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An extracellular thermostable lipase from Amycolatopsis mediterranei DSM 43304 has been purified to homogeneity using ammonium sulphate precipitation followed by anion exchange chromatography and hydrophobic interaction chromatography. This protocol resulted in 398 fold purification with 36% final recovery. The purified A. mediterranei DSM 43304 lipase (AML) has an apparent molecular mass of 33 kDa. The N-terminal sequence, AANPYERGPDPTTASIEATR, showed highest similarity to a lipase from Streptomyces exfoliatus. The values of and for p-nitrophenyl palmitate (p-NPP) under optimal temperature (60°C) and pH (8.0) conditions were 0.10 ± 0.01 mM and 2.53 ± 0.06 mmol/minmg, respectively. The purified AML displayed significant activity towards a range of short and long chain triglycerides. It was most active on triolein and a wide range of p-nitrophenyl esters, with a preference for an acyl chain length of C8:0. Hydrolysis of glycerol esterbonds occurred non-specifically. The purified AML displayed significant stability in the presence of organic solvents (40% v/v) and catalyzed the synthesis of flavour ester isoamyl acetate in free and immobilized states.
Dheeman, D. S., Henehan, G.T.M. and Frias, J.M. (2011) Purification and properties of Amycolatopsis mediterranei DSM 43304 lipase and its potential in flavour ester synthesis. Bioresource Technologies 102(3) 3373-3379. doiL10.1016/j.biortech.2010.11.074