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Literature summary extracted from

  • Ohnuma, T.; Numata, T.; Osawa, T.; Mizuhara, M.; Varum, K.M.; Fukamizo, T.
    Crystal structure and mode of action of a class V chitinase from Nicotiana tabacum (2011), Plant Mol. Biol., 75, 291-304.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.14 gene chiV, DNA and amino acid sequence determination and analysis, wild-type and mutant NtChiVs recombinant expression in Escherichia coli strain Tuner(DE3) pLacI. Expression of SeMet-labeled NtChiV in the methionine auxotroph Escherichia coli strain B834(DE3) pLacI Nicotiana tabacum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.14 purified recombinant native and SeMet-labeled wild-type enzymes and mutant E115Q enzyme, hanging drop vapor diffusion method, mixing of 0.001 ml 5 mg/ml protein in water with 0.001 ml of reservoir solution containing 100 mM Tris-HCl, pH 8.5, and 1.7-2.2 M ammonium dihydrogen phosphate, 20°C, 1 months, X-ray diffraction structure determination and analysis at 1.2, 1.6, and 1.8 A resolution, respectively, single-wavelength anomalous dispersion method Nicotiana tabacum

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.14 E115Q site-directed mutagenesis, a catalytically inactive mutant, crystal structure determination and analysis Nicotiana tabacum

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.14 Nicotiana tabacum Q43576
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.14 recombinant wild-type and mutant NtChiVs from Escherichia coli strain Tuner(DE3) pLacI to homogeneity Nicotiana tabacum

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.2.1.14 leaf
-
 Nicotiana tabacum
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.14 glycolchitin + H2O
-
 Nicotiana tabacum ?
-
 ?
3.2.1.14 additional information the enzyme hydrolyzes N-acetylglucosamine oligosaccharides, predominantly the second glycosidic linkage from the non-reducing end. And NtChiV hydrolyzes partially N-acetylated chitosan non-processively. NtChiV does catalyze a transglycosylation reaction, but the extent is negligible over the hydrolysis reaction. Substrate recognition and mode of action, overview Nicotiana tabacum ?
-
 ?
3.2.1.14 N,N',N'',N''', N'''',N'''''-hexaacetylchitohexaose + H2O the enzyme hydrolyzes predominantly the second glycosidic linkage from the non-reducing end Nicotiana tabacum N,N'-diacetylchitobiose + N,N',N'',N'''-tetraacetylchitotetraose
-
 ?
3.2.1.14 N,N',N'',N''',N''''-pentaacetylchitopentaose + H2O the enzyme hydrolyzes predominantly the second glycosidic linkage from the non-reducing end Nicotiana tabacum N,N'-diacetylchitobiose + N,N',N''-triacetylchitotriose
-
 ?
3.2.1.14 N,N',N'',N'''-tetraacetylchitotetraose + H2O the enzyme hydrolyzes predominantly the second glycosidic linkage from the non-reducing end Nicotiana tabacum 2 N,N'-diacetylchitobiose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.14 More enzyme NtChiV adopts a classical (beta/alpha)8-barrel fold, residues 1-233 and 303-348, with an insertion of a small, alpha + beta, domain, residues 234-302. The chitin-binding domain is present in the C-terminus Nicotiana tabacum

Synonyms

EC Number Synonyms Comment Organism
3.2.1.14 class V chitinase
-
 Nicotiana tabacum
3.2.1.14 More the enzyme is a family GH-18 chitinase Nicotiana tabacum
3.2.1.14 NtChiV
-
 Nicotiana tabacum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.14 65
-
-
 Nicotiana tabacum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.14 4 60 purified recombinant enzyme, stable Nicotiana tabacum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.14 6.3
-
-
 Nicotiana tabacum

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.14 4.2 10 purified recombinant enzyme, stable Nicotiana tabacum

General Information

EC Number General Information Comment Organism
3.2.1.14 additional information NtChiV is a non-processive enzyme. The aglycon binding site of NtChiV appears to consist of three subsites, +1, +2, and +3. Structural comparison with other family GH-18 chitinases, overview Nicotiana tabacum