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

다국어 초록 (Multilingual 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.

참고문헌 (Reference)

1 Schwyn, R., "Universal chemical assay for detection and determination of siderophores" 160 : 47-56, 1987

2 김경자, "Temperature dependent 2,3-dihydroxybenzoic acid production in Acinetobacter sp. B-W" 한국미생물학회 51 (51): 249-255, 2015

3 Jalal, M. A. F., "Structure of anguibactin, a uique plasmid related bacterial siderophore from the fish pathogen Vibrio anguillarum" 111 : 292-296, 1989

4 Kenchappa, P., "Simplified panel of assimilation tests for identification of Acinetobacter species" 246 : 700-706, 2003

5 Meyers, J. A., "Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid" 123 : 1529-1537, 1976

6 Miethke, M., "Siderophore-based iron acquisition and pathogen control" 71 : 413-451, 2007

7 Driss, F., "Relationship between plasmid loss and gene expression in Bacillus thuringiensis" 62 : 1287-1293, 2011

8 Young, I.G., "Regulation of the enzymes involved in the biosynthesis of 2, 3-dihydroxybenzoic acid in Aerobacter aerogenes and E. coli" 177 : 401-411, 1969

9 Clinical and Laboratory Standards Institute, "Performance Standards for Antimicrobial Susceptibility Testing: Twentieth Informational Supplement M100-S20"

10 Saranathan, R., "Multiple drug resistant carbapenemases producing Acinetobacter baumannii isolates harbours multiple R-plasmids" 140 : 262-270, 2014

11 Maniatis, T., "Molecular cloning:A Laboratory Manual" Cold Spring Harbor Laboratory Press 1989

12 Wencewicz, T., "Is drug release necessary for antimicrobial activity of siderophore-drug conjugates? Syntheses and biological studies of the naturally occurring salmycin “Trojan Horse” antibiotics and synthetic desferridanoxamine antibiotic conjugates" 22 : 633-648, 2009

13 Zhang, R., "High incidence of plasmids in marine Vibrio species isolated from Mai Po Nature Reserve of Hong Kong" 21 : 1661-1668, 2012

14 Zurkowski, W., "Effective method for the isolation of non-nodulation mutants of Rhizobium trifolii" 32 : 311-314, 1978

15 Elias, H. M., "Determination of plasmid DNA role in multidrug resistant Pseudomonas aeruginosa clinical isolates" 1 : 80-86, 2013

16 González, J. M., "Correlation between specific plasmids and delta-endotoxin production in Bacillus thuringiensis" 5 : 351-365, 1981

17 Arnow, L. E, "Colorimetric determination of the components of 3, 4-dihydroxyphenylalanine-tyrosine mixtures" 18 : 531-537, 1937

18 Chen, Q., "Chromosome-mediated 2, 3-dihydroxybenzoic acid is a precursor in the biosynthesis of the plasmid mediated siderophore anguibactin in Vibrio anguillarum" 176 : 4226-4234, 1994

19 Wandersman, C., "Bacterial iron sources: from siderophores to hemophores" 58 : 611-647, 2004

20 Bauer, A. W., "Antibiotic susceptibility testing by a standardized single disk method" 45 : 493-496, 1966

21 Steers, E., "An inocula replicating apparatus for routine testing of bacterial susceptibility to antibiotics" 9 : 307-311, 1959