| Outline of Final Research Achievements |
In microbial production of industrial chemicals, the export of products to extracellular space is one of the rate-limiting factors. In this study, to achieve efficient production of membrane-impermeable organic acids, we performed thermodynamic analysis and structural analysis of the organic acid efflux transporter. First, we tried to determine the thermodynamic parameters by using isothermal titration calorimetry, and succeeded in measuring the heat generated during transporter-substrate binding. In addition, to clarify the molecular structure of organic acid efflux transporter by X-ray crystallography, we carried out crystallization of the purified protein and X-ray diffraction experiments. Single particle analysis using cryo-electron microscopy was also carried out.
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