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Characterization of a lipase from a newly isolated Pseudomonas sp.

Noormohamadi R, Tabandeh F, Shariati P, Otadi M - Iran J Microbiol (2013)

Bottom Line: 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 of lipase calculated from the Arrhenius plot was found to be 20.78 kJ/mol, and a temperature coefficient (Q10) of 4.39 indicated the high catalytic activity of the enzyme and the temperature dependence of the enzymatic reaction.The results demonstrated that the indigenous isolate could have potential applications in many relevant industries.

View Article: PubMed Central - PubMed

Affiliation: Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran ; Department of Chemical Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran.

ABSTRACT

Background and objectives: 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.

Materials and methods: 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.

Results: 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 of lipase calculated from the Arrhenius plot was found to be 20.78 kJ/mol, and a temperature coefficient (Q10) of 4.39 indicated the high catalytic activity of the enzyme and the temperature dependence of the enzymatic reaction.

Conclusion: The results demonstrated that the indigenous isolate could have potential applications in many relevant industries.

No MeSH data available.


Related in: MedlinePlus

The vertical axis is enzyme activity, the horizontal axis shows the duration of incubation for 10, 20, 30, 40, 50 and 60 min. The colum from left to right demonstrate the decrease in activity of the enzyme at 30, 40, 50, 60, 70, 80 and 90 OC, during incubation.
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Figure 4: The vertical axis is enzyme activity, the horizontal axis shows the duration of incubation for 10, 20, 30, 40, 50 and 60 min. The colum from left to right demonstrate the decrease in activity of the enzyme at 30, 40, 50, 60, 70, 80 and 90 OC, during incubation.

Mentions: The residual activities of lipase at different pH values and temperatures were measured during 60 min so as to determine the optimum pH and thermo-stability of the enzyme (Fig. 4). The results showed that 68% of lipase activity was present after 30 min of incubation at 40°C and pH 8. The lipase lost 32% and 94% activity when it was incubated at pH 8 for 30 and 60 min, at 30°C, respectively. The enzyme was active only for 10 min at 90°C. The residual activities of 18.1 ± 1.5, 20.9 ± 0.8, 20.9 ± 1.1, 30.9 ± 1.0, 15.2 ± 2.1, 6.5 ± 0.7, 0 U/ml were obtained after 20 min of incubation at 30, 40, 50, 60, 70, 80, and 90°C, respectively.


Characterization of a lipase from a newly isolated Pseudomonas sp.

Noormohamadi R, Tabandeh F, Shariati P, Otadi M - Iran J Microbiol (2013)

The vertical axis is enzyme activity, the horizontal axis shows the duration of incubation for 10, 20, 30, 40, 50 and 60 min. The colum from left to right demonstrate the decrease in activity of the enzyme at 30, 40, 50, 60, 70, 80 and 90 OC, during incubation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4385172&req=5

Figure 4: The vertical axis is enzyme activity, the horizontal axis shows the duration of incubation for 10, 20, 30, 40, 50 and 60 min. The colum from left to right demonstrate the decrease in activity of the enzyme at 30, 40, 50, 60, 70, 80 and 90 OC, during incubation.
Mentions: The residual activities of lipase at different pH values and temperatures were measured during 60 min so as to determine the optimum pH and thermo-stability of the enzyme (Fig. 4). The results showed that 68% of lipase activity was present after 30 min of incubation at 40°C and pH 8. The lipase lost 32% and 94% activity when it was incubated at pH 8 for 30 and 60 min, at 30°C, respectively. The enzyme was active only for 10 min at 90°C. The residual activities of 18.1 ± 1.5, 20.9 ± 0.8, 20.9 ± 1.1, 30.9 ± 1.0, 15.2 ± 2.1, 6.5 ± 0.7, 0 U/ml were obtained after 20 min of incubation at 30, 40, 50, 60, 70, 80, and 90°C, respectively.

Bottom Line: 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 of lipase calculated from the Arrhenius plot was found to be 20.78 kJ/mol, and a temperature coefficient (Q10) of 4.39 indicated the high catalytic activity of the enzyme and the temperature dependence of the enzymatic reaction.The results demonstrated that the indigenous isolate could have potential applications in many relevant industries.

View Article: PubMed Central - PubMed

Affiliation: Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran ; Department of Chemical Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran.

ABSTRACT

Background and objectives: 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.

Materials and methods: 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.

Results: 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 of lipase calculated from the Arrhenius plot was found to be 20.78 kJ/mol, and a temperature coefficient (Q10) of 4.39 indicated the high catalytic activity of the enzyme and the temperature dependence of the enzymatic reaction.

Conclusion: The results demonstrated that the indigenous isolate could have potential applications in many relevant industries.

No MeSH data available.


Related in: MedlinePlus