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

Abstract:

:A protein-engineered beta-lactamase, constructed by site-directed mutagenesis in Escherichia 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 beta-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.

authors

García JL,Nuñez CJ,González EG,Osuna J,Soberón X,Galindo E

doi

10.1007/BF02785659

subject

Has Abstract

pub_date

1998-05-01 00:00:00

pages

243-56

issue

2-3

eissn

0273-2289

issn

1559-0291

journal_volume

73

pub_type

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