Principles for assembling cascade enzymes

2016 Fiscal Year Final Research Report

• Project/Area Number: 25248038
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Kyoto University
• Principal Investigator: MORII Takashi 京都大学, エネルギー理工学研究所, 教授 (90222348)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Keywords: 核酸関連化学 / タンパク質 / 単分子配置 / 酵素 / 代謝 / DNAナノテクノロジー / 受容体 / 人工代謝系

Outline of Final Research Achievements

Understanding the effect of spatial organization on the efficiency of multi-enzyme systems reveals a principle that efficiently drives multi-enzymatic cascade. DNA nanostructures provide useful scaffolds for the spatial organization of enzymes to form artificial enzyme cascades. We have developed methods to use sequence-specific DNA binding proteins as adaptors to stably locate the enzymes at specific positions on DNA scaffold. Our protein-adaptor-based method successfully assembled recombinant enzymes in high loading yields with distinctly controlling the number of enzyme molecules and maintaining the catalytic activities of enzymes. An artificial enzyme cascade of xylose reductase (XR) and xylitol dehydrogenase (XDH) has been constructed by coassembling the enzymes on DNA origami. Further extension of this artificial metabolic pathway was achieved by using modular adaptors equipped with the protein-tag to successfully construct XR/XDH/xylulose kinase.

Free Research Field

生物機能化学