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Metabolism of Sucrose and Its Five Linkage-isomeric α-D-Glucosyl-D-fructoses by Klebsiella pneumoniae

J. Thompson, S. A. Robrish, S. Immel, F. W. Lichtenthaler, B. G. Hall, and A. Pikis

J. Biol. Chem. 2001, 276, 37415-37425.

Klebsiella pneumoniae is presently unique among bacterial species in its ability to metabolize not only sucrose but also its five linkage-isomeric α-D-glucosyl-D-fructoses: trehalulose, turanose, maltulose, leucrose, and palatinose. Growth on the isomeric compounds induced a protein of molecular mass ~ 50 kDa that was not present in sucrose-grown cells and which we have identified as an NAD+ and metal ion-dependent 6-phospho- α-glucosidase (AglB). The aglB gene has been cloned and sequenced, and AglB (Mr = 49,256) has been purified from a high expression system using the chromogenic p-nitrophenyl α-glucopyranoside 6-phosphate as substrate. Phospho-α-glucosidase catalyzed the hydrolysis of a wide variety of 6-phospho-α-glucosides including maltose-6'-phosphate, maltitol-6-phosphate, isomaltose-6'-phosphate, and all five 6'-phosphorylated isomers of sucrose (Km ~ 1-5 mM) yet did not hydrolyze sucrose-6-phosphate. By contrast, purified sucrose-6-phosphate hydrolase (Mr ~ 53,000) hydrolyzed only sucrose-6-phosphate (Km ~ 80 ÁM). Differences in molecular shape and lipophilicity potential between sucrose and its isomers may be important determinants for substrate discrimination by the two phosphoglucosyl hydrolases. Phospho-α-glucosidase and sucrose-6-phosphate hydrolase exhibit no significant homology, and by sequence-based alignment, the two enzymes are assigned to Families 4 and 32, respectively, of the glycosyl hydrolase superfamily. The phospho-α-glucosidase gene (aglB) lies adjacent to a second gene (aglA), which encodes an EII(CB) component of the phosphoenolpyruvate-dependent sugar:phosphotransferase system. We suggest that the products of the two genes facilitate the phosphorylative translocation and subsequent hydrolysis of the five α-D-glucosyl-D-fructoses by K. pneumoniae.

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