• Journal of Life Science
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• v.29 no.4
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• pp.492-498
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• 2019

Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.

• Korean Journal of Microbiology
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• v.43 no.1
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• pp.66-71
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• 2007

Two isolates of genus Acinetobacter were obtained from Jang-Baek waterfall in North Korea. Morphological characteristics of the isolated 2 strains were Gram-negative, aerobic and rod shape bacteria. Physiological and biochemical characterization of the isolated 2 strains were some different aspect from those of type strains. 16S rDNA sequence analysis showed that the two isolates shared 99.9% sequence similarity. Strains JB10 and $JB15^{T}$ were shown to belong to the Gammaproteobacteria and showed the highest levels of sequence similarity to Acinetobacter tandoii $4N13^{T}$ (97.3%), Acinetobacter haemolyticus $ATCC17906^{T}$ (97.2%), Acinetobacter johnsonii $DSM6963^{T}$ (97.2%), Acinetobacter junii $DSM6964^{T}$ (96.7%), Acinetobacter schindleri $LUH5832^{T}$ (97.0%) and Acinetobacter ursingii $LUH3792^{T}$ (96.6%). The major cellular fatty acid in Acinetobacter type strains and isolated strains included $C_{18:1}\;{\omega}9c\;and\;C_{16:1}\;{\omega}7c/C_{15:0}\;iso\;2OH$. Eventhough it was ascertained that the isolated strains were closely related to genus Acinetobacter, physiological and biochemical characteristics and the result of the isolated strains 16S rDNA analysis indicate some different aspects from those of type strains of genus Acinetohacter It is considered that the isolated JB10 (=KEMC 52-093) and $JB15^{T}\;(=KEMC\;52-094^{T})$ strains be new species of genus Acinetobacter. We name it as Acinetobacter koreensis sp. nov.

• Journal of Veterinary Clinics
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• v.31 no.1
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• pp.36-39
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• 2014

Many bacteria colonized in the horse semen affect quality of the sperm and some may cause infection in the mare reproductive tract and infertility of susceptible mare. This study was initiated to determine the prevalence of bacteria in testes of Jeju horses by determining rRNA sequence. The samples were swabed from the testes of nine Jeju horses (aged from 8 to 12 months after birth). Bacteria isolated from testes were identified by 16S rDNA sequencing. 1.6-kbp PCR products for 16S rRNA coding region were obtained using the universal primers. The PCR products were further purified and sequenced. Maximum similar species were found by BLAST search in the GenBank DNA database. BLAST results showed that the sequences were similar to those of Acinetobacter sp (A. schindleri, A. ursingii)., Bacillus cereus, Corynebacterium glutamicum, Escherichia coli, Gamma proteobacterium, Micrococcus luteus, Pseudomonas mendocina, Shigella sonnei, Sphingomonas sp., Staphylococcus sp (S. cohnii, S. saprophyticus, S. xylosus)., and Stenotrophomonas maltophilia. DNA sequences for 16S rRNA is provided useful informations for species identification of pathogenic microorganisms for the reproductive organs in horses.