Calorimetric analysis of cephalosporins using an immobilized TEM-1 beta-lactamase on Ni2+ chelating sepharose fast flow

Research output: Contribution to journalArticle

Abstract

Two beta-lactamases, penicillinase type I from Bacillus cereus and TEM-1 beta-lactamase from Haemophilus ducreyi, were immobilized on a Chelating Sepharose Fast Flow column loaded with Ni2+ in an active form. Flow-injection analysis of beta-lactams was performed by using an enzyme column reactor fitted into the enzyme thermistor. With both enzymes it was possible to monitor both penicillins and cephalosporins. Moreover, Michaelis constants of the TEM-1 beta-lactamase were markedly increased upon immobilization for all substrates, especially carbenicillin, cephaloridine, and cefoperazone.

Details

Authors
  • R Lawung
  • Bengt Danielsson
  • V Prachayasittikul
  • L Bülow
Organisations
External organisations
  • Mahidol University
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Analytical Chemistry
  • Biochemistry and Molecular Biology

Keywords

  • Anti-Bacterial Agents, Bacillus cereus, Calorimetry, Carbenicillin, Cefoperazone, Cephaloridine, Cephalosporins, Chelating Agents, Chromatography, Affinity, Chromatography, Agarose, Enzymes, Immobilized, Haemophilus ducreyi, Nickel, Penicillinase, Penicillins, beta-Lactamases, beta-Lactams
Original languageEnglish
Pages (from-to)57-62
Number of pages6
JournalAnalytical Biochemistry
Volume296
Issue number1
StatePublished - 2001 Sep 1
Publication categoryResearch
Peer-reviewedYes