Directed Evolution of Membrane Transport Using Synthetic Selections
Autor: | Hans Jasper Genee, Anne Pihl Bali, Morten Otto Alexander Sommer |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
030102 biochemistry & molecular biology Biomedical Engineering Membrane Transport Proteins Transporter General Medicine Computational biology Membrane transport Directed evolution Biochemistry Genetics and Molecular Biology (miscellaneous) Small molecule Substrate Specificity Metabolic engineering 03 medical and health sciences chemistry.chemical_compound Synthetic biology 030104 developmental biology Structural biology chemistry Nicotinamide riboside Escherichia coli Amino Acid Sequence Thiamine Directed Molecular Evolution |
Zdroj: | ACS Synthetic Biology. 7:789-793 |
ISSN: | 2161-5063 |
Popis: | 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. |
Databáze: | OpenAIRE |
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