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Fermentative glycolysis with purified Escherichia coli enzymes for in vitro ATP production and evaluating an engineered enzyme

Stevenson, Bradley; Liu, Jian-Wei; Kuchel, Philip W; Ollis, David


Each of the twelve enzymes for glycolytic fermentation, eleven from Escherichia coli and one from Saccharomyces cerevisiae, have been over-expressed in E. coli and purified with His-tags. Simple assays have been developed for each enzyme and they have been assembled for fermentation of glucose to ethanol. Phosphorus-31 NMR revealed that this in vitro reaction accumulates fructose 1,6-bisphosphate while recycling the cofactors NAD+ and ATP. This reaction represents a defined ATP-regeneration...[Show more]

CollectionsANU Research Publications
Date published: 2012
Type: Journal article
Source: Journal of Biotechnology
DOI: 10.1016/j.jbiotec.2011.09.019


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