title: Establishment of an effective TLC bioautographic method for the detection of Mycobacterium tuberculosis H37Ra phosphoglucose isomerase inhibition by phosphoenolpyruvate.
abstract: A bioautographic assay based on thin layer chromatography was developed for phosphoenolpyruvate (PEP) detecting as a known but rarely studied inhibitor of phosphoglucose isomerase (PGI). The protocol with NADP(+)/NBT/PMS (β-nicotinamide adenine dinucleotide phosphate/nitrotetrazolium blue chloride/phenazine methosulfate) staining was capable of detecting Mycobacterium tuberculosis H37Ra PGI inhibition using PEP. According to this method, visibly brighter spots (zones) against purple background are observed in the area of inhibition of the above-mentioned enzyme activity. The detection limit for PEP as an inhibitor of Mycobacterium tuberculosis H37Ra PGI was 226 μg per spot/zone. Noteworthy is that we are the first authors to have successfully used a bioautographic assay to detect Mycobacterium tuberculosis H37Ra PGI inhibition by PEP.
authors.affiliation: a Chair and Department of Biochemistry and Biotechnology, Medical University of Lublin , Lublin , Poland and.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Paradowska
authors.affiliation: b Department of Physical Chemistry , Medical University of Lublin , Lublin , Poland.
authors.forename: Beata
authors.initials: B
authors.lastname: Polak
authors.affiliation: b Department of Physical Chemistry , Medical University of Lublin , Lublin , Poland.
authors.forename: Adam
authors.initials: A
authors.lastname: Chomicki
authors.affiliation: a Chair and Department of Biochemistry and Biotechnology, Medical University of Lublin , Lublin , Poland and.
authors.forename: Grażyna
authors.initials: G
authors.lastname: Ginalska
mesh_terms: D002855:Chromatography, Thin Layer / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005956:Glucose-6-Phosphate Isomerase / Q000037:antagonists & inhibitors*; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D010728:Phosphoenolpyruvate / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D010728:Phosphoenolpyruvate; D005956:Glucose-6-Phosphate Isomerase
keywords: Antimycobacterial activity; autography; glycolysis; mycobacterial cell wall; potential drug target