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Computer-based simulation of the recovery of intracellular enzymes and its pilot-scale verification

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Abstract

Accurate and reliable models are required for a range of unit operations if simulations are to be used for accelerating the design and optimisation of bioprocesses. This paper presents results of pilot-plant studies that have been used to verify process simulations for a sequence of operations comprising of cell disruption, fractional protein precipitation and centrifugal separation. These have been tested using the purification of the enzyme alcohol dehydrogenase (ADH) from Saccharomyces cerevisiae as being representative of the recovery of a labile intracellular enzyme. Comparison of pilot-plant against simulated data highlights where improvements to the models are required and has resulted in increased confidence in the simulations for a wide range of conditions including the operational scale and the nature of the starting material. Results demonstrate the effectiveness of the verification approach for the development of reliable predictive models to assess the feasibility of process designs and performance of a train of bioprocess operations.

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Abbreviations

ADH:

Alcohol dehydrogenase (EC 1.1.1.1)

DNA:

Deoxyribonucleic acid

NAD+ :

Nicotinamide adenine dinucleotide

D :

Agitator diameter (m)

d :

Diameter of precipitate particles (μm)

G :

Average shear rate (s−1)

g :

Gravity constant (9.81 ms−2)

k :

Kinetic constant (pass−1)

Q :

Flowrate (m3s−1)

Σ :

Equivalent area of settling for a centrifuge (m2)

S :

Standard deviation of precipitate particle size distribution (μm)

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Authors and Affiliations

  1. Bio-Products Laboratory, Dagger Lane, WD6 3BX, Elstree, Herts

    M. Bulmer

  2. Jacobs Engineering, Dale House, SK1 1TA, Tiviotdale, Stockport, Cheshire

    A. I. Clarkson

  3. The Advanced Centre for Biochemical Engineering, Dept. of Chemical andBiochemical Engineering, University College London, Torrington Place, WC1E 7JE, London, UK

    N. J. Titchener-Hooker & P. Dunnill

Authors
  1. M. Bulmer
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  2. A. I. Clarkson
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  3. N. J. Titchener-Hooker
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  4. P. Dunnill
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Additional information

UCL is the Biotechnology and Biological Sciences Research Council's sponsored Advanced Centre for Biochemical Engineering and the Council's support is gratefully acknowledged. The authors would like to thank I Ms. N. Abidi for technical assistance.

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Bulmer, M., Clarkson, A.I., Titchener-Hooker, N.J. et al. Computer-based simulation of the recovery of intracellular enzymes and its pilot-scale verification. Bioprocess Engineering 15, 331–337 (1996). https://doi.org/10.1007/BF02426444

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