Purification and characterization of an alkaline lipase from Pseudomonas aeruginosa isolated from putrid mineral cutting oil as component of metalworking fluid

Karadžić, Ivanka; Masui, Akihiko; Izrael-Živković, Lidija; Fujiwara, Nobuaki

doi:10.1263/jbb.102.82

Cited by 118 publications

(79 citation statements)

References 26 publications

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“…Lipase activity was measured by the alkaline titration method (19). A solution was prepared by mixing 10 ml of 5% (v/v) substrate (with 1% gum arabic) 10 ml of 50 mM Tris-HCL buffer supplemented with 40 mM sodium chloride (pH 9.0), 2.0 ml of sodium deoxycholate solution (80 mg/ml pH 9.0) and 1 ml of purified enzyme in Tris-HCL buffer (pH 6.8).…”

Section: Substrate Specificitymentioning

confidence: 99%

Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT 9

Borkar¹,

Bodade²,

Rao³

et al. 2009

Braz. J. Microbiol.

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An extra cellular lipase was isolated and purified from the culture broth of Pseudomonas aeruginosa SRT 9 to apparent homogeneity using ammonium sulfate precipitation followed by chromatographic techniques on phenyl Sepharose CL-4B and Mono Q HR 5/5 column, resulting in a purification factor of 98 fold with specific activity of 12307.8 U/mg. The molecular weight of the purified lipase was estimated by SDS-PAGE to be 29 kDa with isoelectric point of 4.5. Maximum lipase activity was observed in a wide range of temperature and pH values with optimum temperature of 55ºC and pH 6.9. The lipase preferably acted on triacylglycerols of long chain (C14-C16) fatty acids. The lipase was inhibited strongly by EDTA suggesting the enzyme might be metalloprotein. SDS and metal ions such as Hg , Ag 2+ and Fe 2+ decreased the lipase activity remarkedly. Its marked stability and activity in organic solvents suggest that this lipase is highly suitable as a biotechnological tool with a variety of applications including organo synthetic reactions and preparation of enantiomerically pure pharmaceuticals. The Km and Vmax value of the purified enzyme for triolein hydrolysis were calculated to be 1.11 mmol/L and 0.05 mmol/L/min respectively.

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Section: Substrate Specificitymentioning

confidence: 99%

Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT 9

Borkar¹,

Bodade²,

Rao³

et al. 2009

Braz. J. Microbiol.

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“…At pH 8, it lost up to 50% activity at the end of a 12 h exposure (Dalal et al, 2008). In another study, Karadzic et al (2006) isolated and purified an extracellular lipase from Pseudomonas aeruginosa. The optimum pH and temperature were 11 and 70 C, respectively.…”

Section: Discussionmentioning

confidence: 99%

Cloning and characterization of two functionally diverse lipases from soil metagenome

Khan

Kottur

Mookambikay

2013

J. Gen. Appl. Microbiol.

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IntroductionLipases are ubiquitous enzymes that catalyze the hydrolysis of fats and oils and form a large group of the enzymes. These enzymes also catalyze the hydrolysis of a wide range of carboxyl esters at the water lipid interface and reverse the reaction in the nonaqueous phase. During the last decade, lipases have gained importance, to a certain extent, over proteases and amylases, especially in food industry. Many lipases have been isolated from bacteria, fungi, plants, and animals, and have received much attention as biocatalysts. The enantioselective and regioselective nature of lipases has been used for the resolution of chiral drugs, fat modification, synthesis of cocoa butter substituent, biofuels, and flavor enhancers. Owing to their novel and multifunctional application, lipase-catalyzed processes have received great attention, mainly due to several advantages: (1) they are more environmentfriendly than bulk chemical syntheses, (2) they allow manufacture of higher quality products, (3) they are easy to recover and re-use and (4) they can be used in continuous operations.Although the microbial diversity in nature has been estimated by culture-independent methods, it is uncertain whether it would be possible to find a universal J. Gen. Appl. Microbiol., 59, 21 31 (2013) A soil metagenomic library was constructed and two functionally diverse lipase genes, SMlipB and SMlipD, were screened by a function-driven approach and characterized. The optimal temperature for enzyme activity of SMlipB and SMlipD was 50°C and 30°C, respectively, and optimal pH was determined to be 7.0 and 9.0, respectively. Both enzymes exhibited broad substrate specificity and showed enhanced activity in the presence of SDS and Tween 20. The SMlipB enzyme was highly resistant to many organic solvents, especially isopropanol, ethanediol, DMSO, methanol and xylene, whereas SMlipD activity was inhibited in all the solvents except xylene.

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“…Lipase activity was completely inhibited in the presence of 1% SDS. It is possible that, besides preventing aggregation of the lipase, the nonionic detergents, such as surface-active substances, stabilize the interfacial area, facilitating the access of the substrate to the enzyme, since the catalytic reaction of lipases takes place at an oil-water interface (Karadzic et al, 2006) …”

Section: Effects Of Inhibitorsmentioning

confidence: 99%

Differences in hydrolytic abilities of two crude lipases from Geotrichum candidum 4013

Brabcová

Zarevúcká

Macková

2010

Yeast

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The fungus Geotrichum candidum 4013 produces two types of lipases (extracellular and cell-bound). Both enzymes were tested for their hydrolytic ability to pnitrophenyl esters and compounds having a structure similar to the original substrate (triacylglycerols). Higher lipolytic activity of extracellular lipase was observed when triacylglycerols of medium-(C12) and long-(C18) chain fatty acids were used as substrates. Cell-bound lipase preferentially hydrolysed trimyristate (C14). The differences in the abilities of these two enzymes to hydrolyse p-nitrophenyl esters were observed as well. The order of extracellular lipase hydrolysis relation velocity was as follows: p-nitrophenyl decanoate > p-nitrophenyl caprylate > p-nitrophenyl laurate > p-nitrophenyl palmitate > p-nitrophenyl stearate. The cell-bound lipase indicates preference for p-nitrophenyl palmitate. The most striking differences in the ratios between the activity of both lipases (extracellular : cell-bound) towards different fatty acid methyl esters were 2.2 towards methyl hexanoate and 0.46 towards methyl stearate (C18). The Michaelis constant (K m ) and maximum reaction rate (V max ) for p-nitrophenyl palmitate hydrolysis of cell-bound lipase were significantly higher (K m 2.462 mM and V max 0.210 U/g/min) than those of extracellular lipase (K m 0.406 mM and V max 0.006 U/g/min).

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Purification and characterization of an alkaline lipase from Pseudomonas aeruginosa isolated from putrid mineral cutting oil as component of metalworking fluid

Cited by 118 publications

References 26 publications

Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT 9

Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT 9

Cloning and characterization of two functionally diverse lipases from soil metagenome

Differences in hydrolytic abilities of two crude lipases from Geotrichum candidum 4013

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