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Background  A metabolic regulation study was performed, based upon measurements of enzymatic activities, fermentation performance, and RT-PCR analysis of pathways related to central carbon metabolism, in an ethanologenic  Escherichia coli  strain (CCE14) derived from lineage C. In comparison with previous engineered strains, this  E coli  derivative has a higher ethanol production rate in mineral medium, as a result of the elevated heterologous expression of the chromosomally integrated genes encoding PDC  Zm   and ADH  Zm   (pyruvate decarboxylase and alcohol dehydrogenase from  Zymomonas mobilis ). It is suggested that this behavior might be due to lineage differences between  E. coli  W and C.    Results  This study demonstrated that the glycolytic flux is controlled, in this case, by reactions outside glycolysis,  i.e. , the fermentative pathways. Changes in ethanol production rate in this ethanologenic strain result in low organic acid production rates, and high glycolytic and ethanologenic fluxes, that correlate with enhanced transcription and enzymatic activity levels of PDC  Zm   and ADH  Zm  . Furthermore, a higher ethanol yield (90% of the theoretical) in glucose-mineral media was obtained with CCE14 in comparison with previous engineered  E. coli  strains, such as KO11, that produces a 70% yield under the same conditions.    Conclusion  Results suggest that a higher ethanol formation rate, caused by ahigher PDC  Zm   and ADH  Zm   activities induces a metabolic state that cells compensate through enhanced glucose transport, ATP synthesis, and NAD-NADH+H turnover rates. These results show that glycolytic enzymatic activities, present in  E. coli  W and C under fermentative conditions, are sufficient to contend with increases in glucose consumption and product formation rates.

Metabolic regulation analysis of an ethanologenic Escherichia coli strain based on RT-PCR and enzymatic activities