Directed Evolution of Membrane Transport Using Synthetic Selections.
ACS Synth Biol
; 7(3): 789-793, 2018 03 16.
Article
em En
| MEDLINE
| ID: mdl-29474058
ABSTRACT
Understanding and engineering solute transporters is important for metabolic engineering and the development of therapeutics. However, limited available experimental data on membrane transporters makes sequence-function relationships complex to predict. Here we apply ligand-responsive biosensor systems that enable selective growth of E. coli cells only if they functionally express an importer that is specific to the biosensor ligand. Using this system in a directed evolution framework, we successfully engineer the specificity of nicotinamide riboside transporters, PnuC, to accept thiamine as a substrate. Our results provide insight into the molecular determinants of substrate recognition of the PnuC transporter family and demonstrate how synthetic biology can be deployed to engineer the substrate spectrum of small molecule transporters.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Membrana Transportadoras
/
Evolução Molecular Direcionada
Idioma:
En
Revista:
ACS Synth Biol
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Dinamarca