Characterization of wcb cluster from b. seminalis TC3.4.2R3 and its role in the synthesis of antimicrobial compounds

• Grant number: 14/24375-6
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: April 01, 2015
• End date: September 30, 2015
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Welington Luiz de Araújo
• Grantee: Priscila Jane Romano de Oliveira Gonçalves
• Supervisor: Brendan Wren
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: London School of Hygiene and Tropical Medicine, England
• Associated to the scholarship: 13/03426-9 - Characterization of wcb cluster of B. seminalis strains TC3.4.2R3 and its role in the biosynthesis of antimicrobial compounds, BP.DR

Abstract

Several compounds with antimicrobial activity produced by Burkholderia spp. have been investigated, which could replace conventional chemical pesticides. Gene clusters of Burkholderia seminalis strain TC3.4.2R3 associated to the synthesis of antimicrobial compounds have been identified by analyzing a mutant library. Among these genes we identified a gene encoding a glycosyltransferase (wcbE) in the wcb cluster, which is associated with capsule synthesis and microbial interactions. Comparison of the wcb cluster region with other species of Burkholderia genus showed that some genes are not shared with other Burkholderia species, such as a gene encoding a hypothetical protein and epimerase gene. Another gene, named wcbR is conserved in this genus and has a PKS domain, suggesting that this cluster may be associated not only with the synthesis of capsule, but also with the synthesis of secondary metabolites. Phenotypic tests revealed that mutants of the wcbE glycosyltransferase lose the ability to inhibit pathogens compared to wild-type B. seminalis. The antagonism test with organic extracts from B. seminalis both wild-type and some mutants showed inhibition of fingal growth, suggesting that molecules with antimicrobial activity are extracted in the process. Thus, studies with B. seminalis can contribute to understanding the relationship between the synthesis of secondary metabolites and wcbE gene, as well as others genes from wcb cluster. Thereby, the research internship abroad aims to obtain and analyze mutants with deleted glycosyltransferase gene and other genes selected from wcb cluster, such as a hypothetical protein, the epimerase gene and the wcbR gene. Furthermore, mutant and wild strains will be assessed for biofilm formation, motility and oxidative stress, which will allow the understanding of the mechanisms related to the interaction between B. seminalis TC3.4.2R3 and other microorganisms in the environment. The research will be conducted in the Department of Infectious & Tropical Diseases, London School of Hygiene & Tropical Medicine under supervision of Prof. Dr. Brendan Wren, who has extensive experience in the study of the bacterial glycobiology at the molecular level. The results will contribute to the enrichment of the candidate thesis, which follows with chemistry and metabolomics analysis of extracts with antimicrobial activity obtained from B. seminalis. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ROMANO DE OLIVEIRA GONCALVES, PRISCILA JANE; HUME, CARMEN C. DENMAN; FERREIRA, ALMIR JOSE; TSUI, SARINA; BROCCHI, MARCELO; WREN, BRENDAN W.; ARAUJO, WELINGTON LUIZ. Environmental interactions are regulated by temperature in Burkholderia seminalis TC3.4.2R3. SCIENTIFIC REPORTS, v. 9, APR 2 2019. (17/50447-2, 12/24217-6, 13/03426-9, 14/24375-6, 17/12510-4)