α-Phosphoglucomutase from Escherichia coli ATCC 25922 – pilot studies
DOI:
https://doi.org/10.12923/Keywords:
α-phosphoglucomutase, Escherichia coli ATCC 25922, biofilmAbstract
The a-phosphoglucomutase (a-PGM, EC 5.4.2.2) is an enzyme participating in the metabolism of the sugar and carrying the reversible isomerization of a-D-glucose-1-phosphate to a-D-glucose-6-phosphate.
Some properties of a-PGM from E. coli ATCC 25922 were studied. The Km constant for a-D-glucose-1-phosphate was 25 × 10-5 M/L. The enzyme had optimum activity in TRIS/HCl buffer, pH 8.0. Its activity was stimulated by addition of a-D-glucose-1,6-bisphosphate, Mg(II) cations and several thiols. The a-phosphoglucomutase had molecular weight 65 kDa and consisted of one polypeptide chain. Our results indicated the significant sensitivity of enzymatic activity by non- and organic mercuric compounds. The lack of the activity inhibition of characterized enzyme by chelators as 2,4’-bipyridyl, EDTA and TRIS indicated that the a-PGM is not a metallprotein. The similarities and differences between properties of a-phosphoglucomutases isolated from E. coli ATCC 25922 and other E. coli strains such as ATCC 26 and ATCC 35218 were discovered.
Our pilot studies have an important implication in better understanding of virulence of enteric bacteria (E. coli).
References
1. Adhya S., Schwartz M.: Phosphoglucomutase mutants of Escherichia coli K-10. J. Bacteriol., 108, 621, 1971.
2. Belocopitow E., Marechal L.R.: Metabolism of trehalose in Euglena gracilis. Eur. J. Biochem., 46, 631, 1974.
3. Bradford M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of the protein dye binding. Anal. Biochem., 72, 248, 1976.
4. Brautaset T., Standal R., Fjærvik E. et al.: Nucleotide sequence and expression analysis of the Acetobacter xylinum phosphoglucomutase gene. Microbiol., 140, 1183, 1994.
5. Buchanan J.T., Stannard J.A., Lauth X. et al.: Streptococcus iniae phosphoglucomutase is a virulence factor and a target for vaccine development. Infect. Immun., 73, 6935, 2005.
6. Davis B.: Disc electrophoresis. II Method and application to human serum proteins. Ann. N.Y. Acad. Sci., 121, 404, 1964.
7. Degeest B., de Vuyst L.: Correlation of activities of the enzymes α-phosphoglucomutase, UDP-galactose 4-epimerase, and UDP-glucose pyrophosphorylase with exopoly-saccharide biosynthesis by Streptococcus thermophilus LY03. Appl. Environ. Microbiol., 66, 3519, 2000.
8. Dworniczak K., Ginalska G., Patkowska M.: Isolation and characterization of phophoglucomutase (PGM) from Escherichia coli bacteria cells. Ann. UMCS., Sect. DDD, 99, 2008.
9. Hardy G.G., Magee A.D., Ventura C.L. et al.: Essential role for cellular phophoglucomutase in virulence of type 3 Streptococcus pneumoniae. Infect. Immun., 69, 2309, 2001.
10. Ishaque A., Milhausen M., Levy H.R.: On the absence of cysteine in glucose 6-phosphate dehydrogenase from Leuconostoc mesenteroides. Biochim. Biophys. Res. Commun., 59, 894, 1974.
11. Joshi J.G., Handler P.: Phoshoglucomutase. I. Purification and properties of phosphoglucomutase from Escherichia coli. J. Biol. Chem., 239, 2741, 1964.
12. Kvam C., Olsvik E.S., McKinley-McKee J. et al.: Studies on recombinant Acetobacter xylinum -phosphoglucomutase. Biochem. J., 326, 197, 1997.
13. Laemmli U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680, 1970.
14. Lu M., Kleckner N.: Molecular cloning and characterization o the pgm gene encoding phosphoglucomutase of Escherichia coli. J. Bacteriol., 176, 5847, 1994.
15. Marechal L.R. , Oliver G., Veiga L.A. et al.: Partial purification of some properties of beta-phosphoglucomutase from Lactobacillus brevis. Arch. Biochem. Biophys., 228, 592, 1984.
16. Mehra-Chaudhary R., Mick J., Tanner J.J. et al.: Crystal structure of a bacterial phosphoglucomutase, an enzyme involved in the virulence of multiple human pathogens. Proteins, 79, 1215, 2011.
17. Qian N. , Stanley G.A., Hahn-Hägerdal B. al.: Purification and characterization of two phosphoglucomutases from Lactococcus lactis susp. lactis and their regulation in maltose- and glucose-utilizing cells. J. Bacteriol, 176, 5304, 1994.
18. Schackelford G.S., Regni C.A., Beamer L.J.: Evolutionary trace analysis of the -D-phosphoxemutase superfamily. Protein Sci., 13, 2130, 2004.
19. Sjöberg A., Hahn-Hägerdal B.: β-glucose-1-phosphate, a possible mediator for polysaccharide formation in maltose-assimilating Lactococcus lactis. Appl. Environ. Microbiol., 55, 1549, 1989.
20. Szynal K.: Purification and evaluation of biological role of phosphoglucomutase (PGM) and phosphoglucose isomerase (PGI) from bacteria cells of Escherichia coli. Master’s thesis. Medical University of Lublin, 2008.
21. Yamane K., Ogawa K., Yoshida M. et al: Identification and characterized of clinically isolated biofilm-forming Gram-positive rods from teeth associated with persistent apical periodontitis. J. Endod., 35, 347, 2009.
Downloads
Published
Issue
Section
License
Copyright (c) 2011 Authors
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 Unported License.
