Applied Biochemistry and Biotechnology

, Volume 73, Issue 2–3, pp 243–256 | Cite as

Microbial sensor for new-generation cephalosporins based in a protein-engineered β- lactamase

  • Juan L. García
  • Carlos J. Nuñez
  • Edith G. González
  • Joel Osuna
  • Xavier Soberón
  • Enrique Galindo
Original Articles


A protein-engineered β-lactamase, constructed by site-directed mutagenesis inEscherichia coli (E104M/G238S), and having broadened specificity, was able to degrade cephalosporins of first, second, and third generations. Manipulations of culture conditions allowed an increase in β-lactamase specific activity by up to twofold. The resultant bacteria were used to construct an immersable whole-cell biosensor for the detection of new-generation cephalosporins. Cells were immobilized on agar membranes, which in turn were attached to the surface of a flat pH electrode, thus constituting a biosensor based on the detection of pH changes. The sensor was able to detect second- and third-generation cephalosporins: cefamandole (0.4-4 mM), cefotaxime (0.4-3.5 mM), and cefoperazone (0.3-1.85 mM). Response times were between 3.5 and 11 min, depending on the kind of cephalosporin tested. The biosensor was stable for at least 7 d, time during which up to 100 tests were performed.

Index Entries

β-lactamase mutant cephalosporin sensor 


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Copyright information

© Humana Press Inc 1998

Authors and Affiliations

  • Juan L. García
    • 1
  • Carlos J. Nuñez
    • 1
  • Edith G. González
    • 1
  • Joel Osuna
    • 2
  • Xavier Soberón
    • 2
  • Enrique Galindo
    • 1
  1. 1.Department of Bioengineering, Institute of BiotechnologyNational University of MexicoCuernavacaMéxico
  2. 2.Department of Molecular Recognition and Biostructure, Institute of BiotechnologyNational University of MexicoCuernavacaMéxico

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