Phosphoglucose isomerase from Escherichia coli ATCC 25922 – pilot studies
DOI:
https://doi.org/10.12923/Keywords:
phosphoglucose isomerase, isoenzymes, PGI inhibitors, Escherichia coli ATCC 25922, biofilmAbstract
Phosphoglucose isomerase (EC 5.3.1.9; PGI) is an enzyme participating in the metabolism of the sugar and carrying the reversible isomerization of glucose-6-phosphate and D-fructose-6-phosphate. It plays key role in the process of the extracellular polysaccharides (EPS) synthesis in vivo – creating the bacterial biofilm. The complete understanding of PGI catalysis and inhibition mechanisms may results in discovery of new PGI inhibitors – antibacterial drugs.
In this pilot study we described the procedure of partial purification of E. coli ATCC 25922 PGI from cytosolic fraction. The activity of enzymes was measured by the conversion of D-fructose-6P into D-glucose-6P. Two forms (isoenzymes I and II) of this enzyme were separated. Some properties of dominant form (isoenzyme I) of PGI were studied. The Km constant for D-fructose-6P was 1 mM. The enzyme had optimum activity in TRIS/HCl buffer, pH 7.65. Phosphoglucose isomerase had molecular weight 120 kDa. Our results indicated that amino acids - cysteine, tryptophan, tyrosine and lysine residues were to be presented in the active site of PGI from E. coli ATCC 25922. The lack of the activity inhibition of characterized enzyme by chelators as EDTA, 1,10-phenantroline and TRIS indicated that the PGI is not a metallprotein. A large similarity between properties of phosphoglucose isomerases isolated from E. coli ATCC 25922 and other E. coli strains such as K10 and ATCC 35218 was discovered.
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