Document Detail


The effect of thermal deactivation on the properties and processing characteristics of E. coli.
MedLine Citation:
PMID:  8987527     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recombinant microorganisms are often employed to produce proteins of commercial significance. It is important to render such organisms non-viable before culture fluids are released from the fermenter. Thermal deactivation is a superficially attractive option because of its simplicity. The effects of such a thermal deactivation step at the end of a recombinant fermentation are reported in this study. In particular, the consequences of this treatment for down-stream process operations, namely homogenisation and centrifugation, are analysed and discussed. Homogenisation efficiency was adversely affected by a simple treatment whereby cells are raised to 65 degrees C from stationary phase. Cell debris size was also significantly increased. These changes could be partially explained by an increase in fractional peptidoglycan crosslinkage and a decrease in mean cell length. A simple process modification removed these detrimental effects. Specifically, the addition of 15 g of glucose 15 mins prior to thermal deactivation enhances downstream processing. Disruption efficiency is increased above that for stationary-phase cells and the resultant cell debris size is significantly reduced, theoretically aiding inclusion body purification. This novel process modification demonstrates that thermal deactivation may be employed to prevent the release of viable recombinant organisms while providing a broth with desirable processing characteristics. It also emphasises the need to optimise any bioprocess as an interconnected sequence of units.
Authors:
M A Collis; B K O'Neill; C J Thomas; A P Middelberg
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Bioseparation     Volume:  6     ISSN:  0923-179X     ISO Abbreviation:  Bioseparation     Publication Date:  1996 Feb 
Date Detail:
Created Date:  1997-02-13     Completed Date:  1997-02-13     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9011423     Medline TA:  Bioseparation     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  55-63     Citation Subset:  B    
Affiliation:
Department of Chemical Engineering, University of Adelaide, Australia.
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MeSH Terms
Descriptor/Qualifier:
Bioreactors
Biotechnology
Colony Count, Microbial
Escherichia coli / genetics,  growth & development,  metabolism*
Fermentation
Hot Temperature
Molecular Weight
Peptidoglycan / chemistry
Recombinant Proteins / biosynthesis
Safety
Sterilization
Chemical
Reg. No./Substance:
0/Peptidoglycan; 0/Recombinant Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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