Aqueous two Phase Systems for the Isolation and Partial Purification of Lipases from Soil Bacteria

Document Type: Research Article

Authors

1 University of Malakand

2 Department of Biotechnology, University of Malakand

3 Department of Biochemistry, Abdul Wali Khan University Mardan, Khyber Pakhtunkhwa, Pakistan

4 Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Khyber Pakhtunkhwa, Pakistan

5 Department of Microbiology, Kohat University of Science and Technology, Kohat, Khyber Pakhtunkhwa, Pakistan

Abstract

Aqueous two phase system (ATPS) is an effective, simple and single step technique used for the partial purification of biological molecules. The aim of the study was to develop ATPS technique for the isolation and partial purification of lipases. ATPS composed of polyethylene glycol and ammonium sulfate was used in this study to partially purify lipase from the fermentation broth of a bacillus strain isolated from local soil. The effect of different percent compositions of PEG 4000, 6000 and 10,000 along with 6.60%, 7.26%, 7.92% and 8.26% ammonium sulfate was studied on the separation behavior of lipase. The optimal condition for the isolation and partial purification of lipases from soil bacterial was 12.5 % of PEG 10,000 with 7.92% of ammonium sulfate. The recovery of lipase was 78.3% in the top phase (rich in PEG).

Keywords