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Applied Microbiology and Biotechnology

, Volume 68, Issue 2, pp 203–212 | Cite as

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

  • Jean Louis Roustan
  • Agustin Rascon Chu
  • Guy Moulin
  • Frédéric BigeyEmail author
Biotechnologically Relevant Enzymes and Proteins

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.

Keywords

Lipase Lipase Activity Fatty Acid Ethyl Ester Ester Synthesis Acyl Acceptor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This work was supported in part by Cognis Deutschland GmbH & Co. KG. We thank Johannes Crotogino and Ramses Obenga for their assistance in the cloning and mutagenesis experiments. We thank Professor Eric Dubreucq (UMR IR2B, ENSA.M.-INRA, Montpellier, France) for fruitful discussions throughout the research work and Guilhem Janbon (Institut Pasteur, Paris, France) for providing C. albicans DNA.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Jean Louis Roustan
    • 1
  • Agustin Rascon Chu
    • 1
  • Guy Moulin
    • 1
  • Frédéric Bigey
    • 1
    Email author
  1. 1.Unité Mixte de Recherche Ingénierie de la Réaction Biologique, BioproductionsInstitut National de la Recherche Agronomique-Ecole Nationale Supérieure Agronomique de MontpellierMontpellier cedex 1France

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