Identification of a triacylglycerol lipase gene family in Candida deformans: molecular cloning and functional expression - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Yeast Année : 2003

Identification of a triacylglycerol lipase gene family in Candida deformans: molecular cloning and functional expression

Résumé

The yeast Candida deformans CBS 2071 produces an extracellular lipase which was shown to catalyse the production of various esters by the esterification of free fatty acids, even in the presence of a large molar excess of water. To clone the gene encoding this extracellular lipase, Saccharomyces cerevisiae was transformed with C deformans genomic libraries and screened for lipolytic activity on a medium containing rapeseed oil emulsion and rhodamine B. Three members of a lipase gene family (CdLIP1, CdLIP2 and CdLIP3) were cloned and characterized. Each deduced lipase sequence has a Gly-His-Ser-Leu-Gly-(Gly/Ala)-Ala conserved motif, eight cysteine residues and encodes an N-terminal signal sequence. MALDI-TOF mass spectrometry analysis of a proteolytic digest of the lipase produced was used to obtain experimental evidence that the CdLIP1 gene encoded the extracellular lipase. Recombinant expression studies confirmed that the cloned genes encoded functional lipases. The three lipases are very similar to lipases from the related species Yarrowia lipolytica. Significant homologies were also found with several yeast and fungal lipases. As C. deformans CBS 2071 was previously considered to be synonymous with Y. lipolytica, the strains were compared for the extent of nucleotide divergence in the variable regions (D1/D2) at the F-end of the large-subunit (26S) ribosomal DNA (rDNA) gene. This rDNA region has diverged sufficiently to suggest that C. deformans is a separate species. The nucleotide sequences of the CdLIP1, CdLIP2 and CdLIP3 genes will appear in the EMBL nucleotide sequence database under Accession Nos AJ428393, AJ428394 and AJ428395, respectively.
Fichier principal
Vignette du fichier
2003_Bigey_Yeast.pdf (188.03 Ko) Télécharger le fichier
Origine : Accord explicite pour ce dépôt
Loading...

Dates et versions

hal-02165415 , version 1 (25-06-2019)

Licence

Paternité

Identifiants

Citer

Frederic Bigey, Karine Tuery, Daisy Bougard, Jean-Marc Nicaud, Guy Moulin. Identification of a triacylglycerol lipase gene family in Candida deformans: molecular cloning and functional expression. Yeast, 2003, 20 (3), pp.233-248. ⟨10.1002/yea.958⟩. ⟨hal-02165415⟩
22 Consultations
311 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More