Activity coupling of Vibrio harveyi luciferase and flavin reductase (FRP): Oxygen as a probe

作者：Li Xi; Tu Shiao Chun^*

来源：Archives of Biochemistry and Biophysics, 2006, 454(1): 26-31.

DOI：10.1016/j.abb.2006.07.016

• channeling
• reduced flavin
• monooxygenase
• two-component
• luciferase
• bacterial
• flavin reductase
• direct transfer
• flavin
• diffusion
• oxygen variation
• kinetics of coupled reaction
• bioluminescence
• bacterial

Several lines of evidence have been reported previously to document the ability of the Vibrio harveyi NADPH-specific flavin reductase FRP to directly transfer reduced riboflavin-5'-phosphate to luciferase for bioluminescence. This study aimed at characterizing further the kinetic properties of FRP in such a direct channeling system and investigating whether the complete direct transfer of reduced flavin was the exclusive pathway in the FRP:luciferase coupled bioluminescence reaction. To these ends, a new kinetic approach of oxygen variation was employed. Results indicated that increases in oxygen concentration led to gradual decreases of the peak bioluminescence intensity, K-m,K-FMN, and K-m,K-NADPH of FRP in the coupled reaction. In comparison with theoretical schemes, these findings indicated that the FRP:luciferase coupled reaction can utilize reduced flavin by both free diffusion and direct transfer. The upper limits of the true K-m,K-FMN and K-m,K- NADPH of FRP in the direct transfer system were. determined.

• 出版日期2006-10-1