Metabolic flux analysis of Escherichia coli expressing the Bacillus subtilis acetolactate synthase in batch and continuous cultures

Metabolically engineered Escherichia coli expressing the B. subtilis acetolactate synthase has shown to be capable of reducing acetate accumulation. This reduction subsequently led to a significant enhancement in recombinant protein production. The main focus of this study is to systematically exami...

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Bibliographic Details
Published in:Biotechnology and bioengineering 1999-06, Vol.63 (6), p.737-749
Main Authors: Aristidou, Aristos A., San, Ka-Yiu, Bennett, George N.
Format: Article
Language:English
Subjects:
Acetic acid
Acetolactate Synthase - genetics
Acetolactate Synthase - metabolism
Adenosine Triphosphate - metabolism
ALS
Anaerobiosis
Bacillus subtilis
Bacillus subtilis - enzymology
Batch cell culture
Biological and medical sciences
Biotechnology
Biotechnology - methods
Cell Division
Continuous cell culture
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Fermentation
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
metabolic flux analysis
Metabolism
Methods. Procedures. Technologies
Models, Biological
Modification of gene expression level
NAD - metabolism
Proteins
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
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Summary:Metabolically engineered Escherichia coli expressing the B. subtilis acetolactate synthase has shown to be capable of reducing acetate accumulation. This reduction subsequently led to a significant enhancement in recombinant protein production. The main focus of this study is to systematically examine the effect of ALS in the metabolic patterns of E. coli in batch and continuous culture. The specific acetate production rate of a strain carrying the B. subtilis als gene is 75% lower than that of the control strain (host carrying the control plasmid pACYC184) in batch cultures. The ALS strain is further demonstrated to be capable of maintaining a reduced specific acetate production rate in continuous cultures at dilution rates ranging from 0.1 to 0.4 h−1. In addition, this ALS strain is shown to have a higher ATP yield and lower maintenance coefficient. The metabolic flux analysis of carbon flux distribution of the central metabolic pathways and at the pyruvate branch point reveals that this strain has the ability to channel excess pyruvate to the much less toxic compound, acetoin. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 63: 737–749, 1999.
ISSN:0006-3592
1097-0290
DOI:10.1002/(SICI)1097-0290(19990620)63:6<737::AID-BIT12>3.0.CO;2-9