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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
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The directed evolution of NDM-1
Caitlyn A Thomas1, Zishuo Cheng1, Christopher R Bethel2
1Department of Chemistry and Biochemistry, Miami University , Oxford, Ohio, USA.
Antimicrobial Agents and Chemotherapy
|October 24, 2023
Summary
New Delhi Metallo-β-lactamase (NDM-1) variants did not increase resistance to meropenem via amino acid changes. Increased transcription or altered zinc transport may be more critical for meropenem resistance in clinical settings.
Area of Science:
- Microbiology
- Enzymology
- Drug Resistance
Background:
- β-Lactam antibiotics are crucial therapeutics, but resistance is a growing concern.
- β-Lactamases, particularly metallo-β-lactamases (MBLs), are key enzymes conferring resistance by hydrolyzing β-lactam bonds.
- New Delhi Metallo-β-lactamase (NDM-1) is a clinically significant MBL requiring further study.
Purpose of the Study:
- To investigate the in vitro protein evolution of NDM-1 β-lactamase.
- To identify if amino acid substitutions in NDM-1 can enhance resistance to meropenem.
Main Methods:
- Utilized error-prone polymerase chain reaction to generate NDM-1 variants.
- Evaluated the resistance profiles of generated variants against meropenem.
Main Results:
- No observed amino acid substitutions in NDM-1 variants conferred increased resistance to meropenem.
- Protein evolution through amino acid changes does not appear to be a primary driver of meropenem resistance for NDM-1.
Conclusions:
- Amino acid substitutions in NDM-1 are unlikely to be the main mechanism for increased meropenem resistance.
- Alternative mechanisms, such as increased gene transcription or altered zinc ion transport, warrant further investigation for clinical relevance in NDM-1 mediated resistance.
