Isolation, characterization and expression of genes encoding enzymes with lipas...

• Author(s): Choupina, Altino ; Gonzalez, Francisco J. ; Fermiñán, E. ; Burguillo, F.J. ; Domínguez, A.
• Date: 1998
• Persistent ID: http://hdl.handle.net/10198/10942
• Origin: Biblioteca Digital do IPB
• Subject(s): Yeast; Lipases
• Document Type: Conference Object
• Language: English

Description

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.