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    Improved gene expression in E. coli

    products-servicesenGenes Biotech GmbH
    February 13th 2020

    enGenes Biotech has developed customizable and scalable solutions for the production of recombinant proteins in microbial expression systems.

    Its world-leading enGenes-X-press technology delivers outstanding gene expression in Escherichia coli (E.coli). This platform technology for growth-decoupled protein production allows enGenes to develop economic, scalable and efficient production processes within short time frames to make clients’ unique products into commercial successes.

    Cost-effective prokaryotic protein expression

    enGenes combines in-depth experience in cutting-edge genetic engineering with a broad expertise in bioprocess engineering to deliver best-in-class microbial protein expression systems. Based on a strong understanding for the requirements of the biotech industry and extensive knowledge and method portfolio, enGenes delivers improved host cell lines, plasmid expression vectors and optimized production processes to maximize bio-process efficacy.

    Its customized project plans and feasibility studies also responds to individual documentation requirements to support an easy transfer to clients’ own facilities and quality management systems. enGenes was founded by internationally recognized scientists in the field of microbial-based protein expression with an extensive track-record of scientific publications, patents and a strong understanding for the requirements of the biotech industry from a long history of international cooperation.

    Optimizing gene expression in Escherichia coli

    enGenes offers a modular system of customer services for gene expression and protein production in Escherichia coli and other prokaryotes (e.g.Bacillus subtilis). Our service, focused on the expression of difficult-to-express proteins, is divided into a series of steps which can be performed either individually or in combination – so you can decide which of the steps you actually need.

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

    The following service options are available:

    • Cloning / Vector optimization
    • Antibiotic resistance-free plasmid backbones available
    • Genetic engineering of host cell lines
    • Feasibility studies for our proprietary enGenes-X-press technology
    • Upstream-Process Development
    • Downstream-Process Development
    • Non-GMP recombinant protein production
    • Non-GMP plasmid DNA production
    • Tech transfer to CMO or plant construction/engineering company
    • Consulting to achieve most cost effective manufacturing solution

    enGenes X-press gene expression features

    X-press is an unique and proprietary growth decoupled protein production platform that allows customers substantial yield improvements and COGS reductions.

    The X-press 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.

    The technology allows secretion of proteins targeted to the periplasmic space to the cell free supernatant, thereby allowing a cost-effective manufacturing option comparable to yeast-based expression systems. It also enables recombinant product formation to be decoupled from cell growth, enabling bioprocessing with clear separation of biomass growth and product formation. 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).

    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.

    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
    Improved gene expression in E. coli

    Schematic of enGene-X-press production process

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