7. (英) Magnetic Resonance Spectroscopy
8. (英) Molecular Sequence Data
9. (英) NADP
10. (英) Pyrococcus horikoshii
11. (英) Recombinant Proteins
12. (英) Sequence Alignment
13. (英) Sequence Homology, Amino Acid
14. (英) Serine
15. (英) Stereoisomerism
16. (英) Substrate Specificity

1. (英) Article.Affiliation: Department of Biological Science and Technology, Faculty of Engineering, The University of Tokushima, 2-1 Minamijosanjimacho, Tokushima, Japan.

2. (英) Article.PublicationTypeList.PublicationType: Journal Article

3. (英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't