The Need: In the realm of biotechnology, the challenge of synthesizing enzymes, pharmaceuticals, and fine chemicals efficiently is paramount. However, the presence of multiple proteases secreted by Bacillus subtilis 168 during growth stages poses a significant obstacle to achieving optimal production yields. Overcoming this limitation is crucial for enhancing the productivity and scalability of bioprocesses utilizing B. subtilis as a production platform.

The Technology: Introducing B. subtilis KO7, a breakthrough seven-protease deletion strain meticulously engineered to address the limitations of wild-type strains. Constructed from the widely used laboratory host, PY79, KO7 undergoes sequential knockout of protease genes without incorporating antibiotic resistance genes or integrated plasmids. Each knockout is validated through sequencing, ensuring marker-free, in-frame deletions with minimal genetic alterations. KO7 exhibits rapid growth in standard minimal media, offering a robust and versatile platform for bioproduction applications.

Commercial Applications:

• Industrial enzyme synthesis
• Pharmaceutical production
• Fine chemical manufacturing
• Bioprocess optimization
• Bioremediation applications

Benefits/Advantages:

• Enhanced productivity: Elimination of protease secretion optimizes production yields.
• Simplified downstream processing: Marker-free deletions reduce purification complexity.
• Cost-effective production: Efficient growth in standard media minimizes resource expenditure.
• Versatile platform: Suitable for diverse bioproduction applications.
• Open access: Available in the public domain with no downstream rights restrictions, fostering innovation and collaboration.