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Folia Microbiologica

, Volume 64, Issue 2, pp 245–255 | Cite as

Identification of cell-associated and secreted serine-type peptidases in multidrug-resistant emergent pathogens belonging to the Candida haemulonii complex

  • Xênia M. Souto
  • Lívia S. Ramos
  • Marta H. Branquinha
  • André L. S. SantosEmail author
Original Article

Abstract

The Candida haemulonii complex (Candida haemulonii, Candida haemulonii var. vulnera, and Candida duobushaemulonii) comprises emerging opportunistic human fungal pathogens with recognized multidrug-resistance profiles. Little is known about the virulence markers produced by this fungal complex. However, it is recognized that Candida spp. express a large array of peptidases, which play multiple roles in different aspects of fungal-host interactions. In the present study, we have identified proteolytic enzymes in clinical isolates of the C. haemulonii complex using zymographic assays. Peptidases able to hydrolyze gelatin, casein, albumin, hemoglobin, and immunoglobulin G were detected in cell-free supernatants and cellular extracts taken from the three species forming the C. haemulonii complex. Overall, peptidases were preferentially evidenced at physiological pH and temperatures of 37–42 °C, with molar masses between 35 and 85 kDa. Peptidase profiles of C. haemulonii and C. haemulonii var. vulnera isolates were quite similar, contrasting to the peptidases produced by C. duobushaemulonii. Almost all peptidases were inhibited by phenylmethanesulfonyl fluoride (PMSF), thus classifying them as serine-type peptidases. Additionally, proteolytic cleavage of soluble azoalbumin was blocked by PMSF (65–95% inhibition depending on the fungal isolate). These unprecedented results have demonstrated the capability of the C. haemulonii complex to produce serine-type peptidases with an ability to cleave a broad spectrum of proteins, including key host components.

Notes

Acknowledgments

The authors would like to thank Dr. Malachy McCann (Chemistry Department, National University of Ireland Maynooth, Co. Kildare, Ireland) for the valuable critical English review and Dr. Diogo de Azevedo Jurelevicius (Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Brazil) for the help with the dendrogram analysis.

Funding information

This study was supported by grants from Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2018

Authors and Affiliations

  1. 1.Laboratório de Estudos Avançados de Microrganismos Emergentes e Resistentes (LEAMER), Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes (IMPG)Universidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  2. 2.Programa de Pós-Graduação em Bioquímica, Instituto de Química (IQ)Universidade Federal do Rio de Janeiro (UFRJ)Rio de JaneiroBrazil

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