1887

Abstract

Extracellular phospholipases are demonstrated virulence factors for a number of pathogenic microbes. The opportunistic pathogen is known to secrete phospholipases and these have been correlated with strain virulence. In an attempt to clone genes encoding secreted phospholipases, was transformed with a genomic library and screened for lipolytic activity on egg-yolk agar plates, a traditional screen for phospholipase activity. Two identical clones were obtained which exhibited lipolytic activity. Nucleotide sequence analysis identified an ORF encoding a protein of 351 amino acid residues. Although no extensive homologies were identified, the sequence contained the Gly-X-Ser-X-Gly motif found in prokaryotic and eukaryotic lipases, suggesting a similar activity for the encoded protein. Indeed, culture supernatants from complemented yeast cells contained abundant hydrolytic activity against a triglyceride substrate and had no phospholipase activity. The data suggest that in addition to phospholipases, also has lipases. Southern blot analyses revealed that may contain a lipase gene () family, and that a lipase gene(s) may be present in and but not in or Northern blot analyses showed that expression of the transcript, the cloned gene which encodes a lipase, was detected only when was grown in media containing Tween 80, other Tweens or triglycerides as the sole carbon source, and not in Sabouraud Dextrose Broth or yeast/peptone/dextrose media. Additionally, carbohydrate supplementation inhibited expression. Cloning this gene will allow the construction of -deficient null mutants which will be critical in determining the role of this gene in candidal virulence.

Keyword(s): Candida albicans , LIP1 and lipase
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/content/journal/micro/10.1099/00221287-143-2-331
1997-02-01
2021-08-01
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