1887

Abstract

is a dimorphic fungus and a member of the normal cutaneous microflora of humans. However, it is also a facultative pathogen, associated with a wide range of skin diseases. One unusual feature of is an absolute dependency on externally provided lipids which the fungus hydrolyses by lipolytic activity to release fatty acids necessary for both growth and pathogenicity. In this study, the cloning and characterization of the first gene encoding a secreted lipase of possibly associated with this activity are reported. The gene, , shows high sequence similarity to other known extracellular lipases, but is not a member of a lipase gene family in consists of 1464 bp, encoding a protein with a molecular mass of 54·3 kDa, a conserved lipase motif and an N-terminal signal peptide of 26 aa. By using a genomic library, two other genes were identified flanking , one of them encoding a putative secreted catalase, the other a putative amine oxidase. The cDNA of was expressed in and the biochemical properties of the recombinant lipase were analysed. MfLip1 is most active at 40 °C and the pH optimum was found to be 5·8. The lipase hydrolysed lipids, such as Tweens, frequently used as the source of fatty acids in media, and had minor esterase activity. Furthermore, the lipase is inhibited by different bivalent metal ions. This is the first molecular description of a secreted lipase from .

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28501-0
2006-02-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/152/2/547.html?itemId=/content/journal/micro/10.1099/mic.0.28501-0&mimeType=html&fmt=ahah

References

  1. Altschul S. F, Gish W, Miller W, Myers E. W, Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410 [CrossRef]
    [Google Scholar]
  2. Ashbee H. R, Evans E. G. 2002; Immunology of diseases associated with Malassezia species. Clin Microbiol Rev 15:21–57 [CrossRef]
    [Google Scholar]
  3. Bendtsen J. D, Nielsen H, Brunak S, von Heijne G. 2004; Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340:783–795 [CrossRef]
    [Google Scholar]
  4. Biswas S. K, Yokoyama K, Nishimura K, Miyaji M. 2001; Molecular phylogenetics of the genus Rhodotorula and related basidiomycetous yeasts inferred from the mitochondrial cytochrome b gene. Int J Syst Evol Microbiol 51:1191–1199 [CrossRef]
    [Google Scholar]
  5. Bojar R. A, Tue C. J, Holland K. T. 2004; The effect of lipids on the adherence of axillary aerobic coryneform bacteria. Lett Appl Microbiol 38:470–475 [CrossRef]
    [Google Scholar]
  6. Caprilli F, Mercantini R, Nazzaro Porro M, Passi S, Tonolo A. 1973; Studies of the genus Pityrosporum in submerged culture. Mycopathol Mycol Appl 51:171–189 [CrossRef]
    [Google Scholar]
  7. Catterall M. D, Ward M. E, Jacobs P. 1978; A reappraisal of the role of Pityrosporum orbiculare in pityriasis versicolor and the significance of extracellular lipase. J Invest Dermatol 71:398–401 [CrossRef]
    [Google Scholar]
  8. Crespo Erchiga V, Delgado Florencio V. 2002; Malassezia species in skin diseases. Curr Opin Infect Dis 15:133–142 [CrossRef]
    [Google Scholar]
  9. Derewenda Z. S. 1994; Structure and function of lipases. Adv Protein Chem 45:1–52
    [Google Scholar]
  10. Derewenda Z. S, Derewenda U. 1991; Relationships among serine hydrolases: evidence for a common structural motif in triacylglyceride lipases and esterases. Biochem Cell Biol 69:842–851 [CrossRef]
    [Google Scholar]
  11. Eisenhaber B, Bork P, Eisenhaber F. 1998; Sequence properties of GPI-anchored proteins near the omega-site: constraints for the polypeptide binding site of the putative transamidase. Protein Eng 11:1155–1161 [CrossRef]
    [Google Scholar]
  12. Eisenhaber B, Schneider G, Wildpaner M, Eisenhaber F. 2004; A sensitive predictor for potential GPI lipid modification sites in fungal protein sequences and its application to genome-wide studies for Aspergillus nidulans , Candida albicans , Neurospora crassa , Saccharomyces cerevisiae and Schizosaccharomyces pombe . J Mol Biol 337:243–253 [CrossRef]
    [Google Scholar]
  13. Ellerbroek P. M, Walenkamp A. M, Hoepelman A. I, Coenjaert F. E. 2004; Effects of the capsular polysaccharides of Cryptococcus neoformans on phagocyte migration and inflammatory mediators. Curr Med Chem 11:253–266 [CrossRef]
    [Google Scholar]
  14. Emanuelsson O, Nielsen H, Brunak S, von Heijne G. 2000; Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016 [CrossRef]
    [Google Scholar]
  15. Faergemann J, Aly R, Maibach H. I. 1983; Adherence of Pityrosporum orbiculare to human stratum corneum cells. Arch Dermatol Res 275:246–250
    [Google Scholar]
  16. Gueho E, Midgley G, Guillot J. 1996; The genus Malassezia with description of four new species. Antonie Van Leeuwenhoek 69:337–355 [CrossRef]
    [Google Scholar]
  17. Gueho E, Boekhout T, Ashbee H. R, Guillot J, Van Belkum A, Faergemann J. 1998; The role of Malassezia species in the ecology of human skin and as pathogens. Med Mycol 36:Suppl 1220–229
    [Google Scholar]
  18. Guillot J, Lesourd M, Midgley G, Dupont B, Guého E, Chévrier G. 1996; Identification of Malassezia species. A practical approach. J Mycol Med 6:103–110
    [Google Scholar]
  19. Gupta A. K, Kohli Y, Summerbell R. C. 2000; Molecular differentiation of seven Malassezia species. J Clin Microbiol 38:1869–1875
    [Google Scholar]
  20. Gupta A. K, Batra R, Bluhm R, Boekhout T, Dawson T. L. Jr 2004; Skin diseases associated with Malassezia species. J Am Acad Dermatol 51:785–798 [CrossRef]
    [Google Scholar]
  21. Hammer K. A, Riley T. V. 2000; Precipitate production by some Malassezia species on Dixon's agar. Med Mycol 38:105–107 [CrossRef]
    [Google Scholar]
  22. Hoffmann G. E, Schmidt D, Bastian B, Guder W. G. 1986; Photometric determination of phospholipase A. J Clin Chem Clin Biochem 24:871–875
    [Google Scholar]
  23. Hube B, Stehr F, Bossenz M, Mazur A, Kretschmar M, Schafer W. 2000; Secreted lipases of Candida albicans : cloning, characterisation and expression analysis of a new gene family with at least ten members. Arch Microbiol 174:362–374 [CrossRef]
    [Google Scholar]
  24. Kano R, Aizawa T, Nakamura Y, Watanabe S, Hasegawa A. 1999; Chitin synthase 2 gene sequence of Malassezia species. Microbiol Immunol 43:813–815 [CrossRef]
    [Google Scholar]
  25. Kesavan S, Holland K. T, Ingham E. 2000; The effects of lipid extraction on the immunomodulatory activity of Malassezia species in vitro . Med Mycol 38:239–247 [CrossRef]
    [Google Scholar]
  26. Krogh A, Larsson B, Sonnhammer E. L, von Heijne G. 2001; Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes. J Mol Biol 305:567–580 [CrossRef]
    [Google Scholar]
  27. Lee S. A, Wormsley S, Kamoun S, Lee A. F, Joiner K, Wong B. 2003; An analysis of the Candida albicans genome database for soluble secreted proteins using computer-based prediction algorithms. Yeast 20:595–610 [CrossRef]
    [Google Scholar]
  28. Lu Z, Szafron D, Greiner R, Lu P, Wishart D. S, Poulin B, Anvik J, Macdonell C, Eisner R. 2004; Predicting subcellular localization of proteins using machine-learned classifiers. Bioinformatics 20:547–556 [CrossRef]
    [Google Scholar]
  29. Mancianti F, Rum A, Nardoni S, Corazza M. 2001; Extracellular enzymatic activity of Malassezia spp. isolates. Mycopathologia 149:131–135 [CrossRef]
    [Google Scholar]
  30. Mayser P, Haze P, Papavassilis C, Pickel M, Gruender K, Gueho E. 1997; Differentiation of Malassezia species: selectivity of cremophor EL, castor oil and ricinoleic acid for M. furfur . Br J Dermatol 137:208–213 [CrossRef]
    [Google Scholar]
  31. Mittag H. 1995; Fine structural investigation of Malassezia furfur . II. The envelope of the yeast cells. Mycoses 38:13–21 [CrossRef]
    [Google Scholar]
  32. Plotkin L. I, Squiquera L, Mathov I, Galimberti R, Leoni J. 1996; Characterization of the lipase activity of Malassezia furfur . J Med Vet Mycol 34:43–48 [CrossRef]
    [Google Scholar]
  33. Porro M. N, Passi S, Caprill F, Nazzaro P, Morpurgo G. 1976; Growth requirements and lipid metabolism of Pityrosporum orbiculare . J Invest Dermatol 66:178–182 [CrossRef]
    [Google Scholar]
  34. Ran Y, Yoshiike T, Ogawa H. 1993; Lipase of Malassezia furfur : some properties and their relationship to cell growth. J Med Vet Mycol 31:77–85 [CrossRef]
    [Google Scholar]
  35. Sambrook J, Russell D. 2000 Molecular Cloning: a Laboratory Manual, 3rd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  36. Sansinforiano M. E, Padilla J. A, Fernandez-Garcia J. L, Martinez-Trancon M, Rabasco A, Parejo J. C, Hermoso de Mendoza J, Hermoso de Mendoza M. 1998; Rapid and easy method to extract and preserve DNA from Cryptococcus neoformans and other pathogenic yeasts. Mycoses 41:195–198 [CrossRef]
    [Google Scholar]
  37. Shifrine M, Marr A. G. 1963; The requirement of fatty acids by Pityrosporum ovale . J Gen Microbiol 32:263–270 [CrossRef]
    [Google Scholar]
  38. Xie D, Li A, Wang M, Fan Z, Feng H. 2005; locsvmpsi: a web server for subcellular localization of eukaryotic proteins using svm and profile of psi-blast. Nucleic Acids Res 33:W105–W110 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28501-0
Loading
/content/journal/micro/10.1099/mic.0.28501-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error