Improved recombinant protein production in E. coli

enGenes Biotech has developed its unique enGenes-e^Xpress^™ proprietary technology platform for outstanding recombinant protein production in Escherichia coli (E.coli). -e^Xpress uses an advanced growth decoupled mechanism platform that allows customers to achieve customized solutions for the cost-effective production of recombinant proteins in microbial expression systems.

The technology is leveraged from enGenes’ world-class experience in cutting-edge genetic engineering, combined with a broad expertise in bioprocess engineering, to produce a range of best-in-class protein expression systems.

enGenes-e^Xpress^™ features

The -e^Xpress technology platform is based on a genetically modified host cell based on BL21 (DE3) that allows introduction of standard expression vectors (with T7 promoter) with no need for cumbersome adaptations. The fermentation process can be implemented with standard fermentation equipment.

Reprogramming of the host cell is performed by co-expression of a bacteriophage-derived peptide that stops cell division and host mRNA production and at the same time modulates the host cell metabolism for improved soluble, high level protein production.

The technology enables recombinant product formation to be decoupled from cell growth, enabling bioprocessing with clear separation of biomass growth and product formation. It also allows secretion of proteins targeted to the periplasmic space to the cell free supernatant, thereby opening up cost-effective manufacturing options comparable to yeast-based expression systems. This in turn makes it possible to generate significantly higher specific and volumetric yields compared to the previous state-of-the-art (Escherichia coli BL21 (DE3).

Observed yields include:

• Glycosyltransferase = 1 g/L (cytoplasm)
• Affinity ligand = 8-10 g/L (cell supernatant)
• Soluble enzyme (hydrolase) = 13 g/L (cytoplasm)
• VHH (variable camelid heavy chain-only) single domain antibody = 1 g/L (cell supernatant)
• Antibody fragment = 250 mg/L (cell supernatant)
• Green fluorescent protein (GFP) = 20 g/L

enGenes-e^Xpress applications

enGenes offers a modular system of customer services for gene expression and protein production in E. coli and other prokaryotes. The enGenes service, focused on the expression of difficult-to-express proteins, is divided into a series of steps that can be performed either individually or in combination – allowing customers to select only those steps that are actually needed.

These services are designed to adequately cover all specific demands along the line from gene to protein, principally:

• Cloning and vector optimization
• Generation of expression strain and host cell engineering
• Feasibility studies for our proprietary enGenes-e^Xpress technology
• Lab scale fermentations and bioprocess optimization
• Establishing of purification protocol and final purification
• Up-scaling and tech-transfer to Contract Manufacturing Organizations (CMOs)

Improved recombinant protein production in E. Coli

-e^Xpress technology for growth-decoupled protein production in E. coli allows the user to develop economic, scalable and efficient production processes within short time frames to make your unique product a commercial success.

The enGenes-e^Xpress protein production platform is defined by a number of unique characteristics:

• E. coli based, growth decoupled production system
• Allowing decoupling of protein/DNA production from cell growth
• Host cell growth halted at defined cell concentration
• Protein of interest production triggered
• Host cell resources used exclusively for product of interest
• Proprietary IP (PCT/EP2016/059597)