| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
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| Outline of Final Research Achievements |
In the field of bio-productions, identification of a rate-limiting step in the metabolic pathways is one of the important issues for enhancing the productivity. Although the maximum enzyme activity (Vmax) provides a clue for the identification, measuring the Vmax values of many reactions at once is a time-consuming task. Here, I developed a high-throughput method for simultaneously estimating the Vmax of the enzymes in a pathway using a time-course data of intermediate concentrations obtained from an in vitro experiment. The Vmax values of reactions in glycolysis of Escherichia coli were estimated with this protocol. Metabolic control analysis using the estimated Vmax values revealed a rate-limiting step. The validity was confirmed by additional in vitro and in vivo experiments.
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