Isolation, characterization and expression of genes encoding enzymes with lipase activity in Yarrowia lipolytica
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abstract
Yarrowia lipolytica is a dimophic yeast with very high secretory capacities and able to grow in
alkanes and fatty acids as the sole carbon source. Using PCR, we have isolated two DNA fragments
that after sequencing show homology with fungal lipases. By genomic DNA subcloning, we have
isolated a gene nominated YlLIP1 located on Y. lipolytica chromosome IV. The gene presents an ORF
of 1458 bp which encodes a putative protein of 486 amino acids with an apparent molecular weight of
55.4 kDa. The nucleotide sequence of the 5' and 3' ends shows the consensus signals for translation
initiation and transcription termination motifs in the yeast.
A disruption of YlLIP1 has been performed. The deleted ΔYllip1 strain shows
lower lipase activity in comparison to the wild type.
The DNA encoding YlLip1p was expressed in Saccharomyces cerevisiae and Kluyveromyces lactis.
Both yeast species are able to synthesize a functional enzyme under the control of GAL1 promoter.
We are currently isolating the second gene, both in the wild type and in the ΔYllip1 strain.
