Functional expression of RubisCO by a high-speed carbon dioxide fixing microorganism
| Project/Area Number |
07455325
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Shinshu University (1996)
The University of Tokyo (1995) |
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Principal Investigator |
KODAMA Tohru Shinshu University, Faculty of Textile Science and Technology, Professor, 繊維学部, 教授 (30011901)
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| Co-Investigator(Kenkyū-buntansha) |
ISHII Masaharu The university of Tokyo, Department of Biotechnology, Assistant Professor, 大学院・農学生命科学研究科, 助手 (30193262)
YAMAKAWA Takashi The university of Tokyo, Department of Biotechnology, Assistant Professor, 大学院・農学生命科学研究科, 助手 (20134520)
IGARASHI Yasuo The university of Tokyo, Department of Biotechnology, Professor, 大学院・農学生命科学研究科, 教授 (90114363)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥6,500,000 (Direct Cost: ¥6,500,000)
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| Keywords | Rubis CO / CO_2 fixation / イオンクロマトグラフ / specificity factor / 活性化 |
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| Research Abstract |
A thermophilic hydrogen bacterium Pseudomonas hydrogenothermophila strain TH-1 possesses cbbQ gene under RubisCO gene (cbbLS). CbbQ protein was thought to participate in the post-translational activation/stabilization of RubisCO.
We showed the presence of cbbO gene under cbbQ gene and clarified the gene sequence. In the next step, plasmids harboring cbbQ,cbbO,cbbQ-cbbO gene were prepared. By using one of the above plasmids and a plasmid harboring cbbLS,transformation of E.coli was performed. Then, difference in activity or expression of RubisCO was analyzed by measuring RubisCO activity or by electrophoreis, respectively. When only the plasmid harboring cbbLS was used, RubisCO activity was low and the RubisCO protein was detected as a smear band on polyacrylamide-gel electrophoresis. Contrary to this, in the presence of cbbQ or cbbO gene, the activity of RubisCO increased 2 to 3 fold, also the RubisCO protein was detected as a clear band on polyacrylamide-gel electrophoresis.
In a separate experiments, three kinds of protein (CbbLS,CbbQ,and CbbO) were prepared for the experiment of in vitro RubisCO activation. However, successful results have not been obtained.
During our research, CbbO protein itself was proved to possess CO_2 incorporation activity, which will be a source of thinking for future research.
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Report
(3 results)
Research Products
(2 results)
