A novel lipase/acyltransferase from the yeast Candida albicans: expression and characterisation of the recombinant enzyme

Abstract

A gene encoding an extracellular lipase (CaLIP4) from Candida albicans was successfully expressed in Saccharomyces cerevisiae after mutagenesis of its unusual CUG serine codon into a universal one. The ability of this lipase, which shares 60% sequence homology with the lipase/acyltransferase from Candida parapsilosis, to synthesise esters was investigated. CaLIP4 behaved as a true lipase, displaying activity towards insoluble triglycerides and having no activity in the presence of short-chain fatty acid (FA) esters and phosphatidylcholine. Methyl, ethyl and propyl esters were efficiently used. The lipase exhibited highest selectivity for unsaturated FA. With saturated FAs, C14-C16 acyl chains were preferred. In a biphasic aqueous/lipid system, CaLIP4 displayed a high alcoholysis activity with a range of alcohols (e.g. methanol, ethanol, propanol and isopropanol) as acyl acceptor. During the course of the alcoholysis reaction, new esters are produced at concentrations above the thermodynamic equilibrium of the esterification reaction, indicating that ester synthesis does not proceed by esterification but mainly by direct acyltransfer. Ester synthesis is under kinetic control due to the high rate of alcoholysis. Unwanted hydrolysis is limited by competition between the acyl acceptor (alcohol) and water for the acyltransfer reaction, favouring the alcohol.

Expression, purification of a novel alkaline Staphylococcus xylosus lipase acting at high temperature

“…In addition to their hydrolytic activity, lipases can also catalyse other reactions such as esterification or interesterification. A particular interest relies on lipases capacity of catalysing such reactions and, consequently, the synthesis of fine compounds used for manufacturing products of high aggregate value, such as the engineering of structured lipids for the biotransformation of oils and fats [3][4][5][6]. Each application requires unique properties with respect to specificity, stability, optimal temperature, and pH-dependence [7].…”

Candida albicansCell Wall Proteins

“…LIP4 has been expressed in S. cerevisiae and the recombinant protein analyzed (320). The enzyme is a true lipase hydrolyzing insoluble triglycerides and is able to use methyl, ethyl, and propyl esters.…”

Yeast as an efficient biocatalyst for the production of lipid-derived flavours and fragrances

“…Although lipases possess a very conserved active site, there is a great diversity in the biocatalytic performances and new lipases are characterised regularly (to cite only very recent references: ; Roustan et al (2005)). It is particularly notable that the recently characterised Lip2p from Y. lipolytica has already given promising results with high enantioselectivity (E = 28) on the resolution of 2-bromo-p-tolylacetic acid ethyl ester (results comparable to the most efficient commercially available lipase (from Burkholderia cepacia: E = 30)) and E = 27 on the C. antartica resolution of 2-bromo-o-tolylacetic acid for which no commercial lipase was efficient (Guieysse et al 2004).…”