[en] The yeast Yarrowia lipolytica produces an extracellular lipase encoded by the LIP2 gene. However, very little is known about the mechanisms controlling its expression, especially on glucose media. In this work, the involvement of hexokinase Hxk1 in the glucose catabolite repression of LIP2 was investigated in a lipase overproducing mutant less sensitive to glucose repression. This mutant has a reduced capacity to phosphorylate hexose compared with the wild-type strain, but no differences could be observed between the HXK1 sequences in the two isolates. This suggested that the reduced phosphorylating activity of the mutant strain probably resulted from a modification in the level of HXK1 expression. However, overexpression of the HXK1 gene in this mutant led to a decrease of both LIP2 induction and extracellular lipase activity, suggesting that the hexokinase is involved in the glucose catabolite repression of LIP2 in Y lipolytica.
G Barth C Gaillardin 1997 Physiology and genetics of the dimorphic fungus Yarrowia lipolytica FEMS Microbiol Rev 19 219 237
J Destain D Roblain P Thonart 1997 Improvement of lipase production from Y lipolytica Biotechnol Lett 19 105 107
A Does L Bisson 1989 Comparison of glucose uptake kinetics in different yeasts J Bacteriol Lett 171 1303 1308
P Fickers JM Nicaud T Destain P Thonart 2003 Over-production of lipase by Yarrowia lipolytica mutants Appl Microbiol Biotechnol 63 136 142
C Gaillardin AM Ribet 1987 LEU2 directed expression of beta-galactosidase activity and phleomycin resistance in Yarrowia lipolytica Curr Genet 11 369 375
JM Gancedo D Clifton D Fraenkel 1977 Yeast hexokinase mutants J Biol Chem 252 4443 4444
M Herrero C Martinez-Campa F Moreno 1998 The hexokinase 2 protein participates in regulatory DNA-protein complexes necessary for glucose repression of the SUC2 gene in Saccharomyces Cerevisiae FEBS Lett 434 71 76
MT Le Dall JM Nicaud C Gaillardin 1994 Multiple-copy integration in the yeast Yarrowia lipolytica Curr Genet 26 38 44
C Madzak B Treton S Blanchin-Roland 2000 Strong hybrid promoters and integrative expression/secretion vectors for quasi-constitutive expression of heterologous proteins in the yeast Yarrowia lipolytica J Mol Microbiol Biotechno 2 207 216
M Maftahi C Gaillardin JM Nicaud 1996 Sticky-end polymerase chain reaction method for systematic gene disruption in Saccharomyces cerevisiae Yeast 12 859 868
T Petit C Gancedo 1999 Molecular cloning and characterization of the gene HXK1 encoding the hexokinase from Yarrowia lipolytica Yeast 15 1573 1584
G Pignede W Wang F Fudalej C Gaillardin M Seman JM Nicaud 2000 Characterisation of an extracellular lipase encoded by LIP2 in Yarrowia lipolytioca J Bacteriol 182 2802 2810
L Sarda P Desnuelle 1958 Action de la lipase pancreatique sur les ester en emulsion Biochim Biophys Acta 30 513 521
J Sambrook EF Fritsch T Maniatis 1989 Molecular cloning: a laboratory manual Cold Spring Harbor Laboratory Cold Spring Harbor, NY