Modification of central metabolic pathway in escherichia coli to reduce acetate accumulation by heterologous expression of the bacillus subtilis acetolactate synthase gene

A novel metabolic engineering technique involving the redirection ofcellular carbon fluxes was employed to reduce acetate production in an Escherichia coli culture. Metabolic engineering was achieved by cloning E. coli the gene for the Bacillus subtilis acetolactate synthase (ALS), an enzyme capable...

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Bibliographic Details
Published in:Biotechnology and bioengineering 1994-10, Vol.44 (8), p.944-951
Main Authors: Aristidou, Aristos A., San, Ka-Yiu, Bennett, George N.
Format: Article
Language:English
Subjects:
acetate reduction
Bacillus subtilis
Biological and medical sciences
Biotechnology
cloning
Escherichia coli
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Methods. Procedures. Technologies
Modification of gene expression level
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Summary:A novel metabolic engineering technique involving the redirection ofcellular carbon fluxes was employed to reduce acetate production in an Escherichia coli culture. Metabolic engineering was achieved by cloning E. coli the gene for the Bacillus subtilis acetolactate synthase (ALS), an enzyme capable of catalyzing the conversion of pyruvate to nonacidic and less harmful species. The heterologous expression of the ALS catabolic enzyme in Escherichia coli drastically modified the cellular glycolytic fluxes. In particular, acetate excretion, which is a common characteristic of E. coli, as well as a physiological burden, was minimized. The residual acetate level was kept under control and maintained at a level that was below the toxic threshold. The expression of the biologically active ALS enzyme in E. coli did not result in any detectable changes on either cell growth rate or cell yields. The alternative product, acetoin, was shown to be 50 times less harmful than acetate. Similarities in the growth pattern of two different E. coli strains, RR1 and GJT001, under all cultivation conditions suggested that the ability of ALS to reduce acetate accumulation is generic and not strain‐specific. © 1994 John Wiley & Sons, Inc.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.260440810