Single-step purification of lipase from Burkholderia multivorans using polypropylene matrix

Lipase from Burkholderia multivorans was purified with high yields directly from fermentation broth by a single-step purification protocol involving adsorption and desorption. The crude enzyme ( lyophilized powder) from B. multivorans was loaded on Accurel ( Membrana, Germany), a polypropylene matrix, using butanol as the solvent in a buffer at pH 9.0 and ambient temperature for a period of 12 h. The enzyme adsorbed onto the matrix with high specific activity ( 33 units mg(-1) protein). This was followed by desorption of the enzyme from the matrix using Triton X-100 as the eluent. The enzyme was finally recovered by precipitation with acetone (50%, v/v). Thus, an overall enzyme yield of 66% with a 3.0-fold purification was obtained. The purity of the enzyme was ascertained by SDS-PAGE. The phenomenon of adsorption and desorption on Accurel was studied for three more lipases, viz. Mucor meihei lipase ( Sigma - Aldrich Co.), Lipolase ( Novo Nordisk, Denmark) and Pseudomonas aeruginosa lipase ( laboratory isolate).