Development of a thermostable firefly luciferase

Autor:	Christopher R. Lowe, M.J Murphy, James A. H. Murray, Peter John White, L. C. Tisi, D. J. Squirrell
Rok vydání:	2002
Předmět:	chemistry.chemical_classification Chemistry Point mutation Mutant Wild type Protein engineering Biochemistry Analytical Chemistry Enzyme Environmental Chemistry Luciferase Spectroscopy Thermostability
Zdroj:	Analytica Chimica Acta. 457:115-123
ISSN:	0003-2670
DOI:	10.1016/s0003-2670(01)01496-9
Popis:	Firefly luciferase forms the basis of a wide range of analytical techniques. However, the enzyme is unstable and rapidly loses activity even at room temperature. This leads to losses in sensitivity and precision in analytical applications and also severely limits the fieldability of devices incorporating luciferase-based technologies. A number of point mutations have previously been identified that significantly increase the thermostability of the enzyme. We show here that when such mutations are combined they can have an additive effect on the stabilisation of the enzyme. As such, we have constructed a luciferase mutant containing four point mutations, relative to the wildtype enzyme, resulting in remarkably greater thermostability.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_________::f1ef070f5e3adc8d527c18a83650350e https://doi.org/10.1016/s0003-2670(01)01496-9 Zobrazit plný text záznamu Full Text from ScienceDirect