Production of lipase from Pseudozyma aphidis and determination of the activity and stability of the crude lipase preparation in polar organic solvents

Abstract

The production of lipase from Pseudozyma aphidis (DSM 70725) was determined in six different media. The highest lipase production was observed in a medium with glucose as the sole carbon source, and yeast extract and sodium nitrate as the nitrogen sources. The time course studies of growth and lipase production in the optimal medium revealed that the highest lipase production was achieved at the end of the log phase of growth, reaching the value of 35.0 U cm -3 in the fifth day of cultivation. The effects of various polar, water-miscible, organic solvents on the activity and stability of the crude lipase produced by P. aphidis were evaluated. The hydrolytic activity of the crude lipase towards p-nitrophenyl palmitate (p-NPP) in aqueous media and in organic solvents was determined, using the same spectrophotometric assay in both the aqueous and organic media. The crude lipase preparation exhibited activity towards p-NPP only in acetone and acetonitrile, while the lipase was stable only in acetone, with 23 % residual activity after 24 h of incubation. These results suggested that lipase from P. aphidis can be used as a biocatalyst for potential applications in such organic solvents.

Activity and Stability of Lipase Preparations from Penicillium corylophilum: Potential Use in Biocatalysis

“…1). Lipases that are active and stable in polar solvents would open new possibilities and opportunities for biocatalysis involving polar substrates 22.…”

Purification and characterization of a thermo-halophilic, alkali-stable and extremely benzene tolerant esterase from a thermo-halo tolerant Bacillus cereus strain AGP-03, isolated from ‘Bakreshwar’ hot spring, India

“…It is assumed that enzymes from hot spring bacteria may be expected to possess bizarre properties and high enzyme stability in various adverse environmental/industrial conditions. Use of biocatalysts in organic solvents offers numerous advantages compared to traditional aqueous enzymology, such as ability to perform reactions restricted kinetically or thermodynamically in water, high solubility of certain substrates in organic solvents, suppression of undesirable side reactions, possibility to control or modify the enzyme selectivity, and possible elimination of microbial contamination [7][8][9]. Esterases are widely used due to their ability to catalyze not only the hydrolysis of triacylglycerides in aqueous solutions, but also enantio-selective synthetic reactions in organic media.…”

Design of biocompatible immobilized Candida rugosa lipase with potential application in food industry

“…Lipase activity was determined according to the modified hydrolytic assay with p ‐NPP. One unit of lipase activity (IU) was defined as the amount of enzyme that released 1 µmol p ‐nitrophenolate min −1 under the assay conditions …”

“…Stability and activity of CRL and HAP‐CRL in polar organic solvents . The method we used was described previously by Dimitrijevic et al …”

“…Stability and activity of CRL and HAP-CRL in polar organic solvents. The method we used was described previously by Dimitrijevic et al 25 Stability and activity of CRL and HAP-CRL in non-polar organic solvents. For stability assessment, 50 μL of enzyme preparation (free or immobilized form) was mixed with 950 μL of organic solvent and incubated for up to 3 h. At 0, 30 and 180 min, aliquots of the aqueous (enzyme) phase were collected and tested for hydrolytic lipase activity.…”