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Partitioning Optimization of Lipase from Burkholderia cepacia by PEG 1500/Phosphate Aqueous Two-Phase System

Padilha GS, Santana JCC, Alegre RM and Tambourgi EB

This work intended to purify an extracellular lipase from Burkholdera cepacia (ATCC 25416) by aqueous liquid-liquid two-phase using polyethylene glycol 1500 Da and salt phosphate. An experimental planning was done to evaluate the effects of pH (6.0, 7.0 and 8.0) and tie lines lengths (PEG and salt balance concentrations) on the partition coefficient. Lipases were obtained from an environment containing soy oil by fermentation. RSM optimization results show that the enzyme partition coefficient depended on pH and tie line, where the best resultswere found at pH 6 and tie lines 3, in this case, in relationship the crude extract, the purification factor was amongst 3 times higher.While on the protein partition, the best result was in pH 8.0 and tie line 3.After theATPS process, the top phase sample of the best systemwas analyzed by SDS–PAGE. The molecular weight was determined as 33 kDa using a polyacrylamide gel under denaturing conditions.