Purification and Characterization of Aminoglycoside-Resistant inhibitior from methylotrophic Actinomycetes

Methanol 자화 방선균으로부터 Aminoglycoside 내성 저해물질의 정제 및 특성

  • 김현수 (계명대학교 자연과학대학 미생물학과) ;
  • 신재욱 (일양약품 중앙연구소)
  • Published : 1999.06.01


Methylotrophic actinomycetes No. 155 produced an aminoglycoside antibiotics(AG)-resistant inhibitor. We have previously reported that the inhibitor shows strong inhibition to sisomicin-resistant strain. In order to understand the functions of inhibitor and sisomicin-resistance, characterizations and purification of inhibitor were investigated. Strain No. 155 was tentatively identified as Nocardiopsis sp. based on morphological and some physiological characteristics. In the antimicrobial activity test, the addition of inhibitor to sisomicin showed a reduction effect of MIC on the test strains such as Gram(+), Gram(-) bacteria and yeasts. The combination of the inhibitor and various antibiotics revealed synergistic against E. coli K-12 and B. subtilis PCI 219. The induced intracellular proteins from sisomicin-resistant strain exhibited the sisomicin inactivation by invitro test. And the induced intracellular proteins were inactivated by addition of the inhibitor. The inhibitor compound was purified by anion exchange chromatography(Dowex-1) and HPLC using Asahipak ES-502C column. The purified inhibitor compound was detected in a single peak(above 98.5% purity) through the HPLC analysis.


aminoglycoside antibiotics;sisomicin;methylotrophic actinomyceted;sisomicin-resistant inhibitor


  1. J. Antibiot. v.29 Mocardicin A, a new monocyclic ?-lactam antibiotic Aoki, H.;H.Sakai;M. Kohsaka;T. Konomi;J. Hosoda;Y. Kuboshi;E. Igushi;H. Imanaka
  2. Antimicrob. Agents Chemother v.33 Classification of ?-lactamase:Group 1, 2a, 2b, 2b' Bush, K.
  3. Antimicrob. Agents Chemother v.38 Streptomycin resistance in mycobacteria Honore, N.;S.T. Cole
  4. J. Antimicrob. Chemother v.18 Comparative activities of the ?-lactamase inhibitors YTR830, clavulanate and sulbactam combined woth extended spectrum penicillins against ticar-cillin-resistant Enterobacteriaceae and Pseudomonads Jacobs, M.R.;S.C. Aranoff;S. Johenning;S. Yamabe
  5. J. Clin. Microbiol. v.34 Comparison of rectal and perirectal seabs for detection of colonization. with vancomycin-resistanct Enterococci Weinstein, J.W.;S. Tallapragada;P. Farrel;L.M. Dembry
  6. Antimicrob. Agents Chemother v.41 Mechanism of sulfonamide resistance in clinical isolates of Strptococcus pneumoniae Jeffrey, P.M.;A.M. Sefton;L.M. Hall
  7. J. Antibiot. v.32 Olivanic acids, a family of ?-lactamase inhibitory properties produced by Streptomyces species Ⅱ. Isolation and characterization of the olivanic acid MM4550, MM13902 and MM17880 from Streptomyces olivaceus Hood, J. O;S.J. Box;M.S. Verrall
  8. Microbiol. Rev. v.57 Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes Shaw, K.J.;P.N. Rather;R.S. Hare;H. Miller
  9. J. Antibiot v.29 Naturally-occuring ?-lactamase inhibitors with antibacterial activity Brown, A.G.;D. Butterworth;G. Hanscomb;J.D. Hood;C. Reading;G.N. Rolinson
  10. J. Antimicrob. Chemother. v.17 Variation in the potentiation of ?-lactam antbiotic activity by clavulanic acid and sulbactam against multiply antibiotic resistant bacteria Aldridge, K.E.;C.V. Sanders;R.L. Marier
  11. Proc. Natl. Acad. Sci. v.70 Aminoglyside antibiotic-inactivating enzymes in actinomycetes similar to those present in clinical isolates of antibiotic-resistant bacteria Benveniste, T.;J. Davies
  12. Antimicrob. Agents Chemother v.35 Aminoglycoside resistance and aminoglycoside usage:Ten years of experience in one hospital Gerding, D.N.;T.A. Larson;R.A. Hughes;M. Welier;C. Shanholtzer;L.R. Peterson
  13. Antimicrob. Agents Chemother v.32 Resistance to ticarcillin-potassium clavulanate among clinical isolates of the family Enter-obacteriaceae:Role of PSE-1 ?-lactamase and high levels of TEM-1 and SHV-1 and problems with false susceptibility in disk diffusion tests Sanders, C.C.;J.P. Iaconis;G.P. Bodey;G. Samonis
  14. bioscience & Bioindustry v.52 The present state and furture of infection symptom in hospital from MRSA(methicillin-resistant Staphylococcus aureus) Hiramatsu.K.
  15. J. Infect. Dis. v.167 A ribosomal gene mutation in streptomycin-resistant Mycobactrium tuberculosis isolates Douglass, J.;L.M.Steyn
  16. N. Engl. J. Med. v.319 Plasmid-mediated resistance to vancomycin and teicoplanin in Enterococcus faesium Leclercq, R.;E. Derlot;J. Duval;P. Courvalin
  17. J. Clin. Microbio v.34 Use of inhouse studies of molecular epidemiology and full species identification for controlling spread of vancomycinresistant Enterococcus faecalis isolates Bodnar, U.R.;G.A. Noskin;T. Suriano;I. Cooper;B.E. Reisberg;L.R. Peterson
  18. Antimicrob. Agents Chemother v.34 ?-Lactamase production in members of the family Enterobacteriaceae and resistance to >-lactam-enzyme inhibitor conbinations Thomson, K.S.;D.A. Weber;C.C. Sanders;W. E. Sanders
  19. Bergey's Manual of Systematic Bacteriology v.4 Williams, S.T.;M.E. Sharpe;J.G. Holt
  20. Annula Rev. Microbiol. v.32 Plasmid-determenied resistance to antimicrobial agents Davis, J.;D.I. Smith
  21. Kor. J. Biotechnol. Bioeng. v.12 Production of aminoglycoside-resistant inhibitor from methylotrophic actinomycetes Kim, H.S.;J.O. Shin;T.S. Yu
  22. Biochem. J. v.251 Beta-lactamase inhibitors-the inhibition of serine beta-lactamase by specific boronic acid Crompton, L.E.;B.K. Cuthbert;G. Lowe;S.G. Waley
  23. Mol. Microbiol. v.9 Molecular basis of streptomycin resistance in Mycobacterium tuberculosis:Alterations of the ribosomal protein S12 gene and point mutations within a functional 16S ribosonal RNA pseudoknot Finken, M.;P. Kirschner;A. Wrede;E.C. Bottger
  24. J. Clin. Microbiol. v.34 Vancomycin-resistant Aureobacterium species cellulitis and bacteremia in a patient with acutemyelogenous leukemia Nolte, F.;K.E. Arnold;H. Sweat;E.F. Winton;G. Funke
  25. Anal. Biochem. v.72 A rapid and sensitive method for the quantitation of protein utilizing the principle of protein-dye binding Bradford, M.M.
  26. J. Antibiot. v.25 Studies on penicillinase inhibitors produced by microorgnism Hata, T.;S. Omura;Y. Iwai;H. Ohno;H. Takeshima;N. Yamaguchi
  27. J. Antimicrob. Chemother v.13 Clavulanic acid in combination with ticarcillin:An in vitro comparison with other ?-lactams Clarke, A.M.;S.J.V. Zemcov
  28. The Best Guidance of a Study of Antibiotics Ohno, M;S. Omura
  29. Antimicrob. Agents Chemother v.39 New mobile gene cassttes containing an aminoglycoside resistance gene, aacA7, and chloramphenicol resistance gene, catB3, in an integron on pBWH301 Bunny, K.L;R.H. Hall;H.W. Stokes
  30. Antimicrob. Agents Chemother v.41 Emergence of an inhibitor-resistant ?-lactamase (SHV-10) derived from an SHV-5 variant Efthimios, E.P.;V. Miriagou;E. Tzelepi;M. Gazouli;L.S. Tzouvelekis
  31. The Society for Actinomycetes Japan, Tokyo. Experimental Methods of Identification of Actinomycetes Hamada, M.;M. Manabe
  32. Biochem. J. v.251 ?-Lactam resistance mechanisms by specific boronic acid Cole, S.T.;M.H. Nicolas
  33. Antimicrob. Agents Chemother v.41 Identification and characterization of IS1476, an insertion sequence-like element that disrupt van Y function in a vancomycin-resistant Enterococcus faecium strain Martin, G.M.;M.A. Drebot;G.J. Tyrrell
  34. J. Bacteriol. v.171 Involvement of multiple genetic determinants in high-level methicillin resistance in Staphylococcus aureus Murakami, K.;A. Tomasz