1997 Fiscal Year Final Research Report Summary
The key reaction for carbon dioxide fixation in reductive TCA cycle
| Project/Area Number |
08660094
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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 |
応用微生物学・応用生物化学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
ISHII Masaharu The University of Tokyo, Grad.Sch of Agei.Life Sci.Department of Biotechnology, Associate professor, 大学院・農学生命科学研究科, 助教授 (30193262)
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| Project Period (FY) |
1996 – 1997
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| Keywords | reductive TCA / antotroph / cycle |
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| Research Abstract |
In order to clarify that reductive TCA cycle is operative in a thermophilic hydrogenxidizing bacterium, Hydrogenobater thermophilus, the followings were demonstrated.
1.Purification and characterization of ferredoxin
Purification of ferredoxin was performed by applying cell-free extract successively. to Q-Sepharose, DEAE-5PW,and Superdex 75. Characterization of the prorein was also perfprmed.
2.Purification and characterization of pyruvate synthase
Purification of pyruvate synthase was performed by applying cell-free extract successively to DEAE-Sepharose CL-6B,PA-QA,Hydroxyapatite, and Superdex 200. Characterization of the protein was also performed.
3.Reactivity of ferredoxin toward 2-oxoglutarate synthase or pyruvate synthase
In the first step, reaction was performed toward CO_2 evolution direction ; both enzymes reacted with ferredoxin. In the next step, reduced ferredoxin was used as a reductant to show the carboxylation reaction. However, only a little carboxylation reaction occurred probably due to the instability of CoA-compounds. Finally, CoA-compounds were produced through enzymatic reactions ( ATP : citrate lyase and succinyl-CoA synthetase ) . By this system carboxylation reaction was detected and also the expected products were identified.
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