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Purification and characterization of inducible beta-lactamases in Aeromonas spp.

Purification and characterization of inducible beta-lactamases in Aeromonas spp. J P Iaconis and C C Sanders Department of Medical Microbiology, Creighton University School of Medicine, Omaha, Nebraska 68178. ABSTRACT beta-Lactamases from Aeromonas hydrophila and A. sobria were purified and characterized. Both species produced beta-lactamases that were inducible by either cefoxitin or imipenem. These species were resistant to ampicillin and cephalothin but not imipenem. Isoelectric focusing of sonic extracts revealed one band at pI 8.0 and a second band at pI 7.0 for A. hydrophila. Likewise, A. sobria produced two bands, one at pI 8.4 and the other at pI 7.0. Two enzymes from each species were separated by flatbed electrofocusing gel and purified to homogeneity. The molecular weight of the pI 7.0 enzyme (A1) from both species was estimated to be 42,500, whereas the pI 8.0 (A2h) and 8.4 (A2s) enzymes of A. hydrophila and A. sobria had molecular weights of 31,500 and 35,000, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The relative Vmax values for cephalothin, penicillin, and imipenem for these enzymes indicated that A1 was primarily a cephalosporinase while A2h and A2s were penicillinases highly active against carbapenems. A1 was susceptible to inhibition by cloxacillin, while the A2 enzymes were inhibited by clavulanic acid and EDTA and required zinc for activity. Thus, there appear to be two distinct inducible beta-lactamases in A. hydrophila and A. sobria that play an important role in the beta-lactam resistance of these species. CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? « Previous | Next Article » Table of Contents This Article doi: 10.1128/​AAC.34.1.44 Antimicrob. Agents Chemother. January 1990 vol. 34 no. 1 44-51 » Abstract PDF Classifications Research Article Services Email this article to a colleague Similar articles in ASM journals Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of AAC Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Iaconis, J. P. Articles by Sanders, C. C. Search for related content PubMed PubMed citation Articles by Iaconis, J. P. Articles by Sanders, C. C. Related Content Load related web page information Social Bookmarking CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? current issue December 2011, volume 55, issue 12 Alert me to new issues of AAC About AAC Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy AAC RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0066-4804 Online ISSN: 1098-6596 Copyright © 2011 by the American Society for Microbiology. For an alternate route to AAC .asm.org, visit: http://intl- AAC .asm.org | More Info» var gaJsHost = (("https:" == document.location.protocol) ? "https://ssl." : "http://www."); document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); var pageTracker = _gat._getTracker("UA-5821458-3"); pageTracker._trackPageview(); http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Antimicrobial Agents and Chemotherapy American Society For Microbiology

Purification and characterization of inducible beta-lactamases in Aeromonas spp.

Abstract

Purification and characterization of inducible beta-lactamases in Aeromonas spp. J P Iaconis and C C Sanders Department of Medical Microbiology, Creighton University School of Medicine, Omaha, Nebraska 68178. ABSTRACT beta-Lactamases from Aeromonas hydrophila and A. sobria were purified and characterized. Both species produced beta-lactamases that were inducible by either cefoxitin or imipenem. These species were resistant to ampicillin and cephalothin but not imipenem. Isoelectric focusing of sonic extracts revealed one band at pI 8.0 and a second band at pI 7.0 for A. hydrophila. Likewise, A. sobria produced two bands, one at pI 8.4 and the other at pI 7.0. Two enzymes from each species were separated by flatbed electrofocusing gel and purified to homogeneity. The molecular weight of the pI 7.0 enzyme (A1) from both species was estimated to be 42,500, whereas the pI 8.0 (A2h) and 8.4 (A2s) enzymes of A. hydrophila and A. sobria had molecular weights of 31,500 and 35,000, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The relative Vmax values for cephalothin, penicillin, and imipenem for these enzymes indicated that A1 was primarily a cephalosporinase while A2h and A2s were penicillinases highly active against carbapenems. A1 was susceptible to inhibition by cloxacillin, while the A2 enzymes were inhibited by clavulanic acid and EDTA and required zinc for activity. Thus, there appear to be two distinct inducible beta-lactamases in A. hydrophila and A. sobria that play an important role in the beta-lactam resistance of these species. CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? « Previous | Next Article » Table of Contents This Article doi: 10.1128/​AAC.34.1.44 Antimicrob. Agents Chemother. January 1990 vol. 34 no. 1 44-51 » Abstract PDF Classifications Research Article Services Email this article to a colleague Similar articles in ASM journals Alert me when this article is cited Alert me if a correction is posted Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of AAC Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Load citing article information Citing articles via Web of Science Citing articles via Google Scholar Google Scholar Articles by Iaconis, J. P. Articles by Sanders, C. C. Search for related content PubMed PubMed citation Articles by Iaconis, J. P. Articles by Sanders, C. C. Related Content Load related web page information Social Bookmarking CiteULike Connotea Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter What's this? current issue December 2011, volume 55, issue 12 Alert me to new issues of AAC About AAC Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy AAC RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0066-4804 Online ISSN: 1098-6596 Copyright © 2011 by the American Society for Microbiology. For an alternate route to AAC .asm.org, visit: http://intl- AAC .asm.org | More Info» var gaJsHost = (("https:" == document.location.protocol) ? "https://ssl." : "http://www."); document.write(unescape("%3Cscript src='" + gaJsHost + "google-analytics.com/ga.js' type='text/javascript'%3E%3C/script%3E")); var pageTracker = _gat._getTracker("UA-5821458-3"); pageTracker._trackPageview();
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