| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2017: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2016: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
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| Outline of Final Research Achievements |
To practical alkane production in E.coli cells, we need to explore alkane synthesis enzymes, AAR and ADO, with higher activity, engineer a pathway to enhance a substrate (acyl-ACP) production and delete genes competing alkane production. At first, we compared catalytic activity of AAR and ADO derived from various cyanobacteria and successfully identified the highest active AAR and ADO. In parallel, we genetically engineered E.coli cells to emit bioluminescence depending on yield of aldehyde production in the cells. Using this engineered cells and a high-sensitive chemi-luminescence imager, we can evaluate amounts of aldehyde production in cells in real-time by monitoring bio-luminescent intensities emitted from over 1000 colonies on agar plate. This method enables high through-put screening to find gene disrupted strains, which enhanced acyl-ACP production, and/or decreased aldehyde consumption in competitive enzyme reaction, among library of strains deleted genes randomly.
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