Calorimetric Analysis of Cephalosporins Using an Immobilized TEM-1 β-Lactamase on Ni2+ Chelating Sepharose Fast Flow
✍ Scribed by Ratana Lawung; Bengt Danielsson; Virapong Prachayasittikul; Leif Bülow
- Book ID
- 102560176
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 81 KB
- Volume
- 296
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Two -lactamases, penicillinase type I from Bacillus cereus and TEM-1 -lactamase from Haemophilus ducreyi, were immobilized on a Chelating Sepharose Fast Flow column loaded with Ni 2؉ in an active form. Flow-injection analysis of -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 -lactamase were markedly increased upon immobilization for all substrates, especially carbenicillin, cephaloridine, and cefoperazone.