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Antimicrobial Agents and Chemotherapy, February 1999, p. 307-313, Vol. 43, No. 2
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Characterization of SFO-1, a Plasmid-Mediated Inducible Class A beta -Lactamase from Enterobacter cloacae

Yoshimi Matsumoto1,2,* and Matsuhisa Inoue1

Department of Microbiology, Kitasato University School of Medicine, Sagamihara,1 and Medicinal Biology Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., Osaka,2 Japan

Received 1 April 1998/Returned for modification 23 July 1998/Accepted 24 November 1998

Enterobacter cloacae 8009 produced an inducible class A beta -lactamase which hydrolyzed cefotaxime efficiently. It also hydrolyzed other beta -lactams except cephamycins and carbapenems. The activity was inhibited by clavulanic acid and imipenem. The bla gene was transferable to Escherichia coli by electroporation of plasmid DNA. The molecular mass of the beta -lactamase was 29 kDa and its pI was 7.3. All of these phenotypic characteristics of the enzyme except for inducible production resemble those of some extended-spectrum class A beta -lactamases like FEC-1. The gene encoding this beta -lactamase was cloned and sequenced. The deduced amino acid sequence of the beta -lactamase was homologous to the AmpA sequences of the Serratia fonticola chromosomal enzyme (96%), MEN-1 (78%), Klebsiella oxytoca chromosomal enzymes (77%), TOHO-1 (75%), and FEC-1 (72%). The conserved sequences of class A beta -lactamases, including the S-X(T)-X(S)-K motif, in the active site were all conserved in this enzyme. On the basis of the high degree of homology to the beta -lactamase of S. fonticola, the enzyme was named SFO-1. The ampR gene was located upstream of the ampA gene, and the AmpR sequence of SFO-1 had homology with the AmpR sequences of the chromosomal beta -lactamases from Citrobacter diversus (80%), Proteus vulgaris (68%), and Pseudomonas aeruginosa (60%). SFO-1 was also inducible in E. coli. However, a transformant harboring plasmid without intact ampR produced a small amount of beta -lactamase constitutively, suggesting that AmpR works as an activator of ampA of SFO-1. This is the first report from Japan describing an inducible plasmid-mediated class A beta -lactamase in gram-negative bacteria.


* Corresponding author. Mailing address: Medicinal Biology Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., 1-6, 2-Chome, Kashima, Yodogawa-ku, Osaka, 532 Japan. Phone: (06) 390-1146. Fax: (06) 304-1192. E-mail: yoshimi_matsumoto{at}po.fujisawa.co.jp.


Antimicrobial Agents and Chemotherapy, February 1999, p. 307-313, Vol. 43, No. 2
0066-4804/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



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