B. subtilis, being a gram-positive endospore forming bacteria, is the most desirable and favorable choice for cell surface display with spore coat protein as anchoring proteins, including CotB, CotC, CotE, CotG, CotX, CotY, CotZ, CgeA, and OxdD. For successful spore surface display, it is significant to select an appropriate anchor protein because it contains efficient signal peptide or transporting signal for efficient promotion of transportation, immobilization and stability. B. subtilis spore surface display, a well-established technology for protein analysis and engineering, involves expressing exogenous proteins as a genetic fusion to either the N- or C-terminus of anchoring proteins.
Fig.1 Schematic representation of B. subtilis surface display systems
• They must have a strong anchoring domain to ensure that exogenous proteins can be immobilized on the spore surface.
• They must be compatible with foreign proteins, be able to form fusion proteins, and should not be able to interact with each other.
• They must be resistant to protease hydrolysis.
• Do not need to cross the membrane barrier.
• Stress Tolerance:heat resistance, alkaline tolerance, organic solvent and enzymatic resistance, ultraviolet radiation resistance
• Easy recovery
• Well-established fermentation and production technology
• GRAS (nonpathogenicity)
Hence, spore surface display is the most desirable and the most promising technology which provides feasible avenues to improve industrial production efficiency. Kwon et al. (2007) developed a new kind of whole-cell biocatalyst which display β-galactosidase on B.subtilis spore by fusion it to the spore CotG protein.
• Vaccine, Antigen, Antibody, Pharmaceutical Protein
• Enzyme Immobilization/Biocatalysis
• Peptide library screening
• Environmental bioadsorbents
• We have many years of service experience in cell surface display.
• We have a world-class service platform.
• The price of Creative Biogene is very competitive in the industry and the price is favorable.
• One-stop service.
• Ensure efficient service, and urgent service can be provided.
• Periodic progress updates.
• SDS-PAGE Analysis
• Western Blot Analysis
• Enzyme assays
• Fluorescence assays
• Flow cytometry
• Immunofluorescence microscopy
In general, our turnaround time is 4-8 weeks depending on the size of your project.
Creative Biogene develops the recombinant approach which is based on the modification of B. subtilis genome to display a passenger protein of interest as a fusion with spore coat protein. At present, we select the non-essential genes amyE, lacA and pyrD as insertion sites to creating a multi-anchoring protein display system. With the rich experience, our talented scientists advise appropriate construction strategy in various biotechnological applications. Please feel free to contact us if you have any concerns.
1. Chen H., ·Ullah J., & ·Jia J. (2017). Progress in bacillus subtilis spore surface display technology towards environment, vaccine development, and biocatalysis. J Mol Microbiol Biotechnol.
2. Lin, P. , Yuan, H. , Du, J. , Liu, K. , & Wang, T. . (2020). Progress in research and application development of surface display technology using bacillus subtilis spores. Applied Microbiology and Biotechnology, 104(6), 2319-2331.