| Texto completo | |
| Autor(es): |
Chaves, Gabriel Luz
;
Batista, Raquel Salgado
;
Cunha, Josivan de Sousa
;
Oliveira, Davi Benedito
;
da Silva, Mateus Ribeiro
;
Deriggi Pisani, Graziele Fernanda
;
Selistre-de-Araujo, Heloisa Sobreiro
;
Zangirolami, Teresa Cristina
;
da Silva, Adilson Jose
Número total de Autores: 9
|
| Tipo de documento: | Artigo Científico |
| Fonte: | NEW BIOTECHNOLOGY; v. 72, p. 9-pg., 2022-10-25. |
| Resumo | |
3-Hydroxypropionic acid (3-HP) production from renewable feedstocks is of great interest in efforts to develop greener processes for obtaining this chemical platform. Here we report an engineered E. coli strain for 3-HP production through the beta-alanine pathway. To obtain a new strain capable of producing 3-HP, the pathway was established by overexpressing the enzymes pyruvate aminotransferase, 3-hydroxyacid dehydrogenase, and L-aspartate-1-decarboxylase. Further increase of the 3-HP titer was achieved using evolutionary optimizations of a genome-scale metabolic model of E. coli containing the adopted pathway. From these optimizations, three non -intuitive targets for in vivo assessment were identified: L-alanine aminotransferase and alanine racemase over -expression, and L-valine transaminase knock-out. The implementation of these targets in the production strain resulted in a 40% increase in 3-HP titer. The strain was further engineered to overexpress phosphoenolpyruvate carboxylase, reaching 0.79 +/- 0.02 g/L of 3-HP when grown using glucose. Surprisingly, this strain produced 63% more of the desired product when grown using a mixture of glucose and xylose (1:1, C-mol), and gene expression analysis showed that the cellular adjustment to consume xylose had a positive impact on 3-HP accumulation. Fed-batch culture with xylose feeding led to a final titer of 29.1 g/L. These results reinforce the value of computational methods in strain engineering, enabling the design of more efficient strategies to be assessed. Moreover, higher production of 3-HP under a sugar mixture condition points towards the development of bio-processes based on renewable resources, such as hemicellulose hydrolysates. (AU) | |
| Processo FAPESP: | 16/10636-8 - Da fábrica celular à biorrefinaria integrada Biodiesel-Bioetanol: uma abordagem sistêmica aplicada a problemas complexos em micro e macroescalas |
| Beneficiário: | Roberto de Campos Giordano |
| Modalidade de apoio: | Auxílio à Pesquisa - Programa BIOEN - Temático |
| Processo FAPESP: | 19/07902-6 - Construção de uma fábrica celular para produção de ácido 3-hidroxipropiônico |
| Beneficiário: | Gabriel Luz Chaves |
| Modalidade de apoio: | Bolsas no Brasil - Mestrado |