Prospecção de enzimas a partir de consórcio microbiano degradador de petróleo

Abstract

Proteomics is considered an emerging biotechnological tool that helps to clarify the mechanisms involved in the biotransformation/biodegradation of hydrocarbons. Therefore, this work aimed to determine the differential enzymatic patterns of the microbial consortium degrading oil from the Recôncavo Baiano basin, in addition, this work is relevant for future systems in recovery of oil-impacted environments. The mixed microbial consortium was composed of 10 bacteria and 23 fungi of marine origin, which were identified by the Sanger method using the 16S and ITS genes. The consortium was subjected to constant agitation at 180 rpm at 35 °C for 7 days to assess crude oil depletion and identify enzymatic patterns in the presence of oil. The concentration of hydrocarbons was detected by gas chromatography coupled to a flame ionization detector according to the protocol adapted from USEPA 3510 and the degradation was evaluated by the ratios Pr/Ph, Pr/C17, Ph/C18, HTP/UCM and others. The peptides were sequenced using MS-MS liquid chromatography and the enzymes were identified using the PatternLab for Proteomics software. From the preliminary results, a reduction in resolved hydrocarbons and unresolved complex mixture hydrocarbons, as well as isoprenoids, pristane and phytane, was observed and could be verified. The proteomic analysis revealed the predominance of enzymes related to the stress response, transport and translation/transcription when in the presence of oil and indicated the presence of enzymes associated with the degradation of xenobiotics, indicating their possible use in biotechnological applications. Thus, constructing a library of proteins related to crude oil clarifies the degradation potential of the microbial consortium and lays the foundations for the optimizing of a more robust system of environmental recovery.