| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
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| Outline of Final Research Achievements |
Various important pharmacological compounds such as atropine and scopolamine are included in Scopolia japonica. These compounds are synthesized using ornithine as a starting material, however, it’s unclear which enzyme catalyzes the tropinone synthesis reaction. Hence, in the present study we aimed to identify function and structure of novel polyketide synthase (PKS) from S. japonica.
Two novel genes of PKS from S. japonica were obtained. The recombinant PKSs expressed in E. coli as a soluble fraction was highly purified using affinity and ion exchange column and the enzyme assay with HPLC showed that new compounds were synthesized using some substrates. Moreover, crystallization screening resulted in obtaining needle or plate crystals. We could get single crystals in an optimized condition. The crystal diffracted to 2.0 angstrom at KEK-PF and the structures of PKS were determined using molecular replacement method.
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