Journal of Bacteriology, July 1999, p. 4176-4184, Vol. 181, No. 14
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
-Glucosidase and a Periplasmic-Binding-Protein-Dependent
Transport System for
-Glucosides
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
Received 4 February 1998/Accepted 29 April 1999
The most abundant carbon source transported into legume root
nodules is photosynthetically produced sucrose, yet the importance of
its metabolism by rhizobia in planta is not yet known. To identify genes involved in sucrose uptake and hydrolysis, we screened a Sinorhizobium meliloti genomic library and discovered a
segment of S. meliloti DNA which allows Ralstonia
eutropha to grow on the
-glucosides sucrose, maltose, and
trehalose. Tn5 mutagenesis localized the required genes to
a 6.8-kb region containing five open reading frames which were named
agl, for
-glucoside utilization. Four of these
(aglE, aglF, aglG, and
aglK) appear to encode a periplasmic-binding-protein-dependent sugar transport system, and one
(aglA) appears to encode an
-glucosidase with homology to family 13 of glycosyl hydrolases. Cosmid-borne agl genes
permit uptake of radiolabeled sucrose into R. eutropha
cells. Analysis of the properties of agl mutants suggests
that S. meliloti possesses at least one additional
-glucosidase as well as a lower-affinity transport system for
-glucosides. It is possible that the Fix+ phenotype of
agl mutants on alfalfa is due to these additional functions. Loci found by DNA sequencing to be adjacent to
aglEFGAK include a probable regulatory gene
(aglR), zwf and edd, which encode
the first two enzymes of the Entner-Doudoroff pathway, pgl,
which shows homology to a gene encoding a putative
phosphogluconolactonase, and a novel Rhizobium-specific
repeat element.
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