| Budget Amount *help |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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| Outline of Final Research Achievements |
Recent advances in protein chemistry have revealed that global conformational change of an enzyme is closely related to formation of the transition state and thus constitutes the rate-determining step in an enzyme reaction. Here, using two enzymes, acyl-ACP reductase (AAR) and aldehyde-deformylating oxygenase (ADO), which are indispensable for biofuel production, we detected conformational fluctuations of enzymes by molecular dynamics simulations (theory) and NMR relaxation measurements (experiment). We then searched for residues essential for controlling conformational fluctuations of the enzymes. Introduction of mutations at these residues resulted in changes of activities, indicating that regulation of the transition state of an enzyme is possible by such mutations. We also performed comprehensive mutational analysis including alanine-scanning mutagenesis on AAR and ADO by making more than 600 mutants and succeeded in identifying the residues important for controlling activities.
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