Construction of bilin reductase library, promotes next-generation optogenetics.

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K05758
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: Kurume National College of Technology
• Principal Investigator: HAGIWARA Yoshinori 久留米工業高等専門学校, 生物応用化学科, 准教授 (10628548)
• Co-Investigator(Kenkyū-buntansha): 杉島 正一 久留米大学, 医学部, 准教授 (30379292)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: 光合成色素 / ビリン色素 / オプトジェネティクス / 光遺伝学

Outline of Final Research Achievements

The purpose of this research is to construct an enzyme library that produces bilin pigments accepting each light color. By functionally designing of bilin with enzymatic modification, we combine their bilins with photosensor proteins and aimed to promote optogenetics for multi-switch control using arbitrary light colors.
By adjusting the concentration ratio of the protein group that supplies electrons to the enzyme and proceeding with the enzymatic reaction, not only the synthesis rate of the final product but also the synthesis rate and decomposition rate of the intermediate product in the enzymatic reaction can be evaluated stability. The pigments considered to be final products and intermediate products were analyzed by HPLC. Site-directed mutagenesis was also investigated to increase the yield of intermediate as billin.

Free Research Field

構造生物学

Academic Significance and Societal Importance of the Research Achievements

ビリン色素はヘムの代謝産物であるため、植物のほか、動物や細菌に至るまで存在する色素である。2017年には、新生児黄疸の原因ともなるビリルビンを合成する酵素の結晶構造が、本研究者が参画した研究によって明らかとなり、ビリン色素の基礎医学的側面からの応用にも光が当てられている (H. Takao, et al., Nat. Commun., 2017)。
生物界に広く存在するビリン色素の改変を目指す本研究によって、有機化学合成が困難なビリン色素を酵素化学的に自在にデザインすることで、光センサータンパク質が鍵を握るオプティジェネティクスの新たな展開が期待できる。