| Congresso Brasileiro de Microbiologia 2023 | Resumo: 99-1 | ||||
Resumo:Polyurethanes (PUs) are plastic polymers that represent an environmental pollution problem. The search for microorganisms or enzymes to carry out the biological recycling of these polymers is on the rise. The isolation of microorganisms from the gut of larvae of different animal species for plastic biodegradation is an attractive strategy. Thus, this work aimed to isolate and identify a bacteria with the potential for biodegradation of PUs. For this, the gut of Galleria mellonella larvae was inoculated in Luria Bertani (LB) broth supplemented with 0.1, 1, and 5% (w/v) of Impranil® PU and incubated for seven days at 30 °C. Subsequently, serial dilutions were plated on LB agar supplemented with 1% Impranil®. The colony that showed a transparent halo around it was submitted for purification and identification. Genomic DNA was extracted using the Promega Wizard® DNA extraction kit. DNA quality and quantity were evaluated by 0.8% agarose gel electrophoresis and Qubit™ 2.0 fluorometer, respectively. Genome sequencing was performed by the commercial company Novogene (California, USA) using the Pacbio Sequel platform (Illumina PE150). The quality control of the sequencing data, assembly, and genome annotation was conducted. The Rapid Annotation Subsystem Technology (RAST) server classified the predicted genes into the subsystems of the SEED database. For a taxonomic assignment, the JspeciesWS server compared the S. warneri genome with reference genomes and calculated two Average Nucleotide Identity indexes based on BLAST (ANIb) and MUMMER (ANIm), using default parameters. The Genome-to-Genome Distance Calculator (GGDC) server version 3.0 calculated the digital DNA-DNA hybridization (dDDH) using default parameters. The bacterium isolated from the gut of the larvae of G. mellonella was identified, based on the sequencing of the complete genome, as belonging to the species Staphylococcus warneri. The isolate was named S. warneri UFV_01.21, and the BioProject and BioSample accession numbers are PRJNA875469 and SAMN30619083, respectively. The genome length of S. warneri UFV_01.21 was determined to be 2,544,871 bp, distributed in 25 contigs (22 chromosomal and three plasmidial) and a GC average content of 32.7%. In the genome assembly, 2,438 Coding DNA Sequences (CDS), 33 tRNA genes, nine rRNA genes, and one tmRNA gene were predicted. With the RAST server, it was possible to predict 1,318 CDS (54.06%) according to the functional category, the most abundant being: cofactors, vitamins, prosthetic groups, pigments (192 genes), protein metabolism (163 genes), amino acids and derivatives (122 genes), virulence, disease and defense (88 genes), carbohydrates (86 genes), RNA metabolism (86 genes) and DNA metabolism (75 genes). Comparative genomic analysis of the S. warneri UFV_01.21 genome with other S. warneri genomes guided its taxonomic classification, which was higher than the threshold score of 95~96% ANI and the cutoff value of 70% dDDH. This is the first study that describes the isolation and draft genome of S. warneri from the gut of the larvae of G. melonella, which can degrade Impranil® PU, emphasizing that the potential for biodegradation of PUs by this isolate must be explored. Palavras-chave: bacterial genome, gut microbiota of larvas, biodegradation Agência de fomento:CNPq, CAPES, FAPEMIG | |||||