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1 March 2003 Creation of a Thermostable Firefly Luciferase with pH-insensitive Luminescent Color
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Abstract

The thermal instability and pH-sensitive spectral property of firefly luciferase have hampered its use as a sensitive multicolor luminescent label or bioluminescent resonance energy transfer donor. With the intention of improving the thermostability of a previously found firefly Hotaria parvula luciferase mutant with minor pH-sensitive spectral change (V368A), further mutation (E356R) was introduced by taking a reportedly stabilized mutant of Photinus pyralis luciferase into account. The double mutant E356R/V368A showed significantly improved thermostability because >90% activity remained after incubation for 1 h at 45°C, with its specific activity being maintained. Unlike the wild type or V368A, E356R/V368A showed no change in the emission maximum of 568 nm even at pH 6.3, also implying a mutual relationship between thermostability and the proportion of yellow–green luminescent peak under acidic condition.

Atsushi Kitayama, Hiroaki Yoshizaki, Yoshihiro Ohmiya, Hiroshi Ueda, and Teruyuki Nagamune "Creation of a Thermostable Firefly Luciferase with pH-insensitive Luminescent Color," Photochemistry and Photobiology 77(3), 333-338, (1 March 2003). https://doi.org/10.1562/0031-8655(2003)077<0333:COATFL>2.0.CO;2
Received: 9 August 2002; Accepted: 1 December 2002; Published: 1 March 2003
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