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KMID : 0043319980210030310
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Archives of Pharmacal Research
1998 Volume.21 No. 3 p.310 ~ p.314
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Resistance Mechanism of Acinetobacter spp. Strains Resistant to DW-116, a New Quinolone
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Choi Keum-Hwa
Baek Moon-Chang
Kim Byung-Kak
Choi Eung-Chil
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Abstract
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DW-116 is a new fluoroquinolone antimicrobial agent with a broad spectrum. In order to elucidate the resistance mechanism to DW-116 in Acinetobacter spp. bacteria, total chromosomal DNA was isolated from 10 strains of Acinetobacter spp. resistant to DW-116. Quinolone resistance determinant region (QRDR) of DNA gyrase gene was amplified by PCR. The 345 bp nucleotide fragment yielded was inserted into pKF 3 which was used as the vector. Comparisons of the DNA sequences of 8 strains with that of the wild type strain revealed a Ser-83 to Leu mutation in mutants and all ten strains contained one silent mutationin QRDR. From Acinetobacter MB4-8 strain, DNA gyrase was isolated and purified, through novobiocin-sepharose, heparin-sepharose affinity column chromatography. The enzyme was composed of two subunits and the molecular mass of subunits A and B were 75.6 and 51.9 kDa, respectively. The supercoiling activity of the reconstituted DNA gyrase composed of subunit A from Acinetobacter MB4-8 and subunit B from E. coli was not inhibited by ml of ciprofloxacin. It might be said that one of the resistance mechanisms to DW-116 in Acinetohacter MB4-8 was subunit A alteration of DNA gyrase.
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KEYWORD
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DW-116, Fluoroquinolne, PCR, DNA gyrase, QRDR, Acinetobacter, Ciprofloxacin
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