Journal of Chemical Technology and Biotechnology, 1999, 73 (3), 208-212
Direct process
integration of cell disruption and fluidised bed adsorption for the recovery
of intracellular proteins
Horst Bierau, Zhanren
Zhang & Andrew Lyddiatt*
Biochemical Recovery Group, Centre for Bioprocess Engineering, School
of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham,
B15 2TT, UK
Tel./Fax:
0121 414 5278, email: A.Lyddiatt@bham.ac.uk (www.bham.ac.uk/BRG)
Key words: cell disruption, bead mill, fluidised bed, protein recovery,
process integration, intracellular proteins
Abstract
An integrated
process for the primary capture of an intracellular enzyme, where cell disruption
is directly coupled with fluidised bed adsorption of the product, was proposed
as a generic approach to benefit the yield and molecular integrity of labile
protein products. The purification of glyceraldehyde 3-phophate dehydrogenase
(G3PDH) from waste brewers' yeast was selected for the demonstration of this
principle. Cell disruption by bead milling was combined with direct adsorption
of the enzyme on a Cibacron Blue derivative of a kieselguhr-agarose composite
adsorbent in a fluidised bed contactor operated immediately downstream of
the cell disrupter. The short process time and immediate sequestration of
product from the hostile disruptate environment facilitated the recovery of
partially purified preparations of this labile enzyme from yeast aged at -20
degrees C for 9 months. The recovered specific activity of 7.6 IU mg(-1) bettered
that expected from the extended time-scale of sequential batch operations
of milling, centrifugation or microfiltration, and fixed bed chromatography.
The potential role for this novel approach to process integration is discussed
in the context of the adsorbent and contactor optimisation necessary to establish
efficient, continuous primary recovery of labile intracellular proteins. (C)
1999 Society of Chemical Industry.