Estudo estrutural e molecular de exoglucanases de Moniliophthora perniciosa (Stahel) Aime & Phillips-Mora

Abstract

Sugarcane bagasse, a lignocellulosic residue generated in sugar-ethanol industries, is composed of polysaccharides that can be converted into glucose of industrial interest through the action of cellulases. Moniliophthora perniciosa, the causal agent of the plant disease “witch’s broom,” shows potential for synthesizing these biomolecules. This study aimed to characterize M. perniciosa exoglucanases through in silico and structural analyses, as well as to evaluate the expression of the genes encoding these enzymes, assessing their biotechnological potential. Accordingly, the study involved the identification and in silico characterization of exoglucanases, whose three- dimensional structures were predicted by comparative modeling. Subsequently, molecular analyses were performed. Specific primers were initially designed for the genes encoding the proteins under investigation. Next, high-quality RNA extracted from the medium containing lignocellulosic material was used for cDNA synthesis, which was later amplified, and the resulting PCR product was purified. The analyzed sequences were identified as belonging to the GH6 family, displaying conserved regions typical of the group. The generated three-dimensional models showed high structural quality, with preservation of essential functional regions, highlighting the biotechnological potential of these enzymes. The fungal cultivation conditions in lignocellulosic medium proved favorable for the induction of cellulase expression, and expression of the gene corresponding to the exoglucanase was detected. These results indicate that the cDNA sequence obtained constitutes a promising candidate for heterologous expression, purification, and characterization, reinforcing the potential of Moniliophthora perniciosa cellulases in optimizing sustainable industrial bioprocesses.