Creation of peptide ligase that enables protein-linked structure formation by novel function screening method

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K13313
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Nanobioscience
• Research Institution: The University of Tokyo
• Principal Investigator: Ichiki Takanori 東京大学, 大学院工学系研究科(工学部), 教授 (20277362)
• Co-Investigator(Renkei-kenkyūsha): UENO Shingo 川崎市産業振興財団, ナノ医療イノベーションセンター, 副主幹研究員 (30594650)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: ペプチドリガーゼ / タンパク質アレイ / 酵素改変 / Sortase

Outline of Final Research Achievements

It is expected to develop a technique for artificially designing and arranging various proteins responsible for biochemical reactions and reconstructing the biochemical reaction network.In this study, we propose a method to create peptide ligase that enables intermolecular coupling of proteins and peptides, and accumulation on solid phase, and developed and verified necessary basic technologies. We developed a technique to immobilize the cell-free synthesized protein on the solid phase using sortase, a protein transferase derived from gram-positive bacteria. Then we experimentally clarified the problems which become prominent in the coexistence of cell-free translation and sortase reaction system, and discussed their solutions.These results are expected to contribute to the future development of screening technology for sortase variants by combining with microarray chip technology.

Free Research Field

ナノバイオテクノロジー