| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2022: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2021: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
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| Outline of Final Research Achievements |
Intracellular metabolic pathways suitable for enzymatic cascade reactions were constructed. We proceeded to construct the vanillin production pathway as one of the models for the reaction. It diverges from dehydroshikimic acid through protocatechuic acid to vanillic acid and protocatechuic aldehyde, from which it converges to vanillin. To construct this model pathway, we introduced three genes, DSD, OMT, and ACAR, and evaluated intermediate and vanillin production. Vanillin production was confirmed by gene transfer, and the respective metabolites of the pathway, including intermediates, were shown to fluctuate according to the expression levels of each enzyme. By examining the culture conditions, such as carbon source, and reexamining the gene expression levels, we were able to construct microbial strains that produced the intermediates at various rates. From this, it can be said that the foundation of the coenzyme regeneration system using ammonia has been established.
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