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

Lawung, R LU ; Danielsson, Bengt LU ; Prachayasittikul, V and Bülow, L LU (2001) In Analytical Biochemistry 296(1). p.57-62
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.

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author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
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
in
Analytical Biochemistry
volume
296
issue
1
pages
6 pages
publisher
Elsevier
external identifiers
  • pmid:11520032
  • scopus:0035450908
ISSN
0003-2697
DOI
10.1006/abio.2001.5226
language
English
LU publication?
yes
id
3299b0f0-77fc-437a-80ff-cf171500b0f0
date added to LUP
2016-04-18 15:56:22
date last changed
2024-01-04 02:08:52
@article{3299b0f0-77fc-437a-80ff-cf171500b0f0,
  abstract     = {{<p>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.</p>}},
  author       = {{Lawung, R and Danielsson, Bengt and Prachayasittikul, V and Bülow, L}},
  issn         = {{0003-2697}},
  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}},
  language     = {{eng}},
  month        = {{09}},
  number       = {{1}},
  pages        = {{57--62}},
  publisher    = {{Elsevier}},
  series       = {{Analytical Biochemistry}},
  title        = {{Calorimetric analysis of cephalosporins using an immobilized TEM-1 beta-lactamase on Ni2+ chelating sepharose fast flow}},
  url          = {{http://dx.doi.org/10.1006/abio.2001.5226}},
  doi          = {{10.1006/abio.2001.5226}},
  volume       = {{296}},
  year         = {{2001}},
}