Análise das proteínas glicosiladas secretadas pelo isolado trichoderma harzianum (ALL-42) induzida por parede celular de fitopatógenos

Abstract

Glycoproteins are molecules formed by the union of proteins to glycans by a post-translational process called glycosylation. This process is carried out in a variety of cell types, including fungal cell and the produced glycoproteins have biological roles essential for life. Trichoderma harzianum (ALL-42) is a fungus widely studied due to its role of biocontrol and plant growth induction. Studies involving substances produced by this microorganism may contribute to the understanding of this mechanism. The aim of this study was to identify and analyze the glycosylated proteins secreted by T. harzianum when grown in culture medium containing cell walls of different pathogen. The secretomas produced for walls of Sclerotinia sclerotiorum and Fusarium oxysporum were then subjected to Concanavalin A affinity chromatography and subsequently carried to SDS-PAGE electrophoresis and staining with Pro-Q Emerald 488 for identifying glycoproteins. The bands displayed in the gel were cut out and subjected to mass spectrometry. The results showed the presence of some proteins, galactose oxysdases, glycoside hydrolases, β-1,3-exoglucanases e proteases, with important functions in enzymatic cell wall degradation mechanism of pathogen and the presence of these proteins coincides with results from other studies. The samples showed enzymatic activity for seven substrates tested, chosen in accordance with the enzymes identified by mass spectrometry and analyzing the relative gene expression. Genes related to these proteins were identified in glicosecretoma found in higher expression as T. harzianum (ALL -42 ) was grown on the walls of plant pathogens compared to two other situations , grown in glucose and glycerol as carbon sources.