• Title/Summary/Keyword: Acinetobactor sp. B2

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Purification and Characterization of the Lipase from Acinetobacter sp. B2

  • Sohn, Sung-Hwa;Park, Kyeong-Ryang
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.189-195
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    • 2005
  • Industrial development has increase consumption of crude oil and environmental pollution. A large number of microbial lipolytic enzymes have been identified and characterized to date. To development for a new lipase with catalytic activity in degradation of crude oil as a microbial enzyme, Acinetobactor sp. B2 was isolated from soil samples that were contaminated with oil in Daejon area. Acinetobactor sp. B2 showed high resistance up to 10 mg/mL unit to heavy metals such as Ba, Li, Al, Cr, Pb and Mn. Optimal growth condition of Acinetobactor sp. B2 was confirmed $30^{\circ}C$. Lipase was purified from the supernatant by Acinetobactor sp. B2. Its molecular mass was determined to the 60 kDa and the optimal activity was shown at $40^{\circ}C$ and pH 10. The activation energies for the hydrolysis of p-nitrophenyl palmitate were determined to be 2.7 kcal/mol in the temperature range 4 to $37^{\circ}C$. The enzyme was unstable at temperatures higher than $60^{\circ}C$. The Michaelis constant $(K_{m})\;and\;V_{max}$ for p-nitrophenyl palmitate were $21.8{\mu}M\;and\;270.3{\mu}M\;min^{-1}mg\;of\;protein^{-1}$, respectively. The enzyme was strongly inhibited by $Cd{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},\;EDTA$, 2-Mercaptoethalol. From these results, we suggested that lipase purified from Acinetobactor sp. B2 should be able to be used as a new enzyme for degradation of crude oil, one of the environmental contaminants.

Purification and Characterization of Lipase from Acinetobacter sp. B2 Isolated from Oil­contaminated Soil (유류오염지역에서 분리한 Acinetobacter sp. B2로부터의 Lipase 정제 및 특성)

  • Son Seung Hwa;Park Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.320-327
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    • 2004
  • Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil sam­ples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of p­nitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.

Isolation and Identification of the Origins Causing the Slime Found in Pulp and Paper Making Processes (제재공장내 슬라임 발생원의 분리와 동정)

  • Oh, Jung-Soo;Jo, Byoung-Muk;Kim, Eun-Hee
    • Journal of the Korean Wood Science and Technology
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    • v.25 no.3
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    • pp.50-57
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    • 1997
  • The presence of slime in paper mills is practically universal. Many researches have been performed for many years to resolve the problem caused by the slime in pulp and paper mill. Many papers have been published to show the bacteria is a major cause of paper mill slime. Now that the recycling of the water has been increased and the regulations of a toxic chemical dosage have become more strengthen, the importance of the control of slime in pulp and paper mill recently has been more recognized. Therefore, to produce quality products at the lowest economic and environmental costs, a through study of the microbial ecology and the indentification of troublesome slime-forming bacteria is a quite necessary. The purpose of this paper is to indentify slime~forming bacteria isolated from the papermaking process. The samples were taken from four parts of making fine paper : machine chest, head box, wire part, white water tank. Machine chest showed the most numbers of bacteria, numbering $2.55{\times}10^7$. The different colony types were taken from the 105 dilution plate. Nine bacteria were identified u sing the Biolog system and the vitek system: 6 gram-negative bacteria, 3 gram-positive bacteria. They are Pseudomonas paucimobilis B., Staphylococcus sp., Acinetobacter calcoaceticus., Pseudomonas cepacia, Actinobaci1lus capsulatus, Acidovorax sp., Flavobacterium sp., and Staphylococcus auricularis in addition to one unidentified sp., Among them. Pseudomonas paucimobillis was found in all places where the samples were taken. And, each parts had the different predominant bacteria in it : Pseudomonas paucimobilis B. in machine chest, Acinetobactor calcoaceticus. in Wire Part and Staphylococcus sp. in head box.

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