Separation of plasmid-mediated extended spectrum beta-lactamases by fast protein liquid chromatography (FPLC system).

Abstract:

:We have devised a reliable procedure for the separation of three beta-lactamases of isoelectric focusing points (pI), 5.4, 6.5, and 7.9 by Fast Protein Liquid Chromatography (FPLC System). All of these enzymes were transferable and originated from a ceftazidime and cefotaxime resistant Klebsiella pneumoniae isolated in Bombay, India. The complete separation of the enzymes, achievable by this method, allowed each of the different individual beta-lactamases to be characterized biochemically. This analysis revealed that the enzymes of pI 6.5 and pI 7.9 hydrolysed ceftazidime and cefotaxime, and were responsible for the resistance of K. pneumoniae, and its Escherichia coli J53-2 transconjugant to third generation cephalosporins. The enzyme of pI 5.4 was the TEM-1 beta-lactamase. The beta-lactamase of pI 7.9 appears quite different from any previously reported third generation cephalosporin hydrolysing beta-lactamase, and consequently given the preliminary designation DJP-1. This is also the first example of extended spectrum hydrolysing beta-lactamases found in Asia.

journal_name

FEMS Microbiol Lett

authors

Payne DJ,Hood J,Marriott MS,Amyes SG

doi

10.1016/0378-1097(90)90064-w

subject

Has Abstract

pub_date

1990-06-01 00:00:00

pages

195-200

issue

3

eissn

0378-1097

issn

1574-6968

journal_volume

57

pub_type

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