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Large-scale purification of enzymes.

C J Bruton

    Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
    |January 26, 1983
    PubMed
    Summary

    Scaling up protein purification requires different methods than lab-scale techniques. Enzymes from thermophilic sources offer advantages for large-scale production due to better storage properties and tractability.

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    Area of Science:

    • Biochemistry
    • Biotechnology
    • Chemical Engineering

    Background:

    • Laboratory-scale protein purification methods often fail when scaled up.
    • Large-scale enzyme production necessitates consideration of storage stability and handling properties.

    Purpose of the Study:

    • Evaluate economic advantages of simultaneous multi-enzyme isolation versus individual purification.
    • Identify scalable purification techniques for producing gram to kilogram quantities of enzymes.

    Main Methods:

    • Batch adsorption/elution using ion-exchange celluloses replaces precipitation.
    • Hollow-fibre ultrafiltration replaces dialysis for concentration/dilution.
    • Column chromatography is scaled up, with modifications for flow rate optimization.
    • Triazine dye conjugates offer a cost-effective quasi-affinity chromatography alternative.

    Main Results:

    • Comparable purification factors and recoveries are achievable at large scale.
    • Triazine dye matrices are inexpensive, easy to prepare, and stable.
    • Scaling up column chromatography is generally feasible with geometric modifications.

    Conclusions:

    • Large-scale enzyme purification benefits from adapted techniques like ultrafiltration and triazine dye chromatography.
    • Thermophilic enzymes are advantageous for large-scale applications due to stability.
    • Simultaneous multi-enzyme isolation may offer economic benefits.

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