Cloning, expression and characterization of polyketide synthase gene from Scopolia Japonica

2016 Fiscal Year Final Research Report

• Project/Area Number: 26870425
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Natural medicines; Physical pharmacy
• Research Institution: Kyushu University
• Principal Investigator: Tabata Kaori (佐々木香織) 九州大学, 薬学研究院, 助教 (90464388)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: ポリケタイド合成酵素 / X線結晶構造解析

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.

Free Research Field

タンパク質科学