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 (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 | 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 | Comment | Organism | Structure |
---|---|---|---|
8-chlorotheophylline | - |
Saccharomyces cerevisiae | |
acetazolamide | competitive | Saccharomyces cerevisiae | |
allosamidin | - |
Saccharomyces cerevisiae | |
kinetin | competitive | Saccharomyces cerevisiae |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P29029 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-methylumbelliferyl-N,N',N''-triacetylchitotriose + H2O | - |
Saccharomyces cerevisiae | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
chitinase 1 | - |
Saccharomyces cerevisiae |
CTS1 | - |
Saccharomyces cerevisiae |
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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
2.6 | - |
substrate: 4-methylumbelliferyl-N,N',N''-triacetylchitotriose | Saccharomyces cerevisiae |
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 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 |