Improvement of bioactive metabolite production in microbial cultures-A systems approach by OSMAC and deconvolution-based (HNMR)-H-1 quantification

Author(s)

Selegato, Denise Medeiros ^[1] ; Freire, Rafael Teixeira ^[2] ; Pilon, Alan Cesar ^[3] ; Biasetto, Carolina Rabal ^[1] ; de Oliveira, Haroldo Cesar ^[4] ; de Abreu, Lucas Magalhaes ^[5] ; Araujo, Angela Regina ^[1] ; Bolzani, Vanderlan da Silva ^[1] ; Castro-Gamboa, Ian ^[1]

Affiliation

^[1] Sao Paulo State Univ UNESP, Nucleus Bioassays Biosynth & Ecophysiol Nat Prod, Dept Organ Chem, Inst Chem, St Prof Francisco Degni 55, BR-14800060 Sao Paulo - Brazil

^[2] Univ Greenwich, Medway Metabon Res Grp, Chatham, Kent - England

^[3] Sao Paulo Univ USP, NPPNS, Fac Ciencias Farmaceut, Sao Paulo - Brazil

^[4] Sao Paulo State Univ UNESP, Lab Micol Clin, Nucleo Prote, Fac Ciencias Farmaceut Araraquara, Sao Paulo - Brazil

^[5] Univ Vicosa UFV, Dept Fisiopatol, Vicosa, MG - Brazil

• Document type: Journal article
• Source: Magnetic Resonance in Chemistry; v. 57, n. 8, p. 458-471, AUG 2019.
• Web of Science Citations: 0

Abstract

Traditionally, the screening of metabolites in microbial matrices is performed by monocultures. Nonetheless, the absence of biotic and abiotic interactions generally observed in nature still limit the chemical diversity and leads to ``poorer{''} chemical profiles. Nowadays, several methods have been developed to determine the conditions under which cryptic genes are activated, in an attempt to induce these silenced biosynthetic pathways. Among those, the one strain, many compounds (OSMAC) strategy has been applied to enhance metabolic production by a systematic variation of growth parameters. The complexity of the chemical profiles from OSMAC experiments has required increasingly robust and accurate techniques. In this sense, deconvolution-based (HNMR)-H-1 quantification have emerged as a promising methodology to decrease complexity and provide a comprehensive perspective for metabolomics studies. Our present work shows an integrated strategy for the increased production and rapid quantification of compounds from microbial sources. Specifically, an OSMAC design of experiments (DoE) was used to optimize the microbial production of bioactive fusaric acid, cytochalasin D and 3-nitropropionic acid, and Global Spectral Deconvolution (GSD)-based (HNMR)-H-1 quantification was carried out for their measurement. The results showed that OSMAC increased the production of the metabolites by up to 33% and that GSD was able to extract accurate NMR integrals even in heavily coalescence spectral regions. Moreover, GSD-(HNMR)-H-1 quantification was reproducible for all species and exhibited validated results that were more selective and accurate than comparative methods. Overall, this strategy up-regulated important metabolites using a reduced number of experiments and provided fast analyte monitor directly in raw extracts. (AU)

• FAPESP's process: 16/13292-8 - Analysis of nitrogen content in antropic influenced areas in Amazon and Atlantic Forest using a metabolomic tools
• Grantee: Alan Cesar Pilon
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 13/07600-3 - CIBFar - Center for Innovation in Biodiversity and Drug Discovery
• Grantee: Glaucius Oliva
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 15/14023-8 - Use of peptides with anti-adhesive activity in Paracoccidoides spp. in the treatment and prophylaxis of the paracoccidioidomycosis
• Grantee: Haroldo Cesar de Oliveira
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/05935-0 - Co-culture of microorganisms isolated from the rhizosphere of Senna spectabilis aiming to produce new bioactive metabolites
• Grantee: Denise Medeiros Selegato
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 09/54083-9 - Heteronuclear multidimensional NMR (16.4 Tesla): a new concept in the research in structural chemistry of natural and synthetic materials
• Grantee: Vanderlan da Silva Bolzani
• Support Opportunities: Multi-user Equipment Program