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Literature summary for 3.2.1.14 extracted from

  • Hurtado-Guerrero, R.; van Aalten, D.M.
    Structure of Saccharomyces cerevisiae chitinase 1 and screening-based discovery of potent inhibitors (2007), Chem. Biol., 14, 589-599.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of the catalytic domain of ScCTS1 (comprising of amino acids 22 to 315) as a secreted N-terminally His-tagged fusion protein in Pichia pastoris Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment) Organism
crystallization of the native enzyme and the enzyme in complex with 8-chlorotheophylline, kinetin or acetazolamide, sitting drop method, 1.6 A resolution Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
A283S Km-value for mutant enzyme for 4-methylumbelliferyl-N,N',N''-triacetylchitotriose is 0.8 mM versus 1 mM for wild-type enzyme and turnover number is 2.3 per sec versus 1.3 for wild-type enzyme. Inhibition by allosamidin, acetazolamide, and 8-chlorotheophylline are not significantly affected by the mutation, inhibition by kinetin is not detectable Saccharomyces cerevisiae
F210W/A283S similar Km (1.1 versus 1 mM for wild-type) and turnover number kcat (0.8 versus 1.3 for wild-type) as for the wild-type enzyme and the single A283S mutant. Inhibition by allosamidin, acetazolamide, and 8-chlorotheophylline is not significantly affected by the mutation, inhibition by kinetin is not detectable Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
8-chlorotheophylline
-
Saccharomyces cerevisiae
acetazolamide competitive Saccharomyces cerevisiae
allosamidin
-
Saccharomyces cerevisiae
kinetin competitive Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.8
-
4-methylumbelliferyl-N,N',N''-triacetylchitotriose mutant enzyme A283S Saccharomyces cerevisiae
1
-
4-methylumbelliferyl-N,N',N''-triacetylchitotriose wild-type enzyme Saccharomyces cerevisiae
1.1
-
4-methylumbelliferyl-N,N',N''-triacetylchitotriose mutant enzyme F210W/A283S Saccharomyces cerevisiae

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae P29029
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl-N,N',N''-triacetylchitotriose + H2O
-
Saccharomyces cerevisiae ?
-
?

Synonyms

Synonyms Comment Organism
chitinase 1
-
Saccharomyces cerevisiae
CTS1
-
Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.8
-
4-methylumbelliferyl-N,N',N''-triacetylchitotriose mutant enzyme F210W/A283S Saccharomyces cerevisiae
1.3
-
4-methylumbelliferyl-N,N',N''-triacetylchitotriose wild-type enzyme Saccharomyces cerevisiae
2.3
-
4-methylumbelliferyl-N,N',N''-triacetylchitotriose mutant enzyme A283S Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
2.6
-
substrate: 4-methylumbelliferyl-N,N',N''-triacetylchitotriose Saccharomyces cerevisiae

pH Range

pH Minimum pH Maximum Comment Organism
2 4 pH 2: about 80% of maximal activity, pH 4.0: about 55% of maximal activity, substrate: 4-methylumbelliferyl-N,N',N''-triacetylchitotriose Saccharomyces cerevisiae

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.00061
-
allosamidin wild-type enzyme Saccharomyces cerevisiae
0.00066
-
allosamidin mutant enzyme A283S Saccharomyces cerevisiae
0.00146
-
allosamidin mutant enzyme F210W/A283S Saccharomyces cerevisiae
0.0032
-
kinetin wild-type enzyme Saccharomyces cerevisiae
0.021
-
acetazolamide wild-type enzyme Saccharomyces cerevisiae
0.071
-
acetazolamide mutant enzyme A283S Saccharomyces cerevisiae
0.128
-
acetazolamide mutant enzyme F210W/A283S Saccharomyces cerevisiae
0.6
-
8-chlorotheophylline wild-type enzyme Saccharomyces cerevisiae
1.155
-
8-chlorotheophylline mutant enzyme A283S Saccharomyces cerevisiae
1.192
-
8-chlorotheophilline mutant enzyme F210W/A283S Saccharomyces cerevisiae