Browsing by Author "Burguillo, Francisco J."
Now showing 1 - 5 of 5
Results Per Page
Sort Options
- Isolation, characterization and expression of genes encoding enzymes with lipase activity in Yarrowia lipolyticaPublication . Choupina, Altino; Gonzalez, Francisco J.; Fermiñán, Encarnación; Burguillo, Francisco J.; Dominguez, ÁngelYarrowia 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.
- Lipase assay in yarrowia lipolyticaPublication . Morín, M.; Gonzalez, Francisco J.; Choupina, Altino; Burguillo, Francisco J.; Dominguez, ÁngelLipases (triacylglycerol acyl hydrolases, EC 3. 1.1.3) are key enzymes in fat metabolism produced by microorganisms, plants, and animals that catalyze the breakdown of triacylglycerols to free fatty adds and glycerol. In Y. lipolytica severaJ genes encoding lipases have been isoiated,LlP2 coding for a secreted lipase (Pignede et al. 1000) and LlPl and LIP3 encoding two lipase genes of the carboxylesrerase family (Choupina et al. 1999). Lipase expression was repressed by glucose and induced by fatty acids and olive oil.
- Non-conventional yeasts as hosts for heterologous protein production.Publication . Dominguez, Ángel; Fermiñán, Encarnación; Sánchez, Manuel; González, Francisco M.; Pérez-Campo, Flor Maria; García, Susana; Herrero, Ana B.; San Vicente, Avelino; Cabello, Juan; Prado, Marciano; Iglesias, Francisco J.; Choupina, Altino; Burguillo, Francisco J.; Fernández-Lago, Luis; López, Maria CarmenYeasts are an attractive group of lower eukaryotic microorganisms, some of which are used in several industrial processes that include brewing, baking and the production of a variety of biochemical compounds. More recently, yeasts have been developed as host organisms for the production of foreign (heterologous) proteins. Saccharomyces cerevisiae has usually been the yeast of choice, but an increasing number of alternative non-Saccharomyces yeasts has now become accessible for modern molecular genetics techniques. Some of them exhibit certain favourable traits such as high-level secretion or very strong and tightly regulated promoters, offering significant advantages over traditional bakers' yeast. In the present work, the current status of Kluyveromyces lactis, Yarrowia lipolytica, Hansenula polymorpha and Pichia pastoris (the best-known alternative yeast systems) is reviewed. The advantages and limitations of these systems are discussed in relation to S. cerevisiae.
- Producción de lipasas por la levadura Yarrowia lipolyticaPublication . Morín, M.; Choupina, Altino; Fermiñán, Encarnación; Burguillo, Francisco J.; Gonzalez, Francisco J.; Ruiz, L.; Dominguez, ÁngelYarrowia lipolytica es una levadura dimórfica que secreta gran cantidad de proteínas al medio de cultivo, entre ellas: proteasas - alcalinas, neutras y ácidas -, ribonucleasas, fosfatasas y lipasas. Las lipasas son enzimas importantes en el metabolismo de ácidos grasos, catalizando la hidrolisis de triglicéridos a ácidos grasos y glicerol.
- The lipase system of Yarrowia lipolyticaPublication . Choupina, Altino; Gonzalez, Francisco J.; Morín, M.; Burguillo, Francisco J.; Fermiñán, Encarnación; Dominguez, ÁngelAmong yeast species, Yarrowia lipolytica is one of the highest producers of extracellular proteins ( acid, neutral and alkaline proteases, ácid phosphatase, ribonucleases and lipases). Lipases ( triacylglycerol hydrolases) are important enzymes in fat metabolism, catalyzing the breakdown of triacilglycerols to free fatty acids and glycerol. Owing to the very low solubility of ther natural substrats, this hydrolysis is catalysed at the interfase beteween an insoluble substrat and the aqueous phase in which the enzyme is solubilized. This feature distinguishes them from esterases, which preferentially catalyze the hydrolisys of soluble esters in water.