High glycolytic flux improves pyruvate production by a metabolically engineered Escherichia coli strain

dc.contributor.author	Zhu, Y	en
dc.contributor.author	Eiteman, MA	en
dc.contributor.author	Altman, R	en
dc.contributor.author	Altman, E	en
dc.date.accessioned	2014-06-06T06:48:08Z
dc.date.available	2014-06-06T06:48:08Z
dc.date.issued	2008	en
dc.identifier.issn	00992240	en
dc.identifier.uri	http://dx.doi.org/10.1128/AEM.01610-08	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/3979
dc.subject.other	Batch data processing	en
dc.subject.other	Enzymes	en
dc.subject.other	Escherichia coli	en
dc.subject.other	Glucose	en
dc.subject.other	Phosphorus	en
dc.subject.other	Exponential feeding	en
dc.subject.other	Fed-batch	en
dc.subject.other	Formation rates	en
dc.subject.other	Glycolytic fluxes	en
dc.subject.other	Glycolytic rates	en
dc.subject.other	Lactate dehydrogenases	en
dc.subject.other	NADH oxidases	en
dc.subject.other	Nutrient limitations	en
dc.subject.other	Osmoprotectant	en
dc.subject.other	Phosphoenolpyruvate	en
dc.subject.other	Pyruvate	en
dc.subject.other	Pyruvate dehydrogenase complexes	en
dc.subject.other	Pyruvate formate lyase	en
dc.subject.other	Pyruvate oxidases	en
dc.subject.other	Pyruvate productions	en
dc.subject.other	Synthase	en
dc.subject.other	Chemostats	en
dc.subject.other	acetic acid	en
dc.subject.other	formate acetyltransferase	en
dc.subject.other	glucose	en
dc.subject.other	lactate dehydrogenase	en
dc.subject.other	lyase	en
dc.subject.other	nitrogen	en
dc.subject.other	phosphorus	en
dc.subject.other	pyruvate dehydrogenase	en
dc.subject.other	pyruvate oxidase	en
dc.subject.other	pyruvate water dikinase	en
dc.subject.other	pyruvic acid	en
dc.subject.other	reduced nicotinamide adenine dinucleotide dehydrogenase	en
dc.subject.other	synthetase	en
dc.subject.other	unclassified drug	en
dc.subject.other	bioengineering	en
dc.subject.other	chemostat	en
dc.subject.other	coliform bacterium	en
dc.subject.other	enzyme activity	en
dc.subject.other	metabolism	en
dc.subject.other	mutation	en
dc.subject.other	organic acid	en
dc.subject.other	article	en
dc.subject.other	chemostat	en
dc.subject.other	controlled study	en
dc.subject.other	enzyme activity	en
dc.subject.other	enzyme mechanism	en
dc.subject.other	Escherichia coli	en
dc.subject.other	fed batch culture	en
dc.subject.other	gene mutation	en
dc.subject.other	glycolysis	en
dc.subject.other	nonhuman	en
dc.subject.other	protein expression	en
dc.subject.other	Acetic Acid	en
dc.subject.other	Enzymes	en
dc.subject.other	Escherichia coli	en
dc.subject.other	Escherichia coli Proteins	en
dc.subject.other	Glucose	en
dc.subject.other	Glycolysis	en
dc.subject.other	Metabolic Networks and Pathways	en
dc.subject.other	Models, Biological	en
dc.subject.other	Mutation	en
dc.subject.other	Nitrogen	en
dc.subject.other	Phosphorus	en
dc.subject.other	Pyruvic Acid	en
dc.subject.other	Arca	en
dc.subject.other	Escherichia coli	en
dc.title	High glycolytic flux improves pyruvate production by a metabolically engineered Escherichia coli strain	en
heal.type	journalArticle	en
heal.identifier.primary	10.1128/AEM.01610-08	en
heal.publicationDate	2008	en
heal.abstract	We report pyruvate formation in Escherichia coli strain ALS929 containing mutations in the aceEF, pfl, poxB, pps, and ldhA genes which encode, respectively, the pyruvate dehydrogenase complex, pyruvate formate lyase, pyruvate oxidase, phosphoenolpyruvate synthase, and lactate dehydrogenase. The glycolytic rate and pyruvate productivity were compared using glucose-, acetate-, nitrogen-, or phosphorus-limited chemostats at a growth rate of 0.15 h-1. Of these four nutrient limitation conditions, growth under acetate limitation resulted in the highest glycolytic flux (1.60 g/g · h), pyruvate formation rate (1.11 g/g · h), and pyruvate yield (0.70 g/g). Additional mutations in atpFH and arcA (strain ALS1059) further elevated the steady-state glycolytic flux to 2.38 g/g · h in an acetate-limited chemostat, with heterologous NADH oxidase expression causing only modest additional improvement. A fed-batch process with strain ALS1059 using defined medium with 5 mM betaine as osmoprotectant and an exponential feeding rate of 0.15 h-1 achieved 90 g/liter pyruvate, with an overall productivity of 2.1 g/liter · h and yield of 0.68 g/g. Copyright © 2008, American Society for Microbiology. All Rights Reserved.	en
heal.journalName	Applied and Environmental Microbiology	en
dc.identifier.issue	21	en
dc.identifier.volume	74	en
dc.identifier.doi	10.1128/AEM.01610-08	en
dc.identifier.spage	6649	en
dc.identifier.epage	6655	en