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| Author(s): |
Douglas Cunha Sachito
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Instituto de Química |
| Defense date: | 2024-10-02 |
| Examining board members: |
Ljubica Tasic;
Katlin Brauer Massirer;
Elizabeth Bilsland;
Camila Manoel Crnkovic;
Mônica Tallarico Pupo
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| Advisor: | Ljubica Tasic; Luciana Gonzaga de Oliveira |
| Abstract | |
Terpenes are secondary metabolites with wide-ranging industrial applications and represent the most abundant class of natural products (NPs) in nature. However, bacterial terpenes have been largely overlooked for decades despite their significant biotechnological potential. The advent of next-generation sequencing (NGS) technologies and the reduced costs of genome sequencing have made it possible to explore the vast potential of bacterial genomes for terpene biosynthesis. This study aimed to analyze bacterial terpenes produced by an actinomycete isolated from Brazilian territory. Specifically, the research focused on genome mining of Streptomyces sp. CBMAI 2042, an endophyte of Citrus sinensis. This analysis revealed 37 gene clusters associated with secondary metabolite production, including five terpene clusters: three for sesquiterpenes, one for hopanoids, and one for carotenoids. The transcription of target genes and the identification of 28 volatile sesquiterpenes in extracts from the mycelium of Streptomyces sp. CBMAI 2042 were investigated. Additionally, gene-metabolite associations were established using genetic disruption engineering. The research also included cloning, production, and characterization of recombinant terpene cyclases (TCs), enzTS-1-4, from Streptomyces sp. CBMAI 2042. Ideal reaction conditions and the effects of ammonium hydroxide on modulating terpene production by enzTS-2 and enzTS-3 were evaluated. Notably, enzTS-2 and enzTS-3 demonstrated versatility by accepting analogs of farnesyl pyrophosphate as well as C10 substrates without requiring enzymatic evolution. This versatility extends the applications of these bacterial TCs, suggesting their potential for optimizing the production of analogs with innovative applications. These findings underscore the importance of studying Brazilian soil microorganisms, such as Streptomyces, for discovering new enzymes and bioactive molecules. The genome mining strategy proved to be a powerful, fast, and effective approach for identifying terpene clusters in bacteria. Future research could expand the biotechnological applications of sesquiterpenes, their analogs, and the TCs investigated in this study, contributing to societal advancements. This work highlights the untapped potential of bacterial terpenes and demonstrates Brazil's capacity, with its vast biodiversity, to become a global leader in biotechnological innovation, fostering the development of sustainable products and technologies (AU) | |
| FAPESP's process: | 19/04855-7 - Exploration and expanding the terpenome of Streptomyces sp. CBMAI 2042 via genome mining |
| Grantee: | Douglas Cunha Sachito |
| Support Opportunities: | Scholarships in Brazil - Doctorate (Direct) |