Engineering a therapeutic microbe for site-of-infection delivery of encapsulated antimicrobial peptides (AMPs)
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
Antimicrobial peptides (AMPs) are promising therapeutics against antibiotic resistant pathogens, but they are difficult to administer at the site of infection due to poor stability. To enhance the stability of AMPs for effective and specific delivery at a site of infection, this project aimed to develop components of a novel, cell-based, drug delivery platform for AMPs in which a therapeutic microbe is engineered to specifically detect a pathogen of interest and effectively release AMPs at high, local concentrations in response. We focused the majority of our project on developing a system to produce normally host-toxic AMPs in our model therapeutic microbe (Escherichia coli) in high quantities using a protein cage system called an encapsulin (Enc) nanocompartment system. We were able to successfully produce an AMP called HBCM2 (HB), which is effective in killing our model pathogen, Pseudomonas aeruginosa, in E. coli by fusing it to an engineered, non-cage forming Enc protein, which prevented its toxicity to the E. coli. This engineered Enc protein could then be digested by a specific protease (TEV protease) for release of HB when desired. In addition to the HB production system, we also constructed a preliminary engineered E. coli capable of sensing P. aeruginosa via its specific quorum-sensing molecule, producing HB peptide, and releasing it via self-lysis in response. Preliminary characterization of this system was conducted, but we hope to fully characterize the system in future studies. The overall results produced from this study demonstrate the power of synthetic biology to confer novel functions on cells for biomanufacturing and therapeutic applications.
- Research Organization:
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
- Sponsoring Organization:
- USDOE National Nuclear Security Administration (NNSA)
- DOE Contract Number:
- AC52-07NA27344
- OSTI ID:
- 1573149
- Report Number(s):
- LLNL-TR-795839; 997177
- Country of Publication:
- United States
- Language:
- English
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