| Budget Amount *help |
¥55,640,000 (Direct Cost: ¥42,800,000、Indirect Cost: ¥12,840,000)
Fiscal Year 2014: ¥11,570,000 (Direct Cost: ¥8,900,000、Indirect Cost: ¥2,670,000)
Fiscal Year 2013: ¥12,220,000 (Direct Cost: ¥9,400,000、Indirect Cost: ¥2,820,000)
Fiscal Year 2012: ¥12,740,000 (Direct Cost: ¥9,800,000、Indirect Cost: ¥2,940,000)
Fiscal Year 2011: ¥12,610,000 (Direct Cost: ¥9,700,000、Indirect Cost: ¥2,910,000)
Fiscal Year 2010: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
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| Outline of Final Research Achievements |
We developed a self-excising Cre/loxP-mediated marker recycling system with mutated lox sequences to introduce a number of biosynthetic genes into Aspergillus oryzae. By using the marker recycling system constructed, hyperproduction of kojic acid could be achieved in A. oryzae by the introduction of the two genes that each encodes oxidoreductase and transporter. Deletion of the transporter resulted in a significant decrease in the yield of kojic acid compared to the wild type strain, indicating that overexpression of the transporter gene is highly effective in improving the production yield of secondary metabolites. In addition, expression vectors with multiple expression cassettes consisting of the amyB promoter and terminator were also constructed. We examined subcellular localization of the enzyme proteins encoded by a gene cluster involved in aphidicolin biosynthesis from Phoma betae, which were expressed as GFP-fusion proteins in A. oryzae.
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