Distinct chitinases are expressed during various growth phases of the human pathogen Paracoccidioides brasiliensis
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2012-05
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The aim of this work was the partial purification and subsequent evaluation of chitinase expression during the
various growth phases of Paracoccidioides brasiliensis. Initially, PbCTS1r was expressed as a recombinant pro-
tein and displayed enzymatic activity against 4-MU-[N-acetylglucosamine (GlcNAc)] 3 and 4-MU-(GlcNAc) 2 . Two
proteins, 45 kDa and 39 kDa in size, were partially purified from P. brasiliensis yeast crude extract using cation-
exchange chromatography coupled with HPLC and were characterised as PbCTS1 and PbCTS2, respectively. Anti-
PbCTS1r antibody recognised two proteins in the crude extracts of yeast and the transitional stage between mycelial
and yeast phases. In crude extracts of mycelium, only the 45 kDa protein was detected. However, quantitative real-
time polymerase chain reaction led to the detection of small quantities of Pbcts2 transcript in the mycelial phase. In
the yeast cell wall extract, only the 39 kDa protein was detected. Moreover, both proteins were secreted by the yeast
parasitic phase, suggesting that these proteins participate in the modulation of the fungal environment. Phylogenetic
analysis of the predicted PbCTS1 and PbCTS2 proteins indicated that they code for distinct chitinases in P. brasilien-
sis. During evolution, P. brasiliensis could have acquired the paralogues Pbcts1 and Pbcts2 for growth and survival
in diverse environments in both saprophytic and parasitic phases.
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Paracoccidioides, Cell wall, Chitinase, Chromatography
Citação
SANTANA, Lidiane Aparecida da Penha; VAINSTEIN, Marilene Henning; TOMAZETT, Patrícia Kott; SANTOS-SILVA, Ludier Kesser; GÓES, Alfredo Miranda; SCHRANK, Augusto; SOARES, Célia Maria de Almeida; PEREIRA, Maristela. Distinct chitinases are expressed during various growth phases of the human pathogen Paracoccidioides brasiliensis. Memórias do Instituto Oswaldo Cruz, Rio de Janeiro, v. 107, n. 3, p. 310-316, May 2012.