Identificação de uma nova fitase bacteriana através de prospecção metagenômica

Abstract

The bacterial diversity of environmental samples and their potencial are still poorly explored. Metagenomics approaches offer the opportunity for the prospection of genes with biotechnological interest from non-cultured microorganisms in the laboratory. Materials in the process of composting exhibit high microbiological activity, which indicates that the environment is conducive to the degradation of lignocellulosic material. In this context, several enzymes have been identified by catalyzing these reactions. Phytases are widely produced phosphatase enzymes that hydrolyze phytate by releasing phosphorus and chelated minerals. Due to this potential, these enzymes have become easily interesting for the biotech field. The aim was to investigate the metagenomic potential for the identification of genes with phytase activity. After obtaining the material, DNA extraction was performed, followed by its purification. Total extracted DNA was used to construct the metagenomic library with cosmid vector through commercial kits (EPICENTRE). Positive clones for phytase activity were sequenced and analyzed. The prospection carried out in this study identified a gene (PhyRC001) encoding a phytase where the amino acid identity between PhyRC001 and its nearest published homologs is less than 60%.