Heterologous expression of Escherichia coli ppsA (phosphoenolpyruvate synthetase) and galU (UDP-glucose pyrophosphorylase) genes in Corynebacterium glutamicum, and its impact on trehalose synthesis
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Date
2005
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ACADEMIC PRESS INC ELSEVIER SCIENCE
Abstract
Trehalose is a disaccharide with a wide range of applications in the food industry. We recently proposed a strategy for trehalose production based on a Corynebacterium glutamicum strain expressing the Escherichia coli enzyme UDP-glucose pyrophosphorylase (GalU).
Biochemical network analysis suggest a further bottleneck for trehalose synthesis resulting from the coupling of phosphotransferase (PTS) mediated glucose uptake, and glucose catabolism in C glutamicum. To overcome this coupling, we propose the expression of E coli phosphoenolpyruvate synthetase (PpsA), in addition to GalU expression, in C glutamicum.
Although GalU expression improved trehalose synthesis in C glutamicum, the simultaneous expression of GalU and PpsA did not result in a further increase in trehalose yield, but resulted in an increased catabolic rate of glucose, which could be ascribed to the operation of a futile cycle between phosphoenolpyruvate and pyruvate.
The impact of GalU and PpsA expression on polysaccharide content, side product excretion and metabolic fluxes is discussed, as well as alternative ways to decouple glucose uptake and catabolism, in order to increase trehalose yield. (c) 2005 Elsevier Inc. All rights reserved.
Biochemical network analysis suggest a further bottleneck for trehalose synthesis resulting from the coupling of phosphotransferase (PTS) mediated glucose uptake, and glucose catabolism in C glutamicum. To overcome this coupling, we propose the expression of E coli phosphoenolpyruvate synthetase (PpsA), in addition to GalU expression, in C glutamicum.
Although GalU expression improved trehalose synthesis in C glutamicum, the simultaneous expression of GalU and PpsA did not result in a further increase in trehalose yield, but resulted in an increased catabolic rate of glucose, which could be ascribed to the operation of a futile cycle between phosphoenolpyruvate and pyruvate.
The impact of GalU and PpsA expression on polysaccharide content, side product excretion and metabolic fluxes is discussed, as well as alternative ways to decouple glucose uptake and catabolism, in order to increase trehalose yield. (c) 2005 Elsevier Inc. All rights reserved.
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Keywords
Corynebacterium glutamicum, PpsA, GalU, trehalose, METABOLIC CAPABILITIES, GLYCOGEN-SYNTHESIS, TRANSFORMATION, SYNTHASE, ENZYME, DNA