Adenosine diphosphate sugar pyrophosphatase prevents glycogen biosynthesis in Escherichia coli.
An adenosine diphosphate sugar pyrophosphatase (ASPPase, EC ) has been characterized by using Escherichia coli. This enzyme, whose activities in the cell are inversely correlated with the intracellular glycogen content and the glucose concentration in the culture medium, hydrolyzes ADP-glucose, the precursor molecule of glycogen biosynthesis. ASPPase was purified to apparent homogeneity (over 3,000-fold), and sequence analyses revealed that it is a member of the ubiquitously distributed group of nucleotide pyrophosphatases designated as "nudix" hydrolases. Insertional mutagenesis experiments leading to the inactivation of the ASPPase encoding gene, aspP, produced cells with marginally low enzymatic activities and higher glycogen content than wild-type bacteria. aspP was cloned into an expression vector and introduced into E. coli. Transformed cells were shown to contain a dramatically reduced amount of glycogen, as compared with the untransformed bacteria. No pleiotropic changes in the bacterial growth occurred in both the aspP-overexpressing and aspP-deficient strains. The overall results pinpoint the reaction catalyzed by ASPPase as a potential step of regulating glycogen biosynthesis in E. coli.
Moreno-Bruna
B
B
Instituto de Agrobiotecnologia y Recursos Naturales, Universidad Pública de Navarra/Consejo Superior de Investigaciones Cientificas, Ctra. de Mutilva s/n, Mutilva Baja 31192 Navarra, Spain.
Baroja-Fernández
E
E
Muñoz
F J
FJ
Bastarrica-Berasategui
A
A
Zandueta-Criado
A
A
Rodriguez-López
M
M
Lasa
I
I
Akazawa
T
T
Pozueta-Romero
J
J
D000244:Adenosine Diphosphate / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D006003:Glycogen / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D011755:Pyrophosphatases / Q000378:metabolism*
D000244:Adenosine Diphosphate; D006003:Glycogen; D011755:Pyrophosphatases