Affiliation:
1. Center for Molecular BioEngineering, Department of Biological and Agricultural Engineering, University of Georgia, Athens, Georgia 30602
Abstract
ABSTRACT
Anaplerotic enzyme reactions are those which replenish tricarboxylic acid intermediates that are withdrawn for the synthesis of biomass. In this study, we examined recombinant protein production in
Escherichia coli
containing activity in an additional anaplerotic enzyme, pyruvate carboxylase. In batch fermentations, the presence of pyruvate carboxylase resulted in 68% greater production of the model protein, β-galactosidase, 41% greater cell yield, and 57% lower acetate concentration. We discuss why these results indicate that acetate concentration does not limit cell growth and protein synthesis, as predicted by other researchers, and suggest instead that the rate of acetate formation represents an inefficient consumption of glucose carbon, which is reduced by the presence of pyruvate carboxylase.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Reference38 articles.
1. Åkesson, M., E. N. Karlsson, P. Hagander, J. P. Axelsson, and A. Tocaj. 1999. On-line detection of acetate formation in Escherichia coli cultures using dissolved oxygen responses to feed transients. Biotechnol. Bioeng.65:590-598.
2. Amann, E., B. Ochs, and K. J. Abel. 1988. Tightly regulated tac vectors useful for the expression of unfused and fused proteins in Escherichia coli.Gene69:301-315.
3. Aristidou, A. A., K. Y. San, and G. N. Bennett. 1994. Modification of central metabolic pathway in Escherichia coli to reduce acetate expression of the Bacillus subtilis acetolactate synthase gene. Biotechnol. Bioeng.44:944-951.
4. Metabolic engineering of Escherichia coli to enhance recombinant protein production through acetate reduction
5. Aristidou, A. A., K. Y. San, and G. N. Bennett. 1999. Improvement of biomass yield and recombinant gene expression in Escherichia coli by using fructose as the primary carbon source. Biotechnol. Prog.15:140-145.
Cited by
52 articles.
订阅此论文施引文献
订阅此论文施引文献，注册后可以免费订阅5篇论文的施引文献，订阅后可以查看论文全部施引文献