Literature summary for 3.2.1.14 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.

Cloned(Commentary)

Cloned (Comment)	Organism
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)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

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

Organism

Organism	UniProt	Comment	Textmining
Nicotiana tabacum	Q43576	-	-

Purification (Commentary)

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

Source Tissue

Source Tissue	Comment	Organism	Textmining
leaf	-	Nicotiana tabacum	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
glycolchitin + H2O	-	Nicotiana tabacum	?	-	?
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	?	-	?
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	-	?
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	-	?
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

Subunits	Comment	Organism
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

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

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.3	-	-	Nicotiana tabacum

pH Stability

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

General Information

General Information	Comment	Organism
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