1999
DOI: 10.1021/bp990103p
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Biosynthetic Burden and Plasmid Burden Limit Expression of Chromosomally Integrated Heterologous Genes (pdc,adhB) in Escherichiacoli

Abstract: Previous studies have shown an unexpectedly high nutrient requirement for efficient ethanol production by ethanologenic recombinants of Escherichia coli B such as LY01 which contain chromosomally integrated Zymomonas mobilis genes (pdc,adhB) encoding the ethanol pathway. The basis for this requirement has been identified as a media-dependent effect on the expression of the Z. mobilis genes rather than a nutritional limitation. Ethanol production was substantially increased without additional nutrients simply b… Show more

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Cited by 52 publications
(51 citation statements)
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“…Ethanologenic E. coli KO11 was previously constructed by the chromosomal integration of Zymomonas mobilis genes encoding a pyruvate decarboxylase with a low K m (pdc) and alcohol dehydrogenase II (adhB) (37). This strain ferments hexoses and pentoses to ethanol with high efficiency in rich laboratory media (22,29,37). However, volumetric productivity was limited by low cell mass (low biocatalyst concentration) when fermentations were conducted with minerals and less expensive commercial nutrients (29,51,52).…”
mentioning
confidence: 99%
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“…Ethanologenic E. coli KO11 was previously constructed by the chromosomal integration of Zymomonas mobilis genes encoding a pyruvate decarboxylase with a low K m (pdc) and alcohol dehydrogenase II (adhB) (37). This strain ferments hexoses and pentoses to ethanol with high efficiency in rich laboratory media (22,29,37). However, volumetric productivity was limited by low cell mass (low biocatalyst concentration) when fermentations were conducted with minerals and less expensive commercial nutrients (29,51,52).…”
mentioning
confidence: 99%
“…This strain ferments hexoses and pentoses to ethanol with high efficiency in rich laboratory media (22,29,37). However, volumetric productivity was limited by low cell mass (low biocatalyst concentration) when fermentations were conducted with minerals and less expensive commercial nutrients (29,51,52). Similar problems have been reported for the production of other products using genetically engineered strains as biocatalysts (3,9,10,53).…”
mentioning
confidence: 99%
“…Undesirable changes, such as reduced growth, decreased glycolytic flux, and low volumetric productivity, are generally attributed to a lack of ATP (19, 50), creation of futile cycles (10,11,37), changes in intracellular metabolite pools, or a metabolic imbalance (2,7,9,13,28,51,53). Often, these detrimental effects are masked by abundant complex nutrients in laboratory media and are apparent only in more minimal media (7,10,11,31).Salmonella enterica serovar Typhimurium was engineered for succinate production by increasing expression of pyc encoding pyruvate carboxylase (50). Although succinate production increased, the growth rate declined by 18% and the glycolytic flux decreased by 40%.…”
mentioning
confidence: 99%
“…Fermentations were carried out in L broth containing 10% xylose as previously described by using 500-ml vessels (29). The cultures were started with an initial cell concentration of 0.33 g (dry weight) of cells ml Ϫ1 and were incubated for 48 h. Temperature (35°C), pH (pH 6.5), and agitation (100 rpm) were controlled.…”
Section: Methodsmentioning
confidence: 99%
“…Under these conditions, the lower limit for measurement of ethyl acetate in broth was approximately 20 mg liter Ϫ1 . Ethanol was measured as previously described (29). Materials.…”
Section: Methodsmentioning
confidence: 99%