Glycobiology, Vol 9, 31-41, Copyright © 1999 by Oxford University Press
SJ Stasinopoulos, PR Fisher, BA Stone and VA Stanisich
Genes essential for the production of a linear, bacterial (1-->3)-beta-
glucan, curdlan, have been cloned for the first time from Agrobacterium sp.
ATCC31749. The genes occurred in two, nonoverlapping, genomic fragments
that complemented different sets of curdlan( crd )-deficient
transposon-insertion mutations. These were detected as colonies that failed
to stain with aniline blue, a (1-->3)-beta-glucan specific dye. One
fragment carried a biosynthetic gene cluster (locus I) containing the
putative curdlan synthase gene, crdS, and at least two other crd genes. The
second fragment may contain only a single crd gene (locus II).
Determination of the DNA sequence adjacent to several locus I mutations
revealed homology to known sequences only in the cases of crdS mutations.
Complete sequencing of the 1623 bp crdS gene revealed highest similarities
between the predicted CrdS protein (540 amino acids) and glycosyl
transferases with repetitive action patterns. These include bacterial
cellulose synthases (and their homologs), which form
(1-->4)-beta-glucans. No similarity was detected with putative
(1-->3)- beta-glucan synthases from yeasts and filamentous fungi.
Whatever the determinants of the linkage specificity of these beta-glucan
synthases might be, these results raise the possibility that
(1-->3)-beta-glucans and (1-->4)-beta-glucans are formed by related
catalytic polypeptides.
ORIGINAL ARTICLES
Detection of two loci involved in (1-->3)-beta-glucan (curdlan) biosynthesis by Agrobacterium sp. ATCC31749, and comparative sequence analysis of the putative curdlan synthase gene
Department of Biochemistry and Department of Microbiology, La Trobe University, Bundoora 3083, Australia.
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