The Purfication and Characterization of Macrolide-Phosphotransferase K of Escherichia coli 209K Highly Resistant to Erythromycin

에리스로마이신 고도내성 대장균 209K 유래 마크로라이드-포스포트란스페라제 K의 정제 및 특성

  • Published : 1997.06.01

Abstract

Resistance gene mphK was cloned from Escherichia coli 209K strain which is highly resistant to erythromycin (EM). By using the cloned plasmid pGE64, E. coli NM522 was transformed. The comparison of macrolide-phosphotransferase K [MPH(K)] activity between E. coli 209K and E. coli NM522(pGE64) showed that the total enzyme activity of MN522(pGE64) was fifty-fole higher than that of 209K. To identify characteristics of MPH(K) more precisely. MPH(K) was isolated and purified from the NM522 (pGE64). The final purification f MPH(K) through several stages of purification process was 89 fole and the overall recovery was 11%. This enzyme was monomer with the molecular weight of 34 kDa and its isoelectric point (pI) was 5.0. The optimal pH and temperature for activity were 8.0 and $40^{\circ}C$, respectively.

Keywords

References

  1. J. Antibiot. (A) v.19 Effect of antibiotics on growth of Mycoplasma pneumoniae Arai, S.;Yoshida, K.;Izawa, A.;Kumagai, K.;Ishida, N.
  2. J. Infect v.2 Comparison of erythromycin and oxytetracycline in the treatment of cervical infection by Chlamydia trachomatics Oriel, J. D.;Ridgway, G. L.
  3. American J. Med. v.69 Legionnaires disease Weissenburger, D. D.;Rappaport, N.;Ahlkwalia, M. S.;Melvani, R.;Renner, E. D.
  4. Antimicrob. Agents Chemother v.20 Vitro susceptibility of Campylobacter fetus subsp. jejuni to N-formimidoyl thienamycin. resamicin, cefoperazine, and other antimicrobial agents Ahonkhai, V.;Cherubin, C. E.;Sierra, M. F.;Bokkenheuser, V. D.;Shulman, M. A.;Mosenthal, A. C.
  5. Infection v.6 Effect of erythromycin and amoxycillin on Bordetella pertussis in the nasopharynx. Trollfors, B.
  6. J. Bacteriol v.150 Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide. lincosamide, and streptogramin type B antibiotics Horinouchi, S.;Weisblum, B.
  7. J. Bacteriol v.162 Nucleotide sequence of ermA, a macrolide-lincosamide-strepto-gramin B determinant in Staphylococcus aureus Murphy, E.
  8. Antimicrob. Agents Chemother. v.30 Contribution of two different mechanisms to erythromycin resistance in Escherichia coli Arther, M.;Courvalin, P.
  9. J. Antibiot v.41 Structure of a phosphorylated derivative of oleandomycin obtained by reaction of oleandomycin with an extract of an erythromycin resistant strain of Escherichia coli O'Hara, K.;Kanda, T.;Kono, M.
  10. Antimicrob. Agents Chemother. v.34 Role of an energy-dependent efflux pump in plasmid pNE24 mediated resistance to 14- and 15-membered macrolides in Staphylococcus epidermidis Goldman, R. C.;Capobianco, J. O.
  11. Nature v.196 Susceptibility of Proteus mirabilis and its stable L-forms to erythromycin and other macrolides Taubeneck, U.
  12. J. Antibiot v.37 Enzymatic hydrolysis of erythromycin by a strain of E. coli: a new mechanism of resistance Barthelemy, P.;Autissier, D.;Gerbaud, G.;Courvalin, P.
  13. Antimicrob. Agents Chemother. v.29 Epidemiology of intestinal colonization by members of the family Enterobacteriaceae highly resistant to erythromycin in a hematology oncology unit Andremont, A.;Sancho-Garnier, H.;Tancrede, C.
  14. Mol. Cells v.6 Nucleotide sequence, expression and transcriptional analysis of the Escherichia coli mphK gene encoding macrolide-phosphotransferase K. Kim, S. K.;Baek, M. C.;Choi, S. S.;Kim, B. K.;Choi, E. C.
  15. Gene v.96 High efficiency transformation of Escherichia coli with plasmids Inoue, H.;Nojima, H.;Okayama, H.
  16. Chemotheraphy v.34 Simple assay for determining streptomycin inactivation with intact cells of bacteria and its applications to P. aeruginosa clinically isolated Kono, M.;O'Hara, K.;Sato, K.;Ohmiya, K.
  17. J. Biol. Chem. v.193 Protein measurement with the Folin phenol reagent Lowry, O. H.;Rosebrough, N. J.;Farr, A. L.;Randall, R. J.
  18. Anal. Biochem. v.167 Rapid isoelectric focusing in a vertical polyacrylamide minigel system Robertson, E. F.;Dannelly, H. K.;Malloy, P. J.;reeves, H. C.