| Project/Area Number |
07556048
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
General fisheries
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
SATO Yoshihiko Kyoto Univ., Agriculture, Associate Professor, 農学研究科, 助教授 (60153970)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MARUYAMA Tadashi Marine Biotechnology Inst., Senior Researcher, 主任研究員
ISHINO Yoshizumi Biomolecular Engineering Res.Inst., Molecular Biology, Senior Researcher, 生物分子工学研究所, 主任研究員
UCHIDA Aritsune Kyoto Univ., Agriculture, Professor, 農学研究科, 教授 (50027190)
石田 祐三郎 京都大学, 農学部, 教授 (20026488)
|
|---|
| Project Period (FY) |
1995 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1996: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥3,100,000 (Direct Cost: ¥3,100,000)
|
|---|
| Keywords | Hyperthermophiles / Archaea / Hydrothermal vent / Heat stable enzyme / CO_2 fix / protease / DNA polymerase / CO_2固定 / PCR |
|---|
| Research Abstract |
In this study, we firstly isolate the useful marine hyperthermohpilic bacteria for genetic engineering, medicine, food science and chemical engineering. Secondly we purify heat and chemical stable enzymes and clone their genes for the development of unknown genetic resources.
The results are as follows ;
(1)New anaerobic hyperthermophilic archaeon Pyrococcus sp.growing optimally at 100ﾟC was isolated from a shallow marine hydrothermal vent in Tachibana Bay. The gene for DNA polymerase of this archaeon was cloned and expressed in E.coli. The amino acid sequence of this gene was closely relate to that of pol A from P.furiosus.
(2)New aerobic thermophilic bacterium Rhodothermus obamensis OKD7 growing optimally at 80ﾟC was isolated from the same station. CO_2 fixiing enzyme (PEPC) was purified and characterized. The allosteric enzyme PEPC was a homotetramer of 400kDa and showed maximum activity at 75ﾟC.The cloned gene of PEPC was expressed in E.coli. A thermostable recombinant PEPC showed similar property of the wild type.
(3)A novel aerobic hyperthermophilic archaeon Aeropyrum pernix K1 growing optimally at 90ﾟC was isolated from a costal soltataric vent at Kodakara Island. Strain K1 produced an extracellular metalloprotease with a molecular mass of 52kDa. The optimum temperature for the enzyme was 100ﾟC.The enzyme was extremely thermostable showing half-life of 1.2h at 125ﾟC.
|
