Figure 1. Expression and purification of soluble recombinant β-lactamase using Escherichia coli as the expression host and pET-28a as the cloning vector. (Li, Lele, et al. 2022)
Low doses of piperacillin treat murine Lyme disease without affecting the microbiome.
A high-resolution screen identifies a preexisting beta-lactam that specifically treats Lyme disease in mice
Paper
2025
Science Translational Medicine |
DOI
Background
Lyme disease, caused by Borrelia burgdorferi in the United States, is a growing public health concern. Doxycycline, the standard treatment, has limitations including side effects, restrictions, and treatment failures. The authors screened nearly 500 FDA-approved compounds using fluorescent D-alanine and cellular assays, and identified piperacillin, a beta-lactam antibiotic with potent activity against B. burgdorferi. Piperacillin killed spirochetes at nanomolar concentrations by interfering with their specific multizonal peptidoglycan synthesis and binding to the divisome protein BB0136. In mice, piperacillin effectively treated Lyme disease at a 100-fold lower dose than doxycycline without affecting the gut microbiome, offering an attractive alternative therapy.
Key Findings
The investigators established a high-resolution screening workflow using the fluorescent D-alanine analog HADA to monitor peptidoglycan synthesis in B. burgdorferi and evaluated 466 FDA-approved compounds. Piperacillin emerged as a highly selective candidate with an MIC of approximately 35 nM and superior killing compared with doxycycline in vitro. Single-cell imaging revealed that piperacillin abolished the characteristic zonal peptidoglycan synthesis and elongated cells, while FtsA-marker studies indicated that the target acts after initial divisome assembly. Competition assays identified BB0136 (putative FtsI/PBP3) as the molecular target, with an IC50 of 20.5 nM, and structural modeling suggested similarity to Streptococcus pneumoniae Pbp2X. In infected mice, piperacillin cleared infection at 1 mg/kg per day, a 100-fold lower dose than doxycycline, and no viable spirochetes recurred up to 8 weeks after treatment. Importantly, piperacillin did not significantly alter the fecal microbiome, unlike doxycycline or ceftriaxone. Overall, these data provide strong evidence that piperacillin is a specific, preexisting antibiotic that could be repurposed for Lyme disease.
Contribution
Creative Biogene contributed the pET28a expression vector used in this study to clone, express, and purify recombinant BB0136 in E. coli. This reagent was essential for Bocillin-FL competition binding assays and structural modeling, enabling the authors to establish piperacillin's molecular target and its selective anti-Borrelia mechanism.
Lyme disease, caused by Borrelia burgdorferi in the United States, is a growing public health concern. Doxycycline, the standard treatment, has limitations including side effects, restrictions, and treatment failures. The authors screened nearly 500 FDA-approved compounds using fluorescent D-alanine and cellular assays, and identified piperacillin, a beta-lactam antibiotic with potent activity against B. burgdorferi. Piperacillin killed spirochetes at nanomolar concentrations by interfering with their specific multizonal peptidoglycan synthesis and binding to the divisome protein BB0136. In mice, piperacillin effectively treated Lyme disease at a 100-fold lower dose than doxycycline without affecting the gut microbiome, offering an attractive alternative therapy.
Key Findings
The investigators established a high-resolution screening workflow using the fluorescent D-alanine analog HADA to monitor peptidoglycan synthesis in B. burgdorferi and evaluated 466 FDA-approved compounds. Piperacillin emerged as a highly selective candidate with an MIC of approximately 35 nM and superior killing compared with doxycycline in vitro. Single-cell imaging revealed that piperacillin abolished the characteristic zonal peptidoglycan synthesis and elongated cells, while FtsA-marker studies indicated that the target acts after initial divisome assembly. Competition assays identified BB0136 (putative FtsI/PBP3) as the molecular target, with an IC50 of 20.5 nM, and structural modeling suggested similarity to Streptococcus pneumoniae Pbp2X. In infected mice, piperacillin cleared infection at 1 mg/kg per day, a 100-fold lower dose than doxycycline, and no viable spirochetes recurred up to 8 weeks after treatment. Importantly, piperacillin did not significantly alter the fecal microbiome, unlike doxycycline or ceftriaxone. Overall, these data provide strong evidence that piperacillin is a specific, preexisting antibiotic that could be repurposed for Lyme disease.
Contribution
Creative Biogene contributed the pET28a expression vector used in this study to clone, express, and purify recombinant BB0136 in E. coli. This reagent was essential for Bocillin-FL competition binding assays and structural modeling, enabling the authors to establish piperacillin's molecular target and its selective anti-Borrelia mechanism.