Biosynthesis-inspired deracemizative production of firefly D-luciferin

2016 Fiscal Year Final Research Report

• Project/Area Number: 26410185
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Kagoshima University
• Principal Investigator: Kato Dai-ichiro 鹿児島大学, 理工学域理学系, 助教 (60423901)
• Co-Investigator(Renkei-kenkyūsha): NEGORO Seiji 兵庫県立大学, 大学院工学研究科, 教授 (90156159) ; TAKEO Masahiro 兵庫県立大学, 大学院工学研究科, 准教授 (40236443)
• Research Collaborator: NAKAJIMA Yoshihiro 国立研究開発法人産業技術総合研究所, 健康工学研究部門, 研究グループ長 ; SHIRAKAWA Daiki 兵庫県立大学, 大学院工学研究科, 学生 ; OKUDA Masatoshi 兵庫県立大学, 大学院工学研究科, 学生 ; MAEDA Juri 鹿児島大学, 大学院理工学研究科, 学生
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: バイオテクノロジー / ホタル / 生物発光 / 生合成経路 / トランスクリプトーム解析

Outline of Final Research Achievements

In this project, we tried to establish a chiral free bioluminescence system inspired by the biosynthetic pathway of firefly D-luciferin. In fireflies, L-luciferin is the biosynthetic precursor of D-luciferin, which is produced from the L-form undergoing deracemization. This deracemization consists of three successive reactions: L-enantioselective thioesterification by luciferase, in situ epimerization, and hydrolysis by thioesterase. At first, a deracemizative luminescence system using a combination of firefly luciferase and fatty acyl-CoA thioesterase from bacteria was constructed. We could confirm that this firefly luciferase-thioesterase combination system worked well and can improve bioanalysis applications using the firefly bioluminescence reaction by efficient deracemization of D-luciferin. We also performed a transcriptome analysis toward adult firefly.

Free Research Field

生物有機化学