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Process integration of cell disruption and fluidised bed adsorption of microbial enzymes: application to the retro-design of the purification of L-asparaginase

Bierau, Horst (2001)
Ph.D. thesis, University of Birmingham.

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The practical feasibility and generic applicability of the direct integration in the same time frame of cell disruption by bead milling with the capture of intracellular products by fluidised bed adsorption has been demonstrated. Pilot-scale purification of the enzyme L-asparaginase from unclarified Erwinia chrysanthemi disruptates exploiting this novel approach yielded an interim product which rivalled or bettered that produced by the current commercial process employing discrete operations of alkaline lysis, centrifugal clarification and batch adsorption. In addition to improved yield and quality of product, the process time during primary stages of purification was greatly diminished. Two cation exchange adsorbents, CM HyperD LS (Biosepra/Life Technologies) and SP UpFront (custom made SP form of a prototype stainless steel/agarose matrix, UpFront Chromatography) were physically and biochemically evaluated for such direct product sequestration. Differences in performance with regard to product capacity and adsorption/desorption kinetics were demonstrated and are discussed with respect to the designof adsorbents for specific applications. In any purification of L-asparaginase (pI=8.6), product-debris interactions commonly diminish the recovery of available product. It was demonstrated herein, that immediate disruptate exposure to a fluidised bed adsorbent promoted concomitant reduction of product in the liquid phase, which clearly counter-acted the product-debris interactions to the benefit of overall product yield.

Type of Work:Ph.D. thesis.
Supervisor(s):Lyddiatt, Andrew
School/Faculty:Schools (1998 to 2008) > School of Engineering
Department:Chemical Engineering
Subjects:TP Chemical technology
Institution:University of Birmingham
Library Catalogue:Check for printed version of this thesis
ID Code:71
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
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