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Genetic Toolbox Expansion Enables Constitutively Fluorescent Lacticaseibacillus rhamnosus for Functional Microbiome
Ilke Van Tente1, Marc Blanch-Asensio2,3, Tom Eilers1
1Laboratory of Applied Microbiology and Biotechnology, Department of Bioscience Engineering, University of Antwerp, Antwerp, Belgium.
Microbial Biotechnology
|July 10, 2026
Summary
Researchers enhanced the genetic tools for Lacticaseibacillus rhamnosus probiotics, enabling new applications in synthetic biology and microbiome research. This work provides valuable resources for studying probiotic functions and interactions.
Area of Science:
- Microbiology
- Synthetic Biology
- Genetics
Background:
- Lacticaseibacillus rhamnosus strains are recognized probiotics with limited genetic tools, hindering research.
- Existing genetic tools are insufficient for advanced functional microbiome studies and synthetic biology.
Purpose of the Study:
- To expand the genetic toolbox for L. rhamnosus GR-1 and L. rhamnosus GG.
- To implement and validate genetic elements from Lactiplantibacillus plantarum.
- To engineer fluorescent L. rhamnosus strains for functional studies.
Main Methods:
- Direct plasmid cloning and testing of genetic elements.
- Evaluation of five constitutive promoters (PtlpA, Ptec, PcpG, P48, P23).
- Characterization of PtlpA promoter activity, thermo-responsiveness, and stability.
- Engineering of L. rhamnosus GR-1 with fluorescent proteins (mCherry, mScarlet3, sfGFP).
Main Results:
- PtlpA demonstrated strong constitutive promoter activity in L. rhamnosus GR-1.
- The PtlpA promoter's functionality was enhanced with a repressor and characterized for industrial applications.
- Successfully engineered constitutively fluorescent L. rhamnosus GR-1 strains.
- Demonstrated functionality of fluorescent strains in a competitive growth assay against Staphylococcus aureus.
Conclusions:
- The expanded genetic toolkit and fluorescent L. rhamnosus strains are valuable resources for microbiome research.
- These tools facilitate the study of probiotic functional properties like adhesion and host-microbe interactions.
- Advances Lactobacillaceae as a chassis for synthetic biology applications.
