CD Biosynsis offers specialized Protein Expression and Purification Services utilizing the versatile Pseudomonas putida platform. P. putida serves as an excellent intermediate host, bridging the gap between simple prokaryotes (like E. coli) and complex eukaryotes. It possesses superior features such as high respiratory capacity, intrinsic robustness, and suitability for complex protein folding and production of toxic metabolites. Our service covers the entire workflow: gene synthesis and vector construction, optimized fermentation, and multi-step purification (chromatography), ensuring the delivery of high-yield, high-purity, and biologically active recombinant proteins, particularly those requiring specific redox conditions or post-translational modifications (PTMs) typical of aerobic systems.
Get a QuoteRecombinant protein production often faces challenges such as low yield, poor solubility, and incorrect folding when using standard hosts like E. coli, especially for complex enzymes, cytochrome P450s, or proteins involved in redox processes. P. putida offers a superior oxidative environment and sophisticated membrane systems, making it highly effective for these demanding proteins. Our platform is optimized to exploit P. putida's unique metabolic features, using tailored expression vectors and fermentation strategies (e.g., high-cell-density culture) to dramatically increase the expression level of your target protein while maintaining its native conformation and biological activity.
Aerobic Environment
P. putida’s efficient aerobic metabolism supports better disulfide bond formation and protein folding, reducing aggregation into inactive inclusion bodies.
Lower Growth Stress
Compared to fast-growing E. coli, controlled growth in P. putida allows more time for proper protein maturation and quality control.
Robust Physiology
High tolerance to temperature, pH fluctuations, and high concentrations of metabolic products, ideal for large-scale fermentation.
Efficient Biocatalysis
Superior respiratory chain and energy generation capacity support the high energy demands of expressing large amounts of recombinant protein.
P450 Co-expression
P. putida can successfully co-express necessary reductases and electron transport components for active cytochrome P450 enzymes.
Specific Post-Translational Modifications (PTMs)
Capability to perform specific PTMs not available in standard E. coli strains, enhancing biological relevance.
Our comprehensive service ensures a seamless transition from gene sequence to purified, active protein.
1. Gene & Vector Optimization
2. Strain Transformation & HCDC
3. Cell Lysis & Extraction
4. Multi-Step Purification
Codon optimization for maximal expression in P. putida.
Construction of custom expression vectors (e.g., based on PTL or pSEVA systems) with strong, inducible promoters (e.g., Ptet, Prha).
Addition of purification tags (His-tag, GST, Strep-tag) and cleavage sites (e.g., TEV protease).
Transformation of the expression vector into the target P. putida chassis (e.g., KT2440 derivatives).
Optimization of High-Cell-Density Culture (HCDC) fermentation parameters (media, induction time, temperature, pH) for maximum yield.
Affinity Chromatography (e.g., IMAC for His-tagged proteins) for initial capture and high-fold enrichment.
Polishing steps: Size Exclusion Chromatography (SEC) or Ion Exchange Chromatography (IEX) to achieve high purity (typically >95%).
QC: SDS-PAGE and Western Blot for purity, and MS/Activity Assay for functional verification.
High-Copy Inducible Vectors
Use of robust, broad-host-range plasmids designed to maintain stability and high copy number in P. putida under selection.
Automated HCDC Fermentation
Precise monitoring and control of Dissolved Oxygen (DO), feeding rate, and pH in bioreactors to ensure optimal cell viability and protein yield.
Co-factor Supply Optimization
Media supplementation or pathway engineering to ensure sufficient supply of critical cofactors (e.g., heme, FAD, NAD(P)H) for complex enzymes.
Got questions about your protein project?
Contact UsWhat types of proteins benefit most from P. putida expression?
Proteins that are typically difficult to express actively in E. coli, such as complex multi-domain enzymes, redox-dependent proteins (e.g., Cytochrome P450s), and certain membrane proteins, benefit significantly from P. putida’s robust aerobic system.
What is the typical purity level guaranteed?
We typically guarantee a minimum purity of 95% for research-grade proteins, with the capability to achieve >98% homogeneity depending on the client’s final application requirements (e.g., crystallography).
Do you offer endotoxin removal?
Yes. As P. putida is a Gram-negative bacterium, endotoxin removal is a standard option. We use specialized chromatography (e.g., endotoxin removal resin) to ensure the final product meets low endotoxin requirements for cellular or animal studies.
What information is required to start a project?
We primarily need the target protein sequence (amino acid or nucleotide), the desired purity level, the final yield goal, and any known specific requirements (e.g., inclusion of cofactors, specific buffer conditions).
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.