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.
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.
Stasinopoulos
S J
SJ
Department of Biochemistry and Department of Microbiology, La Trobe University, Bundoora 3083, Australia.
Fisher
P R
PR
Stone
B A
BA
Stanisich
V A
VA
D000595:Amino Acid Sequence; D002236:Carbohydrate Conformation; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005936:Glucans / Q000096:biosynthesis* / Q000235:genetics; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics*; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012231:Rhizobium / Q000201:enzymology / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins; D016415:Sequence Alignment; D013379:Substrate Specificity; D047071:beta-Glucans
D005936:Glucans; D008565:Membrane Proteins; D029702:Schizosaccharomyces pombe Proteins; D047071:beta-Glucans; C038459:curdlan; D005964:Glucosyltransferases; C020570:1,3-beta-glucan synthase; D004262:DNA Restriction Enzymes