Abstract
Inner membranes of Bradyrhizobium japonicum strain USDA 110 produced in vitro soluble and insoluble β-(1–3),β-(1–6) glucansFootnote 1. The reaction proceeded through a 90 kDa inner membrane intermediate protein; used UDP-glucose as sugar donor and required Mg2+. Gel chromatography of soluble glucans resolved a cyclic β-(1–3) glucan with a degree of polymerization of eleven from a family of β-(1–3),β-(1–6) glucans with variable degree of polymerization higher than eleven. Bradyrhizobium strains BR4406 and BR8404 isolated from tree legume nodules in Southeast Brazil produce β-(1–3),β-(1–6) glucans very similar to that of B. japonicum. A 100 kDa protein was identified in these strains as intermediates in the synthesis of these glucans. Inner membranes of B. japonicum USDA110, B. japonicum I17, and Bradyrhizobium strains BR4406 and BR8404 incubated with UDP-glucose were unable to synthesize β-(1–2) glucan and lacked the 235 kDa intermediate protein known to be involved in the synthesis of β-(1–2) glucan in Agrobacterium tumefaciens, Rhizobium meliloti and Rhizobium loti.
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Notes
A preliminary account of this work was presented at the 5th International Symposium on the Molecular Genetics of Plant-Microbe Interactions, Interlaken, Switzerland, September 1990
Abbreviations
- EPS=:
-
exopolysaccharides
- CPS=:
-
capsular polysaccharides
- LPS=:
-
lipopolysaccharides
- AMA=:
-
Yeast extract-mannitol medium
- TY=:
-
tryptone-yeast extract
- PMSF=:
-
phenyl methyl sulfonil fluoride
References
Bourne EJ, Huston DH, Weigel H (1961) Complexes between molibdate and acyclic polyhydroxy-compounds. J Chem Soc 35–38
Cohen JL, Miller KJ (1991) A novel membrane-bound glucosyl-transferase from Bradyrhizobium japonicum. J Bacteriol 173: 4271–4276
Chakravorty AK, Zurkowski W, Shine I, Rolfe BG (1982) Symbiotic nitrogen fixation: molecular cloning of Rhizobium genes involved in expolysaccharide and effective nodulation. J Mol App Genet 1: 585–596
Dylan T, Ielpi L, Stanfield S, Kashyap L, Douglas C, Yanofsky M, Nester EW, Helinski DR, Ditta G (1986) Rhizobium meliloti genes required for nodule development are related to chromosomal virulence genes in Agrobacterium tumefaciens chvA. Proc Natl Acad Sci USA 83: 4403–4407
Flowers HM, Batra KK, Kemp J, Hassid WZ (1968) Biosynthesis of insoluble glucans from uridine-diphosphate-d-glucose with enzyme preparations from Phaseolus aureus and Lupinus albus. Plant Physiol 43: 1703–1709
Geremia RA, Cavagniac S, Zorreguieta A, Toro N, Olivares J, Ugalde RA (1987) A Rhizobium meliloti mutant that forms ineffective pseudonodules in alfalfa produces exopolysaccharides but fails to form β-(1–2) glucan. J Bacteriol 169: 880–884
Hassid WZ (1969) Biosynthesis of oligosacharides and polysaccharides in plants. Mechanisms of enzymatic synthesis of complex plant carbohydrates are reviewed. Science 165: 137–144
Iñón de Iannino N, Ugalde RA (1989) Biochemical characterization of avirulent Agrobacterium tumefaciens chvA mutants synthesis and excretion of β-(1–2) glucan. J Bacteriol 171: 2842–2849
Jeanes A, Wise CS, Dimler RJ (1951) Improved techniques in paper chromatography of carbohydrates. Anal Chem 23: 415–420
Leigh J, Signer ER, Walker GC (1985) Exopolysaccharide-deficient mutants of Rhizobium meliloti that form ineffective nodules. Proc Natl Acad Sci USA 82: 6231–6235
Marechal LR, Goldemberg SH (1964) Uridine diphosphate glucose β-1,3-glucan β-3-glucosyltransferase from Euglena gracilis. J Biol Chem 239: 3163–3167
Miller KJ, Gore RS, Johnson R, Benesi AL, Reinhold VN (1990) Cell-associated oligosaccharides of Bradyrhizobium spp. J Bacteriol 172: 136–142
Miller KJ, Kennedy EP, Reinhold VN (1986) Osmotic adaptation by Gram-negative bacteria: possible role for periplasmic oligosaccharides. Science 231: 48–51
Osborn MJ, Munson R (1984) Separation of inner (cytoplasmic) and outer membranes of gram negative bacteria. Methods Enzymol 31A: 642–653
Peat S, Whelan WJ, Lawley HG (1958) The structure of laminarin. The main polymeric linkage. J Chem Soc 724–737
Puvanesarajah V, Schell FM, Stacey G, Douglas CJ, Nester EW (1985) Role for 2-linked-β-d-glucan in the virulence of Agrobacterium tumefaciens. J Bacteriol 164: 102–106
Shematek EM, Braatz JA, Cabib E (1980) Biosynthesis of the yeast cell wall. Preparation and properties of β-(1–3) glucan synthethase. J Biol Chem 255: 888–894
Stanfield SW, Ielpi L, O'Brochta D, Helinski DR, Ditta GS (1988) The ndvA gene product of Rhizobium meliloti is required for β-(1–2) glucan production and has homology to the ATP-binding export protein HlyB. J Bacteriol 170: 3523–3530
Trevelyan WE, Procter DP, Harrison JS (1950) Detection of sugar on paper chromatography. Nature 166: 444–445
Tomos AD, Northcote DH (1978) A protein-glucan intermediate during paramylon synthesis. Biochemistry 174: 283–290
Tung KK, Nordin JH (1968) Structure of the tetrasaccharide produced by the hydrolysis of nigeran by the enzyme mycodextranase. Biochim Biophys Acta 158: 154–156
Williams MVH, Hollingsworth RI, Klein S, Signer ER (1990) The symbiotic defect of Rhizobium meliloti expolysaccharide mutants is supressed by IpsZ +, a gene involved in lipopolysaccharide Biosynthesis. J Biol Chem 172: 2622–2632
Zorreguieta A, Ugalde RA (1986) Formation in Rhizobium and Agrobacterium spp. of a 235 kDa protein intermediate in β-d-(1–2) glucan synthesis. J Bacteriol 167: 947–951
Zorreguieta A, Geremia RA, Cavagniac S, Cangelosi G, Nester EW, Ugalde RA (1988) Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. Plant Microbe Interaction 3: 121–127
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Iñón de Iannino, N., Ugalde, R.A. Biosynthesis of cyclic β-(1–3),β-(1–6) glucan in Bradyrhizobium spp.. Arch. Microbiol. 159, 30–38 (1993). https://doi.org/10.1007/BF00244260
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DOI: https://doi.org/10.1007/BF00244260