Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Effect of plasmid curing on the 2, 3-dihydroxybenzoic acid production and antibiotic resistance of Acinetobacter sp. B-W

Acinetobacter sp. B-W의 2, 3-dihydroxybenzoic acid 생산과 항생제 저항성에 미치는 플라스미드 제거 효과

  • Kim, Kyoung-Ja (Department of Life Science and Biotechnology, College of Natural Science, Soonchunhyang University) ;
  • Kim, Jin-Woo (Department of Life Science and Biotechnology, College of Natural Science, Soonchunhyang University) ;
  • Yang, Yong-Joon (Department of Plant and Food Science, Sangmyung University)
  • 김경자 (순천향대학교 생명시스템학과) ;
  • 김진우 (순천향대학교 생명시스템학과) ;
  • 양용준 (상명대학교 식물식품공학과)
  • Received : 2016.07.27
  • Accepted : 2016.08.08
  • Published : 2016.09.30
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Abstract

Acinetobacter sp. B-W producing siderophore, 2, 3-dihydroxybenzoic acid (DHB) was analyzed for plasmid content. Strain B-W harbored plasmid of 20 kb in size. Growth at $43^{\circ}C$ was effective in producing mutant cured of plasmid of strain B-W. This mutant lost the ability to produce 2, 3-DHB. Formation of siderophore halos on the chrome azurol S (CAS) agar medium was not detected by cured strain B-W. pHs of supernatants of wild type strain B-W and cured mutant grown in glucose and $MnSO_4$ containing medium at $28^{\circ}C$ for 3 days were 4.5 and 8.5, respectively. Antibiotic resistance against ampicillin, actinomycin D, bacitracin, lincomycin, and vancomycin was lost in cured mutant. Plasmid curing of strain B-W resulted in drastic reduction of minimal inhibitory concentration (MIC) of several antibiotics. E. coli $DH5{\alpha}$ was transformed with plasmid isolated from strain B-W. The transformant E. coli $DH5{\alpha}$ harbored a plasmid of the same molecular size as that of the donor plasmid. Transformant E. coli $DH5{\alpha}$ produced 2, 3-DHB and contained antibiotic resistant ability. Thus a single plasmid of 20 kb seemed to be involved in 2, 3-DHB production. Genes encoding resistance to antibiotics were also supposed to be located on this plasmid.

시드로포어인 2, 3-dihydroxybenzoic acid (DHB)를 생산하는 Acinetobacter sp. B-W의 플라스미드를 분석한 결과, 20 kb 크기의 플라스미드를 함유하였다. 배양 온도 $43^{\circ}C$ 가 플라스미드가 제거된 돌연변이체 생산에 효과적이었다. 이 돌연변이체는 2,3-DHB 생산 능력을 소실하였으며, chrome azurol S (CAS) 아가 배지에서 시드로포어 생산이 검출되지 않았다. 포도당과 황산 망간을 함유한 배지에서 $28^{\circ}C$로 3일간 배양한 B-W 원 균주와 돌연변이체의 배양 상등액의 pH는 각각 4.5와 8.5로 나타났다. 돌연변이체에서는 ampicillin, actinomycin D, bacitracin, lincomycin과 vancomycin 같은 항생제에 대한 저항성이 사라졌으며, 이러한 항생제에 대한 최소 억제 농도(MIC)가 급격하게 감소하였다. B-W 균주에서 분리한 플라스미드로 대장균을 형질전환시킨 결과, 원 균주와 같은 크기의 플라스미드가 이 형질전환 대장균에서 발견되었다. 플라스미드가 제거된 돌연변이체에서는 플라스미드가 발견되지 않았다. 20 kb 크기의 플라스미드에 2,3-DHB 생산 유전자와 여러 항생제 저항성 유전자가 자리잡고 있는 것으로 추정된다.

Keywords

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