CD Biosynsis delivers comprehensive Bacillus subtilis Protein Expression and Purification Services, leveraging this robust Gram-positive bacterium for the high-yield, large-scale production of diverse proteins, enzymes, and therapeutic peptides. Bacillus subtilis is an excellent host, particularly suited for secreted protein expression, avoiding the inclusion body formation often seen in E. coli. Our platform integrates state-of-the-art gene synthesis, expression vector optimization, and advanced chromatography techniques. We offer end-to-end solutions, from codon optimization and promoter screening to multi-step purification, ensuring the final protein product achieves the required high purity (>95%) and biological activity for your downstream applications, including structural biology and drug development.
Get a QuoteBacillus subtilis is widely preferred over E. coli for producing many types of recombinant proteins due to its powerful capacity for protein secretion directly into the culture medium. This capability simplifies purification and ensures correct protein folding, as the protein avoids the reducing environment and crowding of the cytoplasm. As a non-pathogenic (GRAS status) host, B. subtilis is especially suitable for producing food-grade enzymes and certain therapeutics. Our expertise focuses on optimizing secretion pathways and utilizing highly efficient, regulated promoters to maximize protein yield and solubility.
Gene Synthesis & Codon Optimization
Synthetic gene design, including optimization of the codon usage frequency and mRNA secondary structure for high expression in B. subtilis.
Expression Vector Construction
Selection and construction of high-copy number plasmids or stable chromosomal integration vectors with strong, regulated promoters (e.g., P43, Pgrac).
Secretion Signal Optimization
Testing and selection of optimal secretion signal peptides (e.g., AmyQ, SP-lip) to maximize protein translocation efficiency into the medium.
Custom Host Strain Selection
Selection of protease-deficient strains (e.g., WB800N) or genetically optimized strains to reduce degradation and improve target protein stability.
Fermentation Optimization
Optimization of culture conditions (media, temperature, induction kinetics) for high-density fermentation in shake flasks up to bioreactors.
Process Scale-Up
Efficient transition from lab-scale (mg) to pilot-scale (gram) production while maintaining optimal yield and quality.
Multi-Step Chromatography
Utilizing techniques like Affinity Chromatography (Ni-NTA, Strep-Tactin), Ion Exchange (IEX), and Size Exclusion Chromatography (SEC) for high purity (>95%).
Endotoxin Removal
Specific protocols for low endotoxin levels (typically < 1 EU/mg) required for in vivo or cell-based assays.
Purity and Activity QC
SDS-PAGE and Western Blot for purity; Mass Spectrometry (MS) for confirmation; and Custom Functional Assays for activity verification.
Our systematic approach ensures predictable timelines and consistently high-quality protein delivery.
1. Gene Design & Vector Prep
2. Host Transformation & Strain Selection
3. Expression & Fermentation
4. Purification & Final QC
Codon optimization for maximum B. subtilis expression efficiency.
Design of appropriate secretion signals, purification tags (His-tag, GST), and cleavage sites.
Construction of the final B. subtilis expression vector.
Transformation of the expression vector into the selected B. subtilis host strain.
High-throughput screening and selection of the highest-yielding clone.
Verification of integration/plasmid presence and preliminary expression.
Target protein isolation using multi-step chromatography.
Concentration, buffer exchange, and sterile filtration.
Final quality analysis (Purity >95%) and delivery of the Certificate of Analysis (CoA).
Superior Secretion Capability
Leverage the native high-capacity secretion system to yield soluble proteins directly in the medium, minimizing refolding complexity.
Guaranteed High Purity (>95%)
Use sophisticated multi-step chromatography (IEX, SEC, Affinity) to deliver highly pure proteins suitable for structural studies.
Scalable and Cost-Effective
Proven protocols for high-density fermentation, allowing seamless and cost-effective scale-up from milligram to gram quantities.
Low Endotoxin Levels
As a Gram-positive host, B. subtilis naturally produces lower endotoxin, and we offer optional endotoxin removal for therapeutic applications (< 1 EU/mg).
"We struggled to express a key enzyme solubly in E. coli. Switching to CD Biosynsis's B. subtilis platform resulted in high yields of active, secreted protein. The purity was excellent for our crystallization studies."
Dr. Chen, Principal Investigator, Structural Biology Lab
"The scale-up process was flawless. They took our pilot project and delivered a gram of high-purity protein on time and within budget. Their communication throughout the fermentation optimization phase was outstanding."
Mr. David Smith, Project Manager, Industrial Enzyme Manufacturer
"The low endotoxin delivery of our therapeutic peptide expressed in B. subtilis was critical. The final product passed all our pre-clinical safety tests immediately."
Dr. Lena Koo, R&D Scientist, Pharmaceutical Development Group
"Their optimization of the secretion signal made the difference. We achieved twice the yield compared to our previous internal attempts using a different Gram-positive host. The final product was functionally validated."
Dr. Alan Rivas, Lab Director, Applied Microbiology Institute
Why choose B. subtilis over E. coli for protein expression?
B. subtilis is superior for proteins requiring proper folding and disulfide bond formation. Its secretion system releases the protein into the medium, simplifying purification and avoiding the challenge of inclusion bodies common in E. coli.
What is the minimum guaranteed purity level for purified proteins?
We guarantee a minimum purity of >95% for most recombinant proteins, achieved through multi-step chromatography (Affinity, IEX, SEC). Higher purity (>98%) can be requested for demanding applications.
Can you express large proteins or multi-subunit complexes?
Yes, the B. subtilis system is capable of expressing large proteins. For multi-subunit complexes, we use co-expression strategies and co-purification protocols to ensure the integrity and correct stoichiometry of the final assembly.
How do you minimize protein degradation during expression?
We primarily use protease-deficient host strains (e.g., those lacking six major proteases) and optimize fermentation conditions (temperature, harvest time) to significantly reduce non-specific protein degradation.
What are the typical endotoxin levels for the final purified protein?
Due to its Gram-positive nature, B. subtilis naturally has lower endotoxin contamination. For therapeutic use, we offer specialized polishing steps to achieve endotoxin levels below 1 EU/mg.
Do you offer a functional assay to verify protein activity?
Yes, in addition to standard purity QC (SDS-PAGE, MS), we offer customized functional assays (e.g., enzyme kinetics, binding assays) designed in collaboration with the client to confirm the biological activity of the purified protein.
What production scales can you handle?
We handle production scales ranging from small-scale milligram (mg) production for initial research to large-scale gram production in high-density bioreactors for commercial applications.
How is the expression vector integrated into the host?
We offer both high-copy plasmid-based expression for high flexibility and stable chromosomal integration using gene editing for long-term genetic stability, depending on the client's needs.
CRISPR-Cas9 technology represents a transformative advancement in gene editing techniques. The main function of the system is to precisely cut DNA sequences by combining guide RNA (gRNA) with the Cas9 protein. This technology became a mainstream genome editing tool quickly after its 2012 introduction because of its efficient, simple and low-cost nature.
The CRISPR gene editing system with its Cas9 version stands as a vital instrument for current biological research. CRISPR technology enables gene knockout (KO) through permanent gene expression blockage achieved by sequence disruption. Various scientific domains including disease modeling and drug screening employ this technology to study gene functions. CRISPR KO technology demonstrates high efficiency and precision but requires confirmation and verification post-implementation because unsatisfactory editing may produce off-target effects or incomplete gene knockouts which impact experimental result reliability. For precise and efficient Gene Editing Services - CD Biosynsis, Biosynsis offers comprehensive solutions tailored to your research needs.
The CRISPR-Cas9 knockout cell line was developed using CRISPR/Cas9 gene editing to allow scientists to remove genes accurately for research on gene function and disease models and pharmaceutical discovery. Genetic research considers this technology essential due to its high efficiency together with simple operation and broad usability.
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CD Biosynsis is a leading customer-focused biotechnology company dedicated to providing high-quality products, comprehensive service packages, and tailored solutions to support and facilitate the applications of synthetic biology in a wide range of areas.