RT Journal Article
SR Electronic
T1 Genetic and materials engineering to enhance inducible gene expression in lactobacilli
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2025.01.10.632477
DO 10.1101/2025.01.10.632477
A1 Blanch-Asensio, Marc
A1 Tadimarri, Varun Sai
A1 Martinez, Roberto
A1 Dahiya, Gurvinder Singh
A1 Lale, Rahmi
A1 Sankaran, Shrikrishnan
YR 2025
UL http://biorxiv.org/content/early/2025/01/13/2025.01.10.632477.abstract
AB Lactiplantibacillus plantarum, a versatile member of the lactobacilli family, is widely recognized for its potential in healthcare, food production, and environmental biotechnology. Despite its inherent advantages and demonstrated potential as microbial chassis for biotechnological applications, its broader utility is constrained by a limited genetic toolbox, particularly the lack of robust inducible gene expression systems. Addressing this gap, we report the development of a novel inducible system for L. plantarum based on a strong bacteriophage-derived promoter and the food-grade inducer, cumate. This system demonstrates enhanced dynamic range and temporal control of gene expression while exhibiting interesting temperature and growth phase dependent phenomena that influence leakiness and induced expression levels. Interestingly, encapsulation of engineered L. plantarum strains in alginate-based hydrogels significantly reduces leaky expression and maintains induction efficacy. Our findings highlight the synergistic potential of combining genetic engineering with encapsulation strategies to enhance the functionality of recombinant lactobacilli. This engineered living material approach is promising for developing L. plantarum towards advanced applications in biotechnology, pharmaceutics, and living therapeutics.Competing Interest StatementThe authors have declared no competing interest.