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

Single-step purification of lipase from Burkholderia multivorans using polypropylene matrix
February 2005
Gupta N, Rathi P, Singh R, Goswami VK, Gupta R.,
Department of Microbiology, University of Delhi, South Campus, Benito Juarez Road, New Delhi, 110021, India
Appl Microbiol Biotechnol. 2005 Feb 12; [Epub ahead of print]
Received: 2 September 2004 Revised: 9 November 2004 Accepted: 22 November 2004 Published online: 12 February 2005
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Abstract
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).