Synthetic Biology Toolbox for Lactiplantibacillus plantarum: Engineering Next-Generation Probiotic Cell Factories.

Lactiplantibacillus plantarum has emerged as a promising synthetic biology chassis owing to its food-grade safety, probiotic properties, and adaptability to the gastrointestinal tract. Its engineering potential has expanded rapidly in live biotherapeutics, functional foods, biosensors, and animal health. However, its broader application is still limited by the lack of standardized genetic parts, versatile engineering tools, and industrially compatible chassis. This review summarizes recent advances in the synthetic biology toolbox of L. plantarum, including expression elements, genome engineering strategies, and representative applications. Particular emphasis is placed on the development of genetic components, conventional and CRISPR-based engineering technologies, and their applications in medicine, food biotechnology, biosensing, and animal nutrition. Current challenges and future opportunities, including standardized genetic parts, precision genome engineering, AI-assisted strain design, and high-performance chassis development, are also discussed. This review provides a comprehensive framework for developing next-generation programmable probiotic cell factories based on L. plantarum.