Document Detail

Characterization of the metabolic burden on Escherichia coli DH1 cells imposed by the presence of a plasmid containing a gene therapy sequence.
MedLine Citation:
PMID:  15532038     Owner:  NLM     Status:  MEDLINE    
The presence of a plasmid, containing gene sequences for DNA immunotherapy that are not expressed in microbial culture, imposed a degradation in bioreactor performance in cultures of the host E. coli strain. Significant decreases in growth rate (24%) and biomass yield (7%) and a corresponding increase in overflow metabolism were observed in a strain containing a therapeutic sequence (a hepatitis B antigen under the control of a CMV promotor). The observed increase in overflow metabolism was incorporated into a Metabolic Flux Analysis (MFA) model (as acetate secretion). Metabolic flux analysis revealed an increase in TCA cycle flux, consistent with an increased respiration rate observed in plasmid-containing cells. These effects are thought to result from increased ATP synthesis requirements (24%) arising from the expression of the Kanr plasmid marker gene whose product accounted for 18% of the cell protein of the plasmid-containing strain. These factors will necessitate significantly higher aeration and agitation rates or lower nutrient feed rates in high-density cultures than would be expected for plasmid-free cultures.
A Rozkov; C A Avignone-Rossa; P F Ertl; P Jones; R D O'Kennedy; J J Smith; J W Dale; M E Bushell
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  88     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2004 Dec 
Date Detail:
Created Date:  2004-11-30     Completed Date:  2005-05-11     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  909-15     Citation Subset:  IM    
Copyright Information:
2004 Wiley Periodicals, Inc.
Microbial Sciences Group, School of Biomedical and Molecular Sciences, University of Surrey, Guildford, Surrey, GU2 7XH, UK.
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MeSH Terms
Base Sequence
Computer Simulation
Escherichia coli / genetics*,  metabolism*
Escherichia coli Proteins / genetics,  metabolism
Gene Expression Regulation, Bacterial / physiology*
Gene Therapy / methods
Hepatitis B Antigens / genetics*
Models, Biological*
Molecular Sequence Data
Plasmids / genetics*,  metabolism*
Reg. No./Substance:
0/Escherichia coli Proteins; 0/Hepatitis B Antigens

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

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