An optimized luciferase bioluminescent assay for coenzyme A

• Type: Article in International Scientific Journal
• Date: 2008

Authors

marques, sm (Author) Other; da silva, jcge (Author) FCUP

Journal

Title: Analytical and Bioanalytical Chemistry

Vol. 391 No. 4

Pages: 2161-2168

ISSN: 1618-2642

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 14 Citations

Scopus - 16 Citations

Scientific classification

FOS: Natural sciences > Biological sciences

Other information

Authenticus ID: P-003-XVR

DOI: 10.1007/s00216-008-2117-6

Abstract (EN): A new bioluminescent method for coenzyme A (CoA) quantification is described. It is based on the enzymatic conversion of dehydroluciferyl-adenylate (L-AMP) into dehydroluciferyl-coenzyme A (L-CoA) by firefly luciferase (E.C. 1.13.12.7) (LUC), which causes a flash of light that can be measured in a luminometer. The method was subjected to optimization using experimental design methodologies to obtain optimum values for the concentrations of L-AMP ([L-AMP]), luciferase ([LUC]), ATP ([ATP]) and luciferin ([LH(2)]). This method has a linear response over the range of 0.25-4 mu M of CoA, with a limit of detection (LOD) of 0.24 mu M and a limit of quantification (LOQ) of 0.80 mu M. The assay has a relative standard deviation of about 7%. By coupling this optimized procedure to bioluminescent detection, a sensible and robust method can be obtained for the analysis of CoA.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 8