Impact of heterologous expression of Escherichia coli UDP-glucose pyrophosphorylase on trehalose and glycogen synthesis in Corynebacterium glutamicum.
Trehalose is a disaccharide with a wide range of applications in the food industry. We recently proposed a strategy for trehalose production based on improved strains of the gram-positive bacterium Corynebacterium glutamicum. This microorganism synthesizes trehalose through two major pathways, OtsBA and TreYZ, by using UDP-glucose and ADP-glucose, respectively, as the glucosyl donors. In this paper we describe improvement of the UDP-glucose supply through heterologous expression in C. glutamicum of the UDP-glucose pyrophosphorylase gene from Escherichia coli, either expressed alone or coexpressed with the E. coli ots genes (galU otsBA synthetic operon). The impact of such expression on trehalose accumulation and excretion, glycogen accumulation, and the growth pattern of new recombinant strains is described. Expression of the galU otsBA synthetic operon resulted in a sixfold increase in the accumulated and excreted trehalose relative to that in a wild-type strain. Surprisingly, single expression of galU also resulted in an increase in the accumulated trehalose. This increase in trehalose synthesis was abolished upon deletion of the TreYZ pathway. These results proved that UDP-glucose has an important role not only in the OtsBA pathway but also in the TreYZ pathway.
Padilla
Leandro
L
Departmento de Ingeniería y Bioprocesos, Escuela de Ingeniería, Pontificia Universidad Católica de Chile, Casilla 306 Correo 22, Santiago, Chile.
Morbach
Susanne
S
Krämer
Reinhard
R
Agosin
Eduardo
E
D003352:Corynebacterium / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000502:physiology; D005964:Glucosyltransferases / Q000502:physiology; D006003:Glycogen / Q000096:biosynthesis*; D009876:Operon; D010744:Phosphoric Monoester Hydrolases / Q000502:physiology; D014199:Trehalose / Q000096:biosynthesis*; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000235:genetics* / Q000502:physiology; D014532:Uridine Diphosphate Glucose / Q000378:metabolism
D029968:Escherichia coli Proteins; D006003:Glycogen; D014199:Trehalose; D005964:Glucosyltransferases; C023180:trehalose-6-phosphate synthase; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; C121039:galU protein, E coli; C023178:trehalose-phosphatase; D010744:Phosphoric Monoester Hydrolases; D014532:Uridine Diphosphate Glucose