2003 Fiscal Year Final Research Report Summary
Metabolic flux analysis using isotope distribution together with gene and protein expression analyses
| Project/Area Number |
14350438
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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 | Kyushu Institute of Technology |
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Principal Investigator |
SHIMIZU Kazuyuki Kyushu Institute of Technology, Kyushu Institute of Technology, Dept.of Bioscience and Bioinformatics, Professor (00150318)
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| Project Period (FY) |
2002 – 2003
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| Keywords | Metbolic engineering / Protein expression / gene expression / Isotope distribution / NMR / GC-MS / Metabolic flux distribution / Metabolic regulation |
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| Research Abstract |
In order to develop a quantitative model for the main metabolic pathway of Escherichia coli, we cultivated E.coliK12 and W3110, as well as E.coli BW25113 developed in Keio University, and its several single-gene-knockout mutants (zwf^-, pykF^-, gnd^-, ppc^-, pck^-, pgi^- etc.) in batch and continuous modes. We also cultivated several gene knockout mutants (pflA, B, C, D-, pta-, ppc-, adhA- etc.) in microaerobic condition for lactate production. Then we made the metabolic flux analysis based on ^<13>C labeling experiments, measured enzyme activities and the intracellular metabolite concentrations as well as gene expressions using RT-PCR to understand the metabolic regulation quantitatively for the modeling.
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