PT  - JOURNAL ARTICLE
AU  - Blanch-Asensio, Marc
AU  - Tadimarri, Varun Sai
AU  - Martinez, Roberto
AU  - Dahiya, Gurvinder Singh
AU  - Lale, Rahmi
AU  - Sankaran, Shrikrishnan
TI  - Genetic and materials engineering to enhance inducible gene expression in lactobacilli
AID  - 10.1101/2025.01.10.632477
DP  - 2025 Jan 01
TA  - bioRxiv
PG  - 2025.01.10.632477
4099  - http://biorxiv.org/content/early/2025/01/13/2025.01.10.632477.short
4100  - http://biorxiv.org/content/early/2025/01/13/2025.01.10.632477.full
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.