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).
Gupta
Namita
N
Department of Microbiology, University of Delhi, South Campus, Benito Juarez Road, New Delhi 110021, India.
Rathi
Pooja
P
Singh
Rajni
R
Goswami
Vineet Kumar
VK
Gupta
Rani
R
D020001:1-Butanol; D000096:Acetone; D000327:Adsorption; D019117:Burkholderia / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005590:Fractional Precipitation; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000302:isolation & purification*; D009089:Mucor / Q000201:enzymology; D017830:Octoxynol; D011126:Polypropylenes; D011549:Pseudomonas / Q000201:enzymology; D013997:Time Factors
D011126:Polypropylenes; D000096:Acetone; D020001:1-Butanol; D017830:Octoxynol; D008049:Lipase