New Colorimetric Method for Lipases Activity Assay in Microbial Media

Mohamed A. Abd-Elhakeem, Ahmed M. Elsayed, Taher A. Alkhulaqi

College of Biotechnology, Misr University for Science and Technology, Giza, Egypt.
Research and Development Center, Misr University for Science and Technology, Giza, Egypt.

DOI: 10.4236/ajac.2013.49055                

Abstract

A simple, rapid and precise method has been developed for determination of lipase activity in microbial media. The method is based on using phenyl acetate as substrate for lipase and determination of liberated phenol by Folin Ciocalteu reagent. Reaction mixture containing substrate 2.4 ml of phenyl acetate 165 μM in Tris HCl buffer, 0.1 M and pH 7, with 1% (v/v) Triton X-100) and 0.1 ml lipase is incubated at 40?C during 10 minutes and the absorbance was measured at 750 nm. Linearity was observed in the concentration range 0-0.8 g/L lipase.

Keywords

Lipase; Colorimetric Method; Assay; Phenyl Acetate; Folin

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	R. Sharma, Y. Chisti and U. C. Banerjee, “Production, Purification, Characterization, and Applications of Lipases,” Biotechnology Advances, Vol. 19, No. 8, 2001, pp. 627-662. doi:10.1016/S0734-9750(01)00086-6
[2]	K. R. Shah and S. A. J. Bhatt, “Purification and Characterization of Lipase from Bacillus subtilis Pa2,” Journal of Biochemical Technology, Vol. 3, No. 3, 2011, pp. 292-295.
[3]	F. Beisson, A. Tiss, C. Rivi_ere and R. Verger, “Methods for Lipase Detection and Assay: a Critical Review,” Eur. European Journal of Lipid Science and Technology, Vol. 102, No. 2, 2000, pp. 133-115. doi:10.1002/(SICI)1438-9312(200002)102:2<133::AID-EJLT133>3.3.CO;2-O
[4]	F. Hasan, A. A. Shah and A. Hameed, “Methods for Detection and Characterization of Lipases: A Comprehensive Review,” Biotechnology Advances, Vol. 27, No. 6, 2009, pp. 782-798. doi:10.1016/j.biotechadv.2009.06.001
[5]	A. A. Amara and S. R. Salem, “Degradation of Castor Oil and Lipase Production by Pseudomonas aeruginosa,” American-Eurasian Journal of Agricultural & Environmental Sciences, Vol. 5, No. 4, 2009, pp. 556-563.
[6]	D.-S. Park, H.-W. Oh, S.-Y. Heo, W.-J. Jeong, D. H. Shin, K. S. Bae and H.-Y. Park, “Characterization of an Extracellular Lipase in Burkholderiasp. HY-10 Isolated from a Longicorn Beetle,” The Journal of Microbiology, Vol. 45, No. 5, 2007, pp. 409-417.
[7]	D. R. Lewis, D. Zhou and J. Liu, “Direct Measurement of Lipase Inhibition by Orlistat Using a Dissolution Linked In Vitro Assay,” International Journal of Clinical Pharmacology and Biopharmacy, Vol. 1, No. 3, 2012, pp. 1-3.
[8]	Shao-Hua Chiou, Wen-Teng Wu. “Immobilization of Candida rugosa Lipase on Chitosan with Activation of the Hydroxyl Groups,” Biomaterials, Vol. 25, No. 2, 2004, pp. 197-204. doi:10.1016/S0142-9612(03)00482-4
[9]	N. Gomes, C. Goncalves, M. Garcia-Roman, J. A. Teixeiraa and I. Belo. “Optimization of a Colorimetric Assay for Yeast Lipase Activity in Complex Systems,” Analytical Methods, Vol. 3, No. 4, 2011, pp. 1008-1013. doi:10.1039/c0ay00680g
[10]	N. Gupta, P. Rathi and R. Gupta. “Simplified Para-Nitrophenyl Palmitate Assay for Lipases and Esterases,” Analytical Biochemistry, Vol. 311, No. 1, 2002, pp. 98-99. doi:10.1016/S0003-2697(02)00379-2