Rhizobium NodB protein involved in nodulation signal synthesis is a chitooligosaccharide deacetylase.

John, Michael; Röhrig, Horst; Schmidt, Jürgen; Wieneke, Ursula; Schell, Jeff

doi:10.1073/pnas.90.2.625

Cited by 228 publications

(192 citation statements)

References 27 publications

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“…These results indicate that NodB acts before, and independently of, NodA. This is in accord with the results of John et al (1993), who found that nodB encodes an acetylase that presumably removes the acetyl group that initially occupies the position the acyl group will ultimately occupy, and with those of Rohrig et al (1994), who demonstrated that NodA is involved in acylation. Indeed, Atkinson et al (1994) found that when a deacetylated substrate is provided, NodA is able to add the fatty acyl group.…”

Section: Genetic Approaches To Plant Biochemistrysupporting

confidence: 81%

ASPP Meeting: Germinating Ideas in Portland

Chasan¹

1995

Plant Cell

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Section: Genetic Approaches To Plant Biochemistrysupporting

confidence: 81%

ASPP Meeting: Germinating Ideas in Portland

Chasan¹

1995

Plant Cell

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“…The experimental conditions concerning incubation of the oligomers with the exo-glycosidases as well as their subsequent analysis by HPLC are described in the Experimental procedures section. The peaks 1 H , 4 H , 1, 2, 4 correspond to GlcN (GlcN) 4 , GlcNAc (GlcNAc) 2 and (GlcNAc) 4 , respectively, as deduced from their elution times. The peaks a and b correspond to (GlcN) 2 GlcNAcGlcNAc and (GlcN) 3 GlcNAc, respectively.…”

Section: Separation Of Pmp-labelled Chitin and Chitosan Oligomersmentioning

confidence: 99%

“…Elucidation of reaction products using the exo-splitting system. 60 mg of (GlcNAc) 4 was incubated with 1 mU of chitin deacetylase from M. rouxii in a 150-mL mtotal volume of 50 mm glutamate buffer pH 4.5 at 50 8C. After 48 h of incubation the reaction was terminated by heating at 100 8C for 5 min and the reaction mixture was divided in three equal parts.…”

Section: Separation Of Pmp-labelled Chitin and Chitosan Oligomersmentioning

confidence: 99%

“…b-NGlcNAcase activity was assayed using (GlcNAc) 4 as substrate. The reaction mixture (250 mL total volume), consisting of 0.5 mm (GlcNAc) 4 in a 100 mm acetate buffer pH 5.0, was incubated for 15 min at 37 8C, and the amount of GlcNAc produced was measured according to Elson and Morgan [25], with GlcNAc as control. One unit of b-N-GlcNAcase activity is defined as the amount of enzyme which released one mmol of GlcNAc per min.…”

Section: Enzyme Assaysmentioning

confidence: 99%

“…Protein sequence comparisons revealed significant similarities between the fungal chitin deacetylase and the rhizobial nodB proteins, suggesting functional homology of these evolutionary distinct proteins. The functional assignment of the nodB protein in Nod factor biosynthesis, as deduced from its sequence similarity, was subsequently verified biochemically [4]. The purification and characterization of chitin deacetylases from the fungi Colletotrichum lindemuthianum [5±7], Absidia coerulea [8], and Aspergillus nidulans [9] have also been recently reported.…”

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Mode of action of chitin deacetylase from Mucor rouxii on N‐acetylchitooligosaccharides

Tsigos

Zydowicz

Martinou

et al. 1999

European Journal of Biochemistry

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The mode of action of chitin deacetylase from the fungus Mucor rouxii on N-acetylchitooligosaccharides with a degree of polymerization 1±7 has been elucidated.Identification of the sequence of chitin oligomers following enzymatic deacetylation was verified by the alternative use of two specific exo-glycosidases in conjunction with HPLC. The results were further verified by 1 H-NMR spectroscopy.It was observed that the length of the oligomer is important for enzyme action. The enzyme cannot effectively deacetylate chitin oligomers with a degree of polymerization lower than three. Tetra-N-acetylchitotetraose and penta-N-acetylchitopentaose are fully deacetylated by the enzyme, while in the case of tri-N-acetylchitotriose, hexa-N-acetylchitohexaose and hepta-N-acetylchitoheptaose the reducing-end residue always remains intact. Furthermore, the enzyme initially removes an acetyl group from the nonreducing-end residue of all chitin oligomers with a degree of polymerization higher than 2, and further catalyses the hydrolysis of the following acetamido groups in a processive fashion.The results are in agreement with the mode of action that the same enzyme exhibits on partially deacetylated water soluble chitosan polymers.Keywords: chitin deacetylase; chitin oligomers; chitosan oligomers; enzymatic deacetylation.Chitin deacetylase, the enzyme that catalyses the conversion of chitin to chitosan by the deacetylation of N-acetyl-d-glucosamine residues, was initially purified to homogeneity and further characterized from mycelial extracts of the fungus Mucor rouxii [1,2]. Furthermore, a cDNA of the M. rouxii encoding chitin deacetylase was isolated, sequenced and further characterized [3]. Protein sequence comparisons revealed significant similarities between the fungal chitin deacetylase and the rhizobial nodB proteins, suggesting functional homology of these evolutionary distinct proteins. The functional assignment of the nodB protein in Nod factor biosynthesis, as deduced from its sequence similarity, was subsequently verified biochemically [4]. The purification and characterization of chitin deacetylases from the fungi Colletotrichum lindemuthianum [5±7], Absidia coerulea [8], and Aspergillus nidulans [9] have also been recently reported. All enzymes are glycoproteins with molecular masses ranging from 27 to 150 kDa. In all cases the optimum temperature for enzyme activity is 50 8C while the optimum pH varies from 4.5 to 8.5. All enzymes are active on chitin oligomers, while they are not effective in deacetylating waterinsoluble chitin substrates.The mode of action of chitin deacetylase from M. rouxii on partially N-acetylated chitosans has also been recently reported [10]. Using 1 H-NMR and 13 C-NMR spectroscopy it was observed that the enzyme exhibits a`multiple attack' [11] reaction mechanism. According to this mechanism, binding of the enzyme on a chitin chain is followed by a number of sequential deacetylations after which the enzyme binds to another chain.Chitin oligomers, in contrast with their corresponding poly...

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Metal‐Dependent Polysaccharide Deacetylase PgaB

Little

Bamford

Nitz

et al. 2014

Encyclopedia of Inorganic and Bioinorganic Chemistry

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PgaB is a two‐domain protein responsible for the de‐ N ‐acetylation of poly(β‐1,6‐ N ‐acetyl‐ d ‐glucosamine) ( PNAG ). Many bacteria use deacetylated PNAG ( dPNAG ) as a key extracellular matrix component. De‐ N ‐acetylation is a required modification for polymer export and biofilm formation in Escherichia coli . The N‐terminal domain adopts a (β/α) 7 fold common to the CAZy family‐4 carbohydrate esterases, and exhibits low levels of de‐ N ‐acetylation activity on PNAG oligomers. The de‐ N ‐acetylase active site contains an Asp‐His‐His metal coordination site that can bind various divalent metal cations but has optimal activity with Co 2+ , Ni 2+ , and Fe 2+ . The C‐terminal domain of PgaB adopts a (β/α) 8 fold that is structurally similar to glycoside hydrolases. This domain is proposed to bind PNAG as attempts to display hydrolytic activity have been unsuccessful, but the domain is required for de‐ N ‐acetylation in vivo. Here, we summarize recent advances in the functional characterization of PgaB: the structure, metal specificity, and progress toward inhibitor design.

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Rhizobium NodB protein involved in nodulation signal synthesis is a chitooligosaccharide deacetylase.

Cited by 228 publications

References 27 publications

ASPP Meeting: Germinating Ideas in Portland

ASPP Meeting: Germinating Ideas in Portland

Mode of action of chitin deacetylase from Mucor rouxii on N‐acetylchitooligosaccharides

Metal‐Dependent Polysaccharide Deacetylase PgaB

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