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Sequential mass spectrometry applied to the monitoring of induction of bioactive molecules in microbial co-culture

Grant number: 18/03782-3
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): August 01, 2018
Effective date (End): July 31, 2022
Field of knowledge:Physical Sciences and Mathematics - Chemistry
Principal Investigator:Luiz Alberto Beraldo de Moraes
Grantee:Talita Carla de Tralia Medeiros
Home Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Actinobacteria, especially of the genre Streptomyces, are an inexhaustible source of metabolites bioactive secondary substances, particularly for the discovery of new active principles of interest industrial. Although actinobacteria have the ability to biosynthesize a large variety of molecules, recent advances in genome sequencing have revealed that these micro-organisms have an even greater potential for producing secondary metabolites with a still unknown. Thus, many microbial genes can remain silenced "cryptic genes" under conditions of artificial growth in the laboratory. We the last ten years, several methods have been developed to aid in the activation of this biosynthesis. As an alternative, several strategies have been applied for the activation mutated microbial pathways, such as OSMAC, heterologous and homologous expression, ribosomal and co-culture. Co-cultivation is based on the cultivation of two or more microorganisms confined in the same environment, mimicking the microbial communities present in nature. Where these communities interact with each other through the release of signaling molecules or defense. These interactions may induce the activation of silenced pathways in isolated culture. Like this, these molecules, produced dynamically, are of great interest for the discovery of bioactive molecules. Therefore, this doctoral project has as main objective to explore the analytical versatility of mass spectrometry in its most varied scanning modes, for the monitoring of the secondary metabolites produced by the activation of silenced pathways different activation strategies, such as co-cultivation, mixed cultivation, addition of inducers. Mass Spectrometry (MS) is the most specific technique for the detection and identification of secondary metabolites. MS is able to provide mass information accurate molecular structure, as well as a wealth of structural details that together provide a unique fingerprint for each constituent of the sample, accelerating the identification of molecules present in complex matrices. (AU)