Purification and characterization of alkaline Lipase from Pseudomonas aeruginosa strain (original) (raw)
Related papers
2018
The present study aimed to Purification and characterization of lipase produced by the isolate Pseudomonas aeruginosa . The results showed that crude lipase was purified by using the precipitation step of several concentrations ratios of cold acetone. The concentration ratio 4:1 gave the best specific activity 6 U/mg of protein. The subsequent purification step using gel filtration was showed that specific activity reached to 40.5U/mg of protein. The gel electrophoresis (SDS-PAGE) of lipase purity were determined. the result showed there is an appearance of a single protein band predominant in all purification steps, While The molecular weight of purified lipase was 68KDa
Isolation, purification and properties of lipase from Pseudomonas aeruginosa
AFRICAN JOURNAL OF BIOTECHNOLOGY, 2012
Six isolates (Ps1, Ps2, Ps3, Ps4, Ps5 and Ps6) producing lipase were screened from wastewater on a selective medium agar containing Tween 80 or olive oil as the only source of carbon. Isolate Ps 5 showed the highest lipase activity which was later identified as Pseudomonas aeruginosa. The effect of media composition was analysed to maximize the production of lipase. The maximum extracellular lipase present in the broth was purified 4 folds with an overall yield of 19.4% through the purification procedure of ammonium sulphate precipitation and diethyl aminoethyl (DEAE) cellulose chromatography. The purified lipase had the maximal activity within the pH range of 6 to 8, with an optimum pH of 7, and within the temperature range of 20 to 35°C, with an optimum temperature for the hydrolysis of olive oil at 30°C. The enzyme activity of P. aeruginosa lipase was enhanced by Ca 2+ and Mg 2+ but strongly inhibited by heavy metals such as Zn 2+ , Cu 2+ and Mn 2+ .
Isolation, Characterization and Purification of Lipase and Its Gene from Pseudomonas Sp. Ras-4
Ghanis are the ancient rich oil extracting units and their soil is contaminated during extraction process. They use stoned crushing unit which will be tied with bull to rotate and extract oil, during this process oil spills are said to be contaminated. The soil samples were collected from oil ghanas of 3 districts -Gulbarga, Raichur and Bellary were screened for the lipase production. The isolated strain Pseudomonas aeruginosa RAS-4 was identified by 16S rRNA sequence and was screened for the production of lipase by Rhodamine B-Olive oil Agar. This exhibited a clear zone of hydrolysis indicating higher lipase activity. In the present study lipase enzyme was partially purified by ammonium sulphate precipitation. 30% ammonium sulphate showed higher activity of the enzyme than 80% in which the enzyme activity was decreased. Effect of various physicochemical parameters such as pH, temperature, metal ions and inhibitors were studied for the lipase production by RAS-4. The optimized conditions were 35 o C, pH 7.0 and the production was enhanced in the presence of Ca ++ and Na + .
Production, optimization and characterization of the lipase from Pseudomonas aeruginosa
2012
Six isolates of lipase producing ( Ps1, Ps2, Ps3, Ps4, Ps5 and Ps6 ) were secreened from wastewater on a selective medium agar that contained Tween 80 or olive oil as the only carbon source. The isolate showed highest lipase activity was Ps5 which was later identified as Pseudomonas sp. The effect of media composition was analyzed to maximize production of lipase. Pseudomonas aeruginosa had maximal activity at the pH range 6 to 8, with an optimum pH of 7, and the temperature range of 20 – 35°C, with optimum temperature for the hydrolysis of olive oil at 30°C. The lipase activity of the enzyme was enhanced by Ca +2 and Mg +2 but strongly inhibited by heavy metals Zn +2 and Cu +2 and Mn +2 .
Production, optimization and purification of lipase from Pseudomonas aeruginosa
African Journal of Microbiology Research, 2012
Microbial lipases today occupy a place of prominence among biocatalysts owing to their ability to catalyze a wide variety of reactions. Out of 14 bacterial isolates obtained from wastewater sample. The isolates were grown on a selective medium agar that contained tween 80 or olive oil as the only source of carbon for detection of lipase activity 6 exhibited lipase activity. Pseudomonas aeruginosa (Ps 5 ) was the maximum clear zone (22 mm) around the colony and was selected for physicochemical studies. It was found that maximum lipase activity was 37 U.ml -1 in liquid medium. Various physicochemical parameters such as pH, temperature and effect of different medium composition were studied in order to determine the optimum conditions for lipase production. The lipase present in the broth was purified with ammonium sulphate precipitation and DEAE cellulose chromatography to obtain 4 folds pure enzymes. The enzyme activity of the P. aeruginosa lipase was enhanced by Ca +2 and Mg +2 but strongly inhibited by heavy metals Zn +2 , Cu +2 and Mn +2 .
Pakistan Journal of Chemistry
The present work focused on the production of extracellular lipase from Pseudomonas aeruginosa HE05 emplying natural oils of plants as substrat. Nine isolates (HE01-HE09) were isolated from the desiel oil contaminated soil and screened for lipase activity on nutrient agar medium supplemented with 1% olive oil and tween 80. Pseudomonas aeruginosa HE05 was identified as the hyperproducer of lipase. Maximum lipase production (3123U/ml) was recorded when 1% olive oil and 1% tween 80 added in nutrient broth used as a substrate in submerged fermentation. The optimal production was achevied in 96 hrs at 37 o C at 100 rpm. Quantification of lipase units were performed by turbidimetric assay. Supplementation of nutrient broth with natural oils was found as inducer as well as substrate for the production of lipase. Among the natural oils, mustard oil, corn oil, almond oil are better substrate for extracellular lipase production.
Journal of the Taiwan Institute of Chemical Engineers, 2015
Lipase from the newly isolated Pseudomonas aeruginosa strain was produced, and its activity was enhanced through response surface methodology optimization. In the initial screening process, the bacterial strain was isolated from soil samples and then confirmed with lipase production on agar plates supplemented with Rhodamine B. The strain was identified by 16S rRNA gene sequence analysis. Plackett-Burman design was used to initially screen nine factors of culture medium. The design showed that four factors including peanut oil, sucrose, ammonium sulfate and volume of the medium exhibited significant affect on lipase production by P. aeruginosa strain. Afterward the optimal level of each significant factor was derived by Box-Behnken design and a 13.7 fold increase in lipase activity (21 U/mL-288 U/mL) was achieved under the optimum conditions. Thirty six percent of the soybean oil was converted into free fatty acids after hydrolyzing the oil for 12 h in the presence of optimized lipase.
Screening and Isolation of Lipase Producing Pseudomonas Sp. From Oil Contaminated Soil
International Research Journal Of Pharmacy, 2019
Pseudomonas sp. are found everywhere in the environment. Pseudomonas sp. are considered to be the most resistant and nuisance causing bacteria. Several Pseudomonas sp. have been shown to produce lipases. Leakages from pipelines, underground storage tanks of gas stations, improper disposal of petroleum wastes and stranded oil spills are the major sources of surface and groundwater contamination. The techniques used to remove hydrocarbons from such contaminated sites are quite expensive. In this study, we tried to find out the presence of oil degrading Pseudomonas sp. from oil contaminated sites. Various soil samples from oil contaminated sites were incubated in Pseudomonas sp. selective cetrimide broth. The enriched culture was screened for the presence of lipase producing organisms onto tributyrin agar. Further biochemical tests were performed for the confirmation of genus Pseudomonas.15 lipase producing microorganisms were obtained from 9 different soil samples. Isolate no-8 was found to be the highest lipase producing Pseudomonas sp. Presence of oil in the nutrient medium serves as a switch to produce lipase. As the enzyme lipase degrades oil and a fat, its production is higher in the organisms present in the oil contaminated soil. Lipase producing Pseudomonas sp. can be effectively used for the bioremediation of oil contaminated sites.
Enzymatic characterization of 30 kDa lipase from Pseudomonas aeruginosa ATCC 27853
Journal of Basic Microbiology, 2009
An extracellular lipase from Pseudomonas aeruginosa ATCC 27853 has been purified and its enzymatic characteristics were determined. According to SDS-PAGE and gel filtration molecular mass estimated to be 30 kDa, what classified the lipase in group I.1. Although 14 lipases from P. aeruginosa with similar molecular mass are referred to date, their basic enzymatic properties have not been reported yet. To address the gap we found: the optimal temperature and pH in water solution being 50 °C and 9.3, respectively; the lipase was inhibited with Hg 2+ ions and sodium dodecylsulphate (SDS), while non-ionic detergent Triton X-100 activated the enzyme; the lipase hydrolyzed more rapidly middle chain triglycerides and it was not regiospecific; the lipase demonstrated naturally occurring stability in different organic solvents with concentrations ranging from 30 to 70%, including good thermal stability in 30% organic solvent solution. Even though strain P. aeruginosa ATCC 27853 was not isolated from extreme environment it showed activity in organic solvent suggesting that this lipase is suitable for variety of applications, including reactions in water restricted medium and bioremediation of contaminations by organic solvents.
Characterization of a lipase from a newly isolated Pseudomonas sp
Iranian journal of microbiology, 2013
Lipases are valuable biocatalysts which are widely used in the detergent, food, dairy and pharmaceutical industries. The aims of the present study included the isolation of a lipase-producer from industrial zones and the partial characterization of the enzyme. A number of bacteria were isolated from sites related to the oil industries. An isolate forming a halo zone in a selective medium (TW agar) was then selected and grown on a medium suitable for the production of lipase. The isolate was subsequently identified by the 16S rRNA sequencing method, and its enzyme activity was measured by a spectrophotometer using pNPP as a substrate. The selected isolate was identified by the molecular method as Pseudomonas sp. Its extracellular lipase activity was 41.5 ± 1.4 U/ml, and the high affinity of this enzyme for the substrate was indicated by the kinetic parameters of Km and Vm, which were estimated by the the Lineweaver-Burk plot as 0.77 mM and 49.5 U/ml, respectively. Activation energy o...