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Process integration of the initial recovery and purification stages for multiple enzyme production.

Beer, Simon; (1991) Process integration of the initial recovery and purification stages for multiple enzyme production. Doctoral thesis (Ph.D.), University College London. Green open access

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Abstract

The aim of this project was to quantify the relationships between the fermentation and the early stages of the fermenter broth processing, and the performance of the initial purification stage, namely fractional precipitation. In addition the recovery of multiple products from a single fermentation broth was appraised. Fractionation diagrams were developed as a means of evaluating the performance of two cut precipitation procedures especially where the first and second cuts were highly Interactive, i.e. the operation of the first cut determined the subsequent fractionation. The system used examined the recovery of two enzymes, glucose oxidase and catalase produced intracellularly by the growth of Aspergillus niger in stirred tank fermenters on scales between 30L and 4000L. The primary recovery operations consisted of cell recovery in a basket centrifuge, washing by resuspension and disruption in a Man ton Gaul in homogeniser. The removal of cell debris and the purification by fractional precipitation using propan-2-ol was on a bench scale. The propan-2-ol was added using a gradient mixer to allow sampling of the protein-precipitant mix at varying concentrations with the history of the material being close to that experienced in a batch stirred tank. The titre of glucose oxidase, catalase and total protein were measured in the supernatant once precipitate had been removed in a centrifuge. The results indicate that since none of the primary recovery operations caused major differences in the Initial purification all non-essential operations could be removed. However in a simulation of the possible consequences of genetic manipulation the effect of the relative levels of the different products produced major differences in the purification. The fractionation diagram was found to adequately represent systems where product was lost through denaturation and where more than one purification cut was taken.

Type: Thesis (Doctoral)
Qualification: Ph.D.
Title: Process integration of the initial recovery and purification stages for multiple enzyme production.
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by Proquest
URI: https://discovery.ucl.ac.uk/id/eprint/10109371
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