2007 Fiscal Year Final Research Report Summary
Application of Hyperthennophilir 2-Deoxy-D-Riltose-5-Phosphate Aldolase
| Project/Area Number |
17613002
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
極限環境生物学
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| Research Institution | The University of Tokushima |
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Principal Investigator |
SAKURABA Haruhiko The University of Tokushima, Institute of Technology and Science, Associate Professor (90205823)
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| Co-Investigator(Kenkyū-buntansha) |
OHSHIMA Toshihisa Kyushu University, Institute of Genetic Resources, Faculty of Agriculture, Professor (10093345)
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| Project Period (FY) |
2005 – 2007
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| Keywords | DERA / hyperthermophile / Pyrobaculum aerophilum / Thermotoga maritime / archaea / sequential aldol condensation / acetaldehyde / 2-deoxy-D-ribose-5-phosphate aldolase |
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| Research Abstract |
Genes encoding 2-deoxy-D-ribose-5-phosphate aldolase (DERA) homologues from two hyperthermophiles, the archaeon Pyrobaculum aerophilum and the bacterium Thermotoga maritima, were expressed individually Eschelichia coli, after which the structures and activities of the enzymes produced were characterized and compared with those of E. coli DERA. lb our surprise, the two hyperthermophilic DERAs showed much greater catalysis of sequential aldol condensation using three acetaldehydes as substrates than the E coli enzyme, even at a low temperature (25℃), though both showed much less 2-deoxy-D-ribose-5-phosphate synthetic activity. Both the enzymes were highly resistant to high concentration of acetaldehyde and retained about 50% of their initial activities after 20-hours exposure to 300 mM acetaldehyde at 25℃, whereas the E coli DERA is almost completely inactivated after 2-hours exposure under the same conditions.
The structure of the P. aerophilum DERA was determined by x-ray crystallography to a resolution of 2.0A. The mainchain coordinate of the P. aerophilum enzyme monomer was quite similar to those of the T maritime and E call enzymes, whose crystal structures have been already solved. However, the quaternary structure of the hyperthermophilic enzymes was totally different from that of the E coli DERA. The area of the subunit-subunit interface in the dimer of the hyperthermophilic enzymes are much larger compared with the E coif enzyme. This promotes the formation of the unique dimeric structure and strengthens the hydrophobic inter subunit interactions. These structural features are considered to be responsible for the extremely high stability of the hyperthermophilic DERAs.
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Research Products
(19 results)
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[Journal Article] Structure of L-aspartate oxidase from the hyperthermophilic archaeon Sulfolobus tokodaii2008
Author(s)
Sakuraba, H., Yoneda, K., Asai, I., Tsuge, H., Katunuma, N., Ohshima, T
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Journal Title
Biochim Biophys Acta 1784
Pages: 563-571
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Sequential aldol condensation catalyzed by hyperthermophilic 2-deoxy-D-ribose-5-phosphate aldolase2007
Author(s)
Sakuraba, H., Yoneda, K., Yoshihara, K., Satoh, K., Kawakami, R., Uto, Y., Tsuge, H., Takahashi, K., Hori, H., Ohshima, T
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Journal Title
Appl. Environ. Microbiol 73
Pages: 7427-7434
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Gene expression and characterization of 2-keto-3-deoxygluconate kinase, a key enzyme in the modified Entner-Doudoroff pathway of the aerobic and acidphilic hyperthermophile Sulfolobus tokodaii2007
Author(s)
Ohshima, T., Kawakami, R., Kanai, Y., Goda, S., Sakuraba, H
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Journal Title
Prot. Exp. and Purification 54
Pages: 73-78
Description
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