• Journal of Life Science
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• v.23 no.10
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• pp.1246-1251
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• 2013

An organic solvent-tolerant lipase of Pseudomonas sp. BCNU 154 that was isolated from wastewater in the industrial complex region had optimal activity at $37^{\circ}C$ and pH 8. This crude extracellular lipase from BCNU 154 exhibited maximum stability in toluene, retaining about 6.01 U/ml (117.53%) activity for 2 h. $Ca^{2+}$, $Mg^{2+}$, $NH_4{^+}$, and $Na^+$ ions and triton X-100 activated the enzymes, whereas $Ba^{2+}$, $Hg^{2+}$, and $Zn^{2+}$ ions inhibited their activity. Pseudomonas sp. BCNU 154 lipase revealed stable activity comparable to that of the commercial immobilized Novozym 435. Thus, this organic solvent-tolerant lipase could have potential as a whole cell biocatalyst in industrial chemical processes without the use of immobilization.

• Microbiology and Biotechnology Letters
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• v.21 no.3
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• pp.239-246
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• 1993

Producing an extracellular alkaline lipase, this isolate JM123 was identified as a Pseudomonas aeruginosa strain from the results of the analyses of its morphological, biochemical and physiological properties. This strain showed the highest productivity of alkaline lipase when grown at pH 9.0 and 30C for 13-20 hours in the medium of 2% starch, 1% soytone, 0.5% peptone and 1% MgSO4.7H2O. However, this enzyme was greatly repressed when grown in the glucose containing medium. The culture broth was fractionated by the order of the ammonium sulfate precipitation, Sephadex G-200 gel filtration, DEAE-cellulose column chromatography, and Sephadex G-150 gel filtration.

• Journal of Life Science
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• v.26 no.5
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• pp.603-607
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• 2016

A crude extracellular lipase from solvent-tolerant bacterium Pseudomonas sp. BCNU 106 was highly stable in the broad pH range of 4-10 and at temperature of 37℃. Crude lipase of BCNU 106 exhibited enhanced stability in 25% organic solvents such as xylene (121.85%), hexane (120.35%), octane (120.41 %), toluene (118.14%), chloroform (103.66%) and dodecane (102.94%) and showed excellent stability comparable with the commercial immobilized enzyme. In addition, the stability of BCNU 106 lipase retained above 110% of its enzyme activity in the presence of Cu²⁺, Hg²⁺, Zn²⁺ and Mn²⁺, whereas Fe²⁺ strongly inhibited its stability. The detergents including tween 80, triton X-100 and SDS were positive signals for lipase stability. Because of its stability in multiple organic solvents, cations and surfactants, the Pseudomonas sp. BCNU 106 lipase could be considered as a potential biocatalyst in the industrial chemical processes without using immobilization.

• Journal of Life Science
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• v.22 no.3
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• pp.306-311
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• 2012

A gene encoding a lipase, lipT, was cloned from the psychrotrophic bacterium Pseudomonas mandelii and sequenced. An open reading frame of 1,686 bp was found that encodes a polypeptide consisting of 562 amino acid residues. Sequence analysis revealed a Gly-His-Ser-Leu-Gly sequence, which matches the consensus Gly-X-Ser-X-Gly motif conserved among lipolytic enzymes. The recombinant LipT protein was predominantly expressed as inclusion bodies in Escherichia coli and subsequently purified by nickel-chelate affinity chromatography. A small fraction of LipT was refolded, and the subsequent LipT exhibited substrate preferences for p-nitrophenyl butyrate (C4) and p-nitrophenyl octanoate (C8).

• Microbiology and Biotechnology Letters
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• v.22 no.3
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• pp.271-276
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• 1994

The strain S4-14 which produced alkaline lipase and had resistance against linear alkylbenzene sulfonate was isolated from soil or water samples. The isolated strain S4-14 was identified a species belong to Pseudomonas. Alkalin lipase secreted by Pseudomonas sp. S4-14 was purified by ammonium sulfate precipitation procedure follwed by DEAE-Cellulose, DEAE-Sepharose and gel filtration chromatohraphies with 995.15 U/mg protein and 16.1% yield. The molecular weight of the enzyme was estimated to be 65,000 dalton by SDS-PAGE. The optimum pH and temperature of the purified enzyme was 10.5 and 45$\circ $C, respectively. The emzyme was stable at 45$\circ $C for 1 hr and in a pH range from 8.0 to 12.0 for 24 hr at 4$\circ $C. The activity of lipase was enhanced by Ca$^{2+}$ while inhibited strongly by Pb$^{2+}$, Zn$^{2+}$ or Fe$^{3+}$. The activity of lipase was inactivated about 50~60% in the presence of 50 mg/l linear alkylbenzene sulfonate, $\alpha $-olefin sulfonate, alcohol ethoxylate or perborate.

• Bulletin of the Korean Chemical Society
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• v.24 no.9
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• pp.1269-1275
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• 2003

The quaternary carbon-containing alcohols (1-6) were resolved enantioselectively by various lipases such as PFL (Pseudomonas fluorescens lipase), LAK (Pseudomonas fluorescens lipase), CRL (Candida rugosa lipase) and PCL (Pseudomonas cepacia lipase). The enzymatic resolution of racemic alcohol $({\pm})-2$ gave the excellent enantioselectivity in favor of (S)-2d in 99% ee, while those of the racemic alcohols (1, 3, 4, 5 and 6) gave the resolved alcohols with moderate to good enantioselectivity. Also, their absolute configurations were determined by chemical transformation to the known compounds.

• Journal of Life Science
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• v.25 no.3
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• pp.345-348
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• 2015

An organic solvent stable lipase from solvent-tolerant Pseudomonas sp. BCNU 171 had an optimal pH of 8 and an optimal temperature of 37℃. This crude extracellular lipase from BCNU 171 exhibited increased stability in the presence of various types of solvents at high concentrations (25%, v/v). The lipase stability was found to be highest in the presence of xylene (137%), followed by toluene (131%), octane (130%), and butanol (104%). Overall, BCNU 171 lipase tended to be more stable than immobilized commercial lipase (Novozyme435) in the presence of organic solvents. Furthermore, BCNU 171 lipase maintained about 90% of its enzyme original activity in the presence of NH₄⁺, Na⁺, Ba²⁺, Hg²⁺, Ni²⁺, Cu²⁺, and Ca²⁺ion and significantly increased its enzyme activity in the presence of various emulsifying agents. Thus, the organic solvent stable lipase from Pseudomonas sp. BCNU 171 could be usable as a potential whole cell biocatalyst and for synthetic applications of enzymes for industrial chemical processes in organic solvents without using immobilization.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.57-63
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• 1991

A strain J-19 was isolated from soil, produced lipase which has resistant against alkali and linear alkylbenzene sulfonate. The strain was identified as Pseudornonns sp.. The enzyme was purified by ammonium sulfate precipitation, DEAE-Sephadex and Sephadex G- 100 column chromatography. The specific activity of the purified enzyme was 35 unit/mg protein and the yield of enzyme activity was 17%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis. Mo1ecul;tr weight of the purified enzyme was estimated about 36,000 by Sephadex GI00 gel filtration and SDS-polyacrylarnide gel electrophoresis. The optimum pH and temperature were pH 10.0 and $30^{\circ}C$, respectively. Activity of the purified enzyme was increased 2-fold by the addition of 0.1% linear alkylbenzene sulfonate and 2.5- fold by the addition of 0.05% Tide. This enzyme remained stable from pH 8.0 to 10.0 and stable up to $40^{\circ}C$.

• Journal of Microbiology
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• v.41 no.2
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• pp.95-101
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• 2003

The lipase gene (lipA) and its activator gene (lipB) of Pseudomonas sp. SW-3 were cloned and sequenced. The lipB was found to be present immediately downstream of lipA. The deduced amino acid sequences of lipA and lipB showed a high level of homology to those of other lipases belonging to the family I.1 of bacterial lipases. When lipA was expressed in Escherichia coli using T7 promoter, an active lipase was produced in cells carrying both lipA and lipB, but not in cells harboring only lipA. Recombinant lipase (rPSL) overproduced in an insoluble form was solubilized in the presence of 8 M urea, purified in a urea-denatured form and refolded by removing urea in the presence of the Ca$\^$2+/ ion. rPLS had maximum activity at pH 8.0 and 50$^{\circ}C$, was stable at pHs from 7.0 to 9.0 and below 50$^{\circ}C$, and showed the highest activity toward the p-nitrophenyl ester of palmitate (Cl6).

• Journal of Microbiology
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• v.44 no.6
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• pp.583-590
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• 2006

In this study, an organic solvent tolerant bacterial strain was isolated. This strain was identified as Pseudomonas sp. strain S5, and was shown to degrade BTEX (Benzene, Toluene, Ethyl-Benzene, and Xylene). Strain S5 generates an organic solvent-tolerant lipase in the late logarithmic phase of growth. Maximum lipase production was exhibited when peptone was utilized as the sole nitrogen source. Addition of any of the selected carbon sources to the medium resulted in a significant reduction of enzyme production. Lower lipase generation was noted when an inorganic nitrogen source was used as the sole nitrogen source. This bacterium hydrolyzed all tested triglycerides and the highest levels of pro-duction were observed when olive oil was used as a natural triglyceride. Basal medium containing Tween 60 enhanced lipase production to the most significant degree. The absence of magnesium ions ($Mg^{2+}$) in the basal medium was also shown to stimulate lipase production. Meanwhile, an alkaline earth metal ion, $Na^+$, was found to stimulate the production of S5 lipase.