Recent advances in genetic tools for engineering probiotic lactic acid bacteria

Abstract

Synthetic biology has grown exponentially in the last few years, with a variety of biological applications. One of the emerging applications of synthetic biology is to exploit the link between microorganisms, biologics, and human health. To exploit this link, it is critical to select effective synthetic biology tools for use in appropriate microorganisms that would address unmet needs in human health through the development of new game-changing applications and by complementing existing technological capabilities. Lactic acid bacteria (LAB) are considered appropriate chassis organisms that can be genetically engineered for therapeutic and industrial applications. Here, we have reviewed comprehensively various synthetic biology techniques for engineering probiotic LAB strains, such as clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 mediated genome editing , homologous recombination, and recombineering. In addition, we also discussed heterologous protein expression systems used in engineering probiotic LAB. By combining computational biology with genetic engineering, there is a lot of potential to develop next-generation synthetic LAB with capabilities to address bottlenecks in industrial scale-up and complex biologics production. Recently, we started working on Lactochassis project where we aim to develop next generation synthetic LAB for biomedical application.

Next-Generation Probiotics as Novel Therapeutics for Improving Human Health: Current Trends and Future Perspectives

“…Probiotics can have a variety of effects on the host ( Figure 2 ), including changes in the gut microbiome, modulation of the immune system, and production of bioactive compounds. The mechanisms of action of the NGPs on gut health, immune function, and other health outcomes can vary depending on the specific strain or formulation [ 53 , 62 ]. However, probiotics have a wide range of beneficial effects on gut health and the immune system.…”

“…Probiotics can also modulate the immune system by influencing the activity of immune cells (T-cells and B-cells) and regulating the inflammatory response. Additionally, they can help strengthen the gut barrier, preventing harmful substances from entering the body and enhancing the absorption of nutrients, particularly in the small intestine [ 53 , 62 , 65 ]. According to recent reports, the Faecalibacterium prausnitzii showed ability to induce the anti-inflammatory cytokine IL-10 in peripheral blood mononuclear cells which is likely due in part to their production of short-chain fatty acids or butyrate [ 66 ].…”

Therapeutic Applications of Native and Engineered Saccharomyces Yeasts

“…In addition, CRISPR/Cas9-based genomic integration of the human lysozyme secretion cassette rendered the Sb culture supernatant capable of lysing bacteria [14]. CRISPR/Cas9-based genomic integration is a more appropriate approach than conventional yeast cloning methodologies for building synthetic probiotics whose biological functions must be stably and accurately manifested in the host gut environment without unnecessary heterologous genetic elements including antibiotic resistance markers [67]. This study successfully proved the potential of CRISPR-based molecular tools and metabolic engineering strategies for developing yeast-based avenues for controlling digestive conditions.…”

Expanding the genetic programmability of Lactiplantibacillus plantarum

“…A repertoire of promoters is crucial for establishing synthetic genetic circuits in a microbial chassis (Mays & Nair, 2018 ; Mugwanda et al, 2023 ). Different strategies have been pursued to increase the available set of promoters for L. plantarum , like screening native promoters from the bacterial genome, creating synthetic promoter libraries by random or directed mutagenesis and employing promoter regions from both phylogenetically related and distant bacterial species.…”