Effect of plasmid curing on the 2, 3-dihydroxybenzoic acid production and antibiotic resistance of Acinetobacter sp. B-W
2016
Kim, K.J., Soonchunhyang University, Asan, Republic of Korea | Kim, J.W., Soonchunhyang University, Asan, Republic of Korea | Yang, Y.J., Sangmyung University, Cheonan, Republic of Korea
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 43Celsius 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 MnSO4 containing medium at 28Celsius 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 DH5α was transformed with plasmid isolated from strain B-W. The transformant E. coli DH5α harbored a plasmid of the same molecular size as that of the donor plasmid. Transformant E. coli DH5α 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.
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