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Published on: August 18, 2023
Directed evolution of trimethoprim resistance in Escherichia coli
Morgan Watson1, Jian-Wei Liu, David Ollis
1Research School of Chemistry, Australian National University, Canberra, ACT, Australia. watson@rsc.anu.edu.au
Abstract:
Directed evolution is a useful tool in the study of enzymes. It is used in this study to investigate the means by which resistance to the antibiotic trimethoprim develops in dihyrofolate reductase from Escherichia coli. Mutants with clinical levels of resistance were obtained after only three generations. After four generations of directed evolution, several mutants were characterized, along with some point mutants made to investigate amino acid changes of interest. Several mutations were found to grant resistance to trimethoprim, both by reducing the binding affinity of the enzyme for the drug, and by increasing the activity of the enzyme.
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