| Budget Amount *help |
¥15,990,000 (Direct Cost: ¥12,300,000、Indirect Cost: ¥3,690,000)
Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2015: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2014: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000)
|
|---|
| Outline of Final Research Achievements |
Amide/ester compounds have been synthesized by using the reverse reaction of hydrolases by limiting water content in the reaction mixtures. In thermodynamically controlled synthesis, the final yields are essentially determined by the water content within the solvent environment. The yields of amide-synthesis obtained by reverse reaction using 6-aminohexanoate-dimer hydrolase in 90% t-butyl alcohol were drastically varied following several amino acid substitutions located at the entrance of the catalytic cleft of the enzyme. A movement of a loop region and a flip-flop of Tyr170 in the enzyme generate the local hydrophobic environment at the catalytic center. Here, we propose that the shift of the internal equilibrium between enzyme/substrate complex and enzyme/product complex by the “water-excluding effect” alters the rate of the forward/reverse reactions, and that the local hydrophobic environment potentially provides a reaction center suitable for efficient amide synthesis.
|
