Synthetic biology applied to Rhodosporidium toruloides for fine chemical production

• Autor(a) principal: Nora, Luísa Czamanski
• Data de Publicação: 2023
• Tipo de documento: Tese
• Idioma: eng
• Título da fonte: Biblioteca Digital de Teses e Dissertações da USP
• Texto Completo: https://www.teses.usp.br/teses/disponiveis/17/17136/tde-11042023-094354/
• topic: Biologia sintética; Cana-de-açúcar; Engenharia metabólica; Fatores de transcrição; Fungi; Fungos; Geraniol; Geraniol; Linalol; Linalool; Metabolic engineering; Monoterpene; Monoterpeno; Pinene; Pineno; Sugarcane; Synthetic biology; Transcription factors; Transcriptômica; Transcriptomics

The current increase in world population and standard of living is taking its toll on the planet\'s resources. The circular bioeconomy concept comes to spark the use of renewable biological resources to generate bio-based products, in order to reduce the use of fossil fuels and create a more sustainable way of living. In this context, microorganisms that can produce highly valuable chemicals using renewable feedstock are in high demand for applications in biorefineries. The combination of synthetic biology, bioinformatics and metabolic engineering strategies allows the optimization of hosts that can become microbial cell factories. The non-conventional yeast Rhodosporidium toruloides is one of the microorganisms with great potential to be applied for this purpose, since it is able to grow in a wide range of substrates and to withstand some of the stresses caused by bioprocesses. Thus, this work sought to understand transcriptional behaviors of R. toruloides when growing in stress-related conditions and with sugarcane as substrate, using RNA sequencing. A bioinformatic pipeline was then developed aiming the discovery of novel cis-regulatory elements from the yeast transcriptomic data, a tool that can be applied to other microbial hosts in the future. This work also ventured at producing valuable chemicals using R. toruloides as a host, applying metabolic engineering techniques and using state-of-the-art assembly methods. Three important terpenes were chosen for this production: pinene, linalool and geraniol. Although they were not detected, the Design, Build, Test, Learn cycle was successfully applied to reveal what can be improved in future endeavors. This work demonstrates methods for the interpretation of transcriptomic data, the detection of regulatory elements in nonconventional organisms, and the engineering of hosts to produce valuable chemicals. These strategies can be further optimized to create microbial cell factories for production of valuable chemicals in a green, renewable way, bringing our world to a more sustainable reality.