E. coli Expression System

Cost-effective and scalable ncAA incorporation

Non-canonical amino acids (ncAAs) are powerful tools in modern protein engineering, drug discovery, and fundamental biological research. Our service leverages the robust E. coli expression system to enable efficient, site-specific incorporation of ncAAs into target proteins, providing researchers with a reliable, versatile, and cost-effective solution to expand the chemical and functional diversity of proteins.

Advanced Technology Platform

We engineer the translational machinery of E. coli by integrating an orthogonal aminoacyl-tRNA synthetase (aaRS) and a matching tRNA pair. This system reassigns a specific stop codon—commonly the amber codon UAG—to incorporate the chosen ncAA with high precision. The orthogonal pair operates independently of the host's native protein synthesis, ensuring accurate placement of the ncAA without affecting overall cellular function.

Key Advantages

High Efficiency & Yield – Optimized codon usage, regulatory elements, and tailored culture conditions maximize both ncAA incorporation and protein expression.
Broad ncAA Compatibility – Supports a wide range of ncAAs for applications such as bio-orthogonal labeling, improved protein stability, or enhanced activity.
Scalable Production – Produces high-purity proteins from milligram to gram scale, suitable for structural biology, biochemical assays, and other research needs.
End-to-End Service – Covers everything from gene design to purified protein, ensuring smooth project execution.

Service Workflow

1) Project Consultation – Define your target protein, desired ncAA, and application objectives.

2) Gene Design & Plasmid Construction – Design the gene sequence and assemble the expression vector with the orthogonal system.

3) Expression Optimization – Screen and select the most suitable E. coli strains and culture conditions to maximize protein yield.

4) Protein Production & Purification – Perform large-scale expression and purification to achieve the desired purity and quantity.

5) Quality Verification – Confirm ncAA incorporation and protein integrity using mass spectrometry, Western blot, and other analytical techniques.

Applications

Drug Discovery – Generate protein variants for therapeutic screening and development.
Biosensors – Engineer proteins with enhanced detection capabilities.
Structural Biology – Produce labeled proteins for crystallography, NMR, and cryo-EM studies.
Functional Studies – Introduce novel chemical functionalities to probe protein mechanisms.

Our E. coli ncAA incorporation service combines precision, scalability, and flexibility, enabling researchers to explore new dimensions in protein design and application.