研究目的
To develop a fluorescence assay for the rapid screening of metallo-β-lactamase (MBL) inhibitors using a fluorogenic carbapenem CPC-1 as substrate, compatible with all MBLs, including B1, B2 and B3 subclass MBLs.
研究成果
The CPC-1-based fluorescence assay is a sensitive and general method for screening MBL inhibitors, compatible with all MBL subclasses. 2,3-dimercaprol was identified as a potent inhibitor for CphA, suggesting its potential as a broad-spectrum MBL inhibitor.
研究不足
The study focuses on in vitro screening and does not address in vivo efficacy or toxicity of identified inhibitors.
1：Experimental Design and Method Selection:
The study employs a fluorescence-based assay using CPC-1 as a substrate for MBL activity detection.
2：Sample Selection and Data Sources:
Recombinant MBLs (NDM-1, IMP-1, CphA, L1) were produced in E. coli.
3：List of Experimental Equipment and Materials:
Fluorogenic carbapenem CPC-1, recombinant MBLs, various inhibitors.
4：Experimental Procedures and Operational Workflow:
Fluorescence intensity measurements were conducted to monitor CPC-1 hydrolysis by MBLs in the presence of potential inhibitors.
5：Data Analysis Methods:
Kinetic parameters (Km, kcat) and inhibitory concentrations (IC50) were determined using fluorescence data.
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