World Journal of
Pharmaceutical and Life Sciences

PURIFICATION AND CHARACTERIZATION OF LIPASE FROM PSEUDOMONAS AERUGINOSA SJ2

Joshi Swapnil Satish* and Jobanputra Arpana Hemraj

ABSTRACT

The present study was aimed to purify and characterize extracellular lipase from a newly isolated Pseudomonas aeruginosa SJ2. An extracellular lipase with molecular weight of 32,768 Da was purified using salt precipitation, ion exchange chromatography and molecular exclusion chromatography. Purification resulted in 2,826 U/mg specific activity with 6.57-fold purification and 74.24% yield. SDS-PAGE and revealed it to be near to homogeneity showing a relative molecular weight of about 32,768 Da that was confirmed by mass spectrometry. Zymography confirmed the activity of the purified lipase. Further stability studies were performed on the purified lipase. The optimum temperature and pH for activity were found to be 40°C and pH 8.0. Ca2+ ions at 1mM stimulated lipase activity while other metals inhibited lipase activity. Lipase retained good residual activity in the presence of various organic solvents (methanol, iso-propanol, ethanol, toluene, n-hexane, DMSO, and acetone). Lipase was fairly stable in the presence of non-ionic surfactants while lost some of its activity in the presence of ionic surfactants. Lipase was fairly stable in the presence of inhibitors (EDTA, urea, PMSF, ?-mercaptoethanol). For oxidizing agents lipase was fairly stable in the presence of NaOCl and H2O2 while some of its activity was lost in the presence of NaBO3. Lipase was fairly stable and retained 92.15 % and 76.67 % relative activity after 30 days of storage at 4°C and 30°C respectively. The Km and Vmax were 0.24 mM and 3.51 ?mol/min/mg respectively towards pNPP as a substrate.

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