| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
|
|---|
| Outline of Final Research Achievements |
It is too low for activities of enzymes (AAR and AD), to produce alkane by microorganisms, practically. Therefore. in this study, we proposed that the enzyme activities were improved by directed evolution. Directed evolution is a method to improve and/or change characteristics and activities of enzymes, and require a high-throughtput method to evaluate AD and AAR activities of mutants, quickly and simply.
At first, using Nile red, we tried to detect amount of alkane produced by co-expression of AD and AAR as s in vivo AD activity, however, we did not discriminate activities among AD mutants. Thus, we needed to change a strategy to elucidate in vivo AD activity. On the other hand, we successfully established a method to evaluate in vivo AAR activity directly as bioluminescence by coexpressed bacterial luciferase. Using this method, we can simultaneously compare in vivo AAR activity among over 1000 AAR mutants, resulting in accelerating improvement of AAR activity by directed evolution.
|
