Identification and characterisation of LIP7 and LIP8 genes encoding two extracellular triacylglycerol lipases in the yeast Yarrowia lipolytica.
In the lipolytic yeast Yarrowia lipolytica, the LIP2 gene was previously reported to encode an extracellular lipase. The growth of a Deltalip2 strain on triglycerides as sole carbon source suggest an alternative pathway for triglycerides utilisation in this yeast. Here, we describe the isolation and the characterisation of the LIP7 and LIP8 genes which were found to encode a 366 and a 371-amino acid precursor protein, respectively. These proteins which belong to the triacylglycerol hydrolase family (EC 3.1.1.3) presented a high homology with the extracellular lipase CdLIP2 and CdLIP3 from Candida deformans. The physiological function of the lipase isoenzymes was investigated by creating single and multi-disrupted strains. Lip7p and Lip8p were found to correspond to active secreted lipases. The lack of lipase production in a Deltalip2 Deltalip7 Deltalip8 strain suggest that no additional extracellular lipase remains to be discovered in Y. lipolytica. The substrate specificity towards synthetic ester molecules indicates that Lip7p presented a maximum activity centred on caproate (C6) while that of Lip8p is in caprate (C10).
Fickers
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P
Laboratoire Microbiologie et Génétique Moléculaire, UMR2585 CNRS INRA INAP-G Institut National Agronomique Paris-Grignon, F-78850 Thiverval-Grignon, France.
Fudalej
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F
Le Dall
M T
MT
Casaregola
S
S
Gaillardin
C
C
Thonart
P
P
Nicaud
J M
JM
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D005658:Fungi / Q000235:genetics; D017353:Gene Deletion; D005838:Genotype; D008049:Lipase / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D025062:Yarrowia / Q000201:enzymology / Q000235:genetics*
D008049:Lipase