| Congresso Brasileiro de Microbiologia 2023 | Resumo: 484-1 | ||||
Resumo:Cyanobacteria is a very ancient prokaryotic phylum, which has a wide morphological diversity and ecological importance. Pantanalinema is a genus of cyanobacteria belonging to the Leptolynbyaceae family, which was originally described in saline alkaline waters of the Pantanal, biome. Recently, one isolate of the genus was described in the Cerrado biome, isolated from a waterfall on the outer edge of Chapada das Mesas National Park, located in the south of Maranhão, being the first sequenced genome of the group and available under accession number GCA_016743235.1. This work aims to characterize hypothetical proteins presents in the biosynthetic metabolic pathways of the genome of the cyanobacterium Pantanalinema sp. GBBB05. The draft genome was submitted to antiSMASH v.6.0, with extra features “All on”. A total of 14 gene clusters were identified with 80 hypothetical proteins (HPs) mapped. The HPs were manually curated seeking to identify domains and superfamilies through the tools BLASTp, CDD, Pfam, SMART, CATH, UniProt and InterPro Moving on to the analysis of physical-chemical parameters by Expasy's ProtParam server; the subcellular location measured by PSORTb v.3.0.3, PSLPred and CELLO, the SecretomeP 2.0 and SinalP 6.0 servers for predicting secretory proteins and signal peptides; the TMHMM and CCTOP servers were used to predict transmembrane regions and topology, and SOSUI characterized the proteins as membrane or soluble. To assess the reliability of the forecast, the ROC curve was build, through the Online ROC CURVE Calculator. In addition, gene trees were built through MEGA Version X, and synteny analysis, with Clinker and Clustermap.js, to understand the evolutionary relationship with other organisms. Among the gene clusters identified by AntiSMASH 6.0. are terpenes, beta-lactone, resorcinol and NRPS/NRPS-like/RiPP pathways. Eighty hypothetical proteins (HPs) were obtained, 16 of which were classified as High Confidence Proteins (HCs), with domains from the NnrU family, OprB, Cysteine peptidase, bacteriocins, phycobilisomes, peroxiredoxin and AhpC/TSA.
The analyzes of physical-chemical parameters indicated that 50% of the HCs are stable and most of them hydropathic; the subcellular location of the proteins are mostly in the cytoplasmic domain (60%); only four proteins had secretory activity and two with signal peptide; and 25% were identified as transmembrane proteins. The evaluation of the performance of the databases indicated that the CDD was the predictor with the highest mean sensitivity, specificity and accuracy. Phylogenomic analysis was performed with 19 other strains, confirming proximity to Pantanalinema sp. GBBB05 as a member of the Leptolyngbtaceae family, close to two species of the genus Leptolyngbya. Synteny analysis indicated that GBBB05 shares PKS_KS domain proteins with 36% identity and heterocyclization domain with 44%, with the Fischerella sp. PCC 9431 strain. Through the results obtained, it is possible to take initial steps towards a deeper genomic characterization of Pantanalinema sp. GBBB05, contributing to the characterization of local diversity and is valorization through biotechnological potential. Palavras-chave: Biosynthetic metabolic pathways, Gene Clusters, Genome mining, ROC curve, Synteny Agência de fomento:Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). | |||||