Investigação da produção de lipases por um consórcio bacteriano isolado da biodiversidade amazônica

Abstract

Microorganisms produce a variety of bioproducts, such as enzymes, which function as biocatalysts—highly specific molecules that are less polluting when compared to chemical catalysts and operate through lower-energy activation pathways, thereby accelerating chemical reactions. These microorganisms can be naturally found in soil or aquatic environments and can also be cultivated under laboratory conditions to evaluate their enzymatic activity potential. Lipase is an enzyme that hydrolyzes fatty acid esters, including triglycerides, the primary components of oil resulting in the formation of free fatty acids and glycerol. These hydrolysates are simpler compounds and less harmful to the environment. Considering that waste frying oil constitutes a significant environmental pollutant, this dissertation aimed to select bacteria isolated from Amazonian biodiversity capable of utilizing waste frying oil as a carbon source, which, in future approaches, may be applied in bioremediation strategies. Initially, based on the genome sequences of these bacteria, the presence of lipase-coding genes was investigated. Genomic annotation performed using the RAST tool enabled the identification of potential genes related to lipolytic degradation, metabolic pathways through KEGG via the KBase platform, and antibiotic resistance genes via CARD, with only one resistance result detected in Pseudomonas sp. Lipase production from waste frying oil was evaluated through enzymatic activity assays using pNPP as substrate and by protein profiling via SDS-PAGE. The highest lipolytic activities were observed in Pseudomonas sp. and the bacterial consortium, with activities of 226 U/L and 259 U/L, respectively. Protein separation further allowed detection of bands within the expected molecular weight range in Pseudomonas sp. and A. seifertii.