Are you looking for an endotoxin-free expression system? Opt for a safe E.coli alternative with our Bacillus subtilis expression system. Benefit now from a tailored protein expression
protocol thanks to a pilot study ahead of large-scale manufacturing.

After selecting the optimum conditions, further purification tests are performed to figure out the highest possible recombinant protein yields. The protocol is then up-scaled to 1L protein production using the selected conditions.

A difficult-to-express protein? Optimize your chances to express it thanks to our high throughput protein expression screening service.

Case Study : High Throughput Expression Protein Screening in Bacillus Subtilis

Aim of the project:

Our customer requested the production of an insecticidal protein from B.thuringiensis. This protein is difficult to express and to obtain in a soluble recombinant form. This project aims to determine an optimum protocol for this protein production using High Throughput Expression Protein Screening (HTP) in Bacillus subtilis.

Protein expression tests:

To figure out the optimum condition for protein expression, the client consulted our high throughput expression screening service. Several parameters were examined during this protocol:

• Different B.subtilis strains.
• Gene design including codon optimization.
• Different expression strategies.
• Several induction conditions.

A total of 72 expression tests are conducted, and a total of 144 analyzed samples.

More details about the protocol are provided in the complete PDF report.

Small-scale production of the best expression combination:

Conclusion of the HTP expression screening in Bacillus subtilis:

Thanks to high throughput expression screening, the protein can now be produced in native and soluble conditions with an expression yield of up to ~30mg/L in standard conditions.

Why choose Bacillus Subtilis for your Protein Expression?

Belonging to Bacillus species, Bacillus subtilis is a well-studied gram-positive bacterium. It has become a popular protein factory host for industrial manufacturing. It presents significant attributes in comparison with gram-negative bacteria such as E.coli.

•  B.subtilis Protein Expression as an Alternative to E.coli:

In terms of large-scale production, Bacillus subtilis constitutes an easy and inexpensive organism to culture. As it is genetically well characterized, it simplifies genetic manipulation. Plus, it is known for its rapid proliferation rates.

Furthermore, Bacillus subtilis is recognized as safe. It acquired the GRAS status (Generally Recognized as Safe) by the US Food and Drug Administration.

• Simple Downstream Processing:

Thanks to its efficient secretory pathways, B.subtilis can export expressed proteins directly into the culture media. Newly synthesized proteins can acquire proper folding and prevent protein accumulation and inclusion body formation. Therefore, the cell naturally avoids intracellular toxicity by isolating proteins in the extracellular environment. Unlike gram-negative bacteria, B.subtilis are deprived of endotoxins. This simplifies the downstream process in cases where the final product must be endotoxin-free.

• Applications of Bacillus Subtilis as an Expression System:

Bacillus subtilis occupies an important place in several areas, including the medical, agro-food, and ecological fields. It constitutes a cell factory for therapeutical heterologous protein proteins, amino acids, bioactive molecules, and vaccines.

It is also the preferred prokaryotic organism in the biopharmaceutical and agro-food fields. 60% of enzyme production in the industry is established using B.subtilis such as proteases, amylases, lipases, and galactosidase. Furthermore, human interferon-β/ γ and interferon-α 2b and other membrane proteins are better produced in B.subtilis than in E.coli. Even though protein yields are not as high as E.coli, better protein folding is accomplished with B.subtilis. thanks to their slower translation rates.

Please feel free to contact us for further information!