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Literature summary extracted from

  • Suginta, W.; Songsiriritthigul, C.; Kobdaj, A.; Opassiri, R.; Svasti, J.
    Mutations of Trp275 and Trp397 altered the binding selectivity of Vibrio carchariae chitinase A (2007), Biochim. Biophys. Acta, 1770, 1151-1160.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.14
-
 Vibrio harveyi

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.14 D392N mutation completely abolishes hydrolyzing activity against colloidal chitin and greatly reduces the hydrolyzing activity against the p-nitrophenyl-N,N'-diacetylchitobiose. Km-value for p-nitrophenyl-N,N'-diacetylchitobiose is 4fold lower than wild-type value, turnover-number is 6.9fold lower than wild-type value Vibrio harveyi
3.2.1.14 W168G mutation completely abolishes hydrolyzing activity against colloidal chitin and greatly reduces the hydrolyzing activity against the p-nitrophenyl-N,N'-diacetylchitobiose. Km-value for p-nitrophenyl-N,N'-diacetylchitobiose is nearly identical to wild-type value, turnover-number is 19.5fold lower than wild-type value Vibrio harveyi
3.2.1.14 W275G Km-value for p-nitrophenyl-N,N'-diacetylchitobiose is 4.6fold lower than wild-type value, turnover-number is 18fold lower than wild-type value Vibrio harveyi
3.2.1.14 W397F mutation enhances hydrolyzing activity towards p-nitrophenyl-N,N'-diacetylchitobiose to 142% and slightly increases activity towards colloidal chitin to 102.5%. Km-value for p-nitrophenyl-N,N'-diacetylchitobiose is 1.16fold higher than wild-type value, turnover-number is 1.2fold higher than wild-type value Vibrio harveyi
3.2.1.14 W570G mutation completely abolishes hydrolyzing activity against colloidal chitin and greatly reduces the hydrolyzing activity against the p-nitrophenyl-N,N'-diacetylchitobiose (5% of wild-type activity) Vibrio harveyi
3.2.1.14 Y171G mutation completely abolishes hydrolyzing activity against colloidal chitin and greatly reduces the hydrolyzing activity against the p-nitrophenyl-N,N'-diacetylchitobiose. Km-value for p-nitrophenyl-N,N'-diacetylchitobiose is 2.5fold lower than wild-type value, turnover-number is 9.7fold lower than wild-type value Vibrio harveyi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.14 0.062
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme W275G Vibrio harveyi
3.2.1.14 0.071
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme D392N Vibrio harveyi
3.2.1.14 0.115
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme Y171G Vibrio harveyi
3.2.1.14 0.278
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme W168G Vibrio harveyi
3.2.1.14 0.288
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, wild-type enzyme Vibrio harveyi
3.2.1.14 0.315
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme W397F Vibrio harveyi

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.14 Vibrio harveyi
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.14
-
 Vibrio harveyi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.14 chitin + H2O
-
 Vibrio harveyi ?
-
 ?
3.2.1.14 N,N',N'',N''',N'''',N'''''-hexaacetylchitohexaose + H2O the time course of hydrolysis exhibits distinct hydrolytic patterns between wild-type and mutants W275G and W397F. Wild-type enzyme initially hydrolyzes N,N',N'',N''',N'''',N'''''-hexaacetylchitohexaose to N,N',N'',N'''-tetraacetylchitotetraose and N,N'-diacetylchitobiose, and finally N,N'-diacetylchitobiose is formed as a major end product. W275G primarily creates N,N'-diacetylchitobiose to N,N',N'',N''',N''''-pentaacetylchitopentaose intermediates, and later N,N'-diacetylchitobiose and N,N',N''-triacetylchitotriose are formed as stable products. W397F initially produces (GlcNAc)1 to N,N',N'',N''',N''''-pentaacetylchitopentaose, and then the N,N',N''-triacetylchitotriose to N,N',N'',N''',N''''-pentaacetylchitopentaose are broken down to (GlcNAc)1 and N,N'-diacetylchitobiose end products Vibrio harveyi ?
-
 ?
3.2.1.14 p-nitrophenyl-N,N'-diacetylchitobiose + H2O
-
 Vibrio harveyi p-nitrophenol + N,N'-diacetylchitobiose
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.14 chitinase A
-
 Vibrio harveyi

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.14 0.11
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme W168G Vibrio harveyi
3.2.1.14 0.12
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme W275G Vibrio harveyi
3.2.1.14 0.21
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme Y171G Vibrio harveyi
3.2.1.14 0.31
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme D392N Vibrio harveyi
3.2.1.14 2.15
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, wild-type enzyme Vibrio harveyi
3.2.1.14 2.59
-
 p-nitrophenyl-N,N'-diacetylchitobiose pH 5.0, 30°C, mutant enzyme W397F Vibrio harveyi