Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Zobellia galactanivorans |
expressed in Escherichia coli C43(DE3) cells | Pseudoalteromonas carrageenovora |
Crystallization (Comment) | Organism |
---|---|
hanging drop vapor diffusion method, 28-29% PEG 3350, 100 mM MES, pH 6.5, and 0.3 M NaNO3 | Zobellia galactanivorans |
mutant enzyme E168D in complex with kappa-neocarratetraose, hanging drop vapor diffusion method, using 1.0 M sodium citrate and 100 mM cacodylate at pH 6.5 | Pseudoalteromonas carrageenovora |
Protein Variants | Comment | Organism |
---|---|---|
E168D | inactive | Pseudoalteromonas carrageenovora |
Q171A | the mutant shows 157% of wild type activity | Pseudoalteromonas carrageenovora |
R151A | the mutant shows 199% of wild type activity | Pseudoalteromonas carrageenovora |
R196A | the mutant shows 3% of wild type activity | Pseudoalteromonas carrageenovora |
R260A | the mutant shows less than 1% of wild type activity | Pseudoalteromonas carrageenovora |
R92A | the mutant shows 102% of wild type activity | Pseudoalteromonas carrageenovora |
W266A | the mutant shows 5% of wild type activity | Pseudoalteromonas carrageenovora |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000385 | - |
kappa-carrageen | mutant enzyme Q171A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
0.000683 | - |
kappa-carrageen | wild type enzyme, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
0.000822 | - |
kappa-carrageen | wild type enzyme, at pH 7.0 and 40°C | Zobellia galactanivorans | |
0.000899 | - |
kappa-carrageen | mutant enzyme R92A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
0.00115 | - |
kappa-carrageen | mutant enzyme R196A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
0.00121 | - |
kappa-carrageen | mutant enzyme R151A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
0.00263 | - |
kappa-carrageen | mutant enzyme W266A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
kappa-carrageen + H2O | Zobellia galactanivorans | - |
kappa-neocarrahexaose + kappa-neocarratetraose | - |
? | |
kappa-carrageen + H2O | Pseudoalteromonas carrageenovora | - |
kappa-neocarratetraose + kappa-neocarrabiose | - |
? | |
kappa-carrageen + H2O | Pseudoalteromonas carrageenovora ATCC 43555T | - |
kappa-neocarratetraose + kappa-neocarrabiose | - |
? | |
kappa-carrageen + H2O | Zobellia galactanivorans DsijT | - |
kappa-neocarrahexaose + kappa-neocarratetraose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudoalteromonas carrageenovora | P43478 | - |
- |
Pseudoalteromonas carrageenovora ATCC 43555T | P43478 | - |
- |
Zobellia galactanivorans | G0L921 | - |
- |
Zobellia galactanivorans DsijT | G0L921 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni affinity column chromatography and Sephadex gel filtration | Pseudoalteromonas carrageenovora |
Ni affinity column chromatography and Sephadex gel filtration | Zobellia galactanivorans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
kappa-carrageen + H2O | - |
Zobellia galactanivorans | kappa-neocarrahexaose + kappa-neocarratetraose | - |
? | |
kappa-carrageen + H2O | - |
Zobellia galactanivorans DsijT | kappa-neocarrahexaose + kappa-neocarratetraose | - |
? | |
kappa-carrageen + H2O | - |
Pseudoalteromonas carrageenovora | kappa-neocarratetraose + kappa-neocarrabiose | - |
? | |
kappa-carrageen + H2O | - |
Pseudoalteromonas carrageenovora ATCC 43555T | kappa-neocarratetraose + kappa-neocarrabiose | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CgkA | - |
Pseudoalteromonas carrageenovora |
CgkA | - |
Zobellia galactanivorans |
CgkAGH16 | - |
Pseudoalteromonas carrageenovora |
CgkAGH16 | - |
Zobellia galactanivorans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
6.13 | - |
kappa-carrageen | mutant enzyme R196A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
27.36 | - |
kappa-carrageen | mutant enzyme W266A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
120.17 | - |
kappa-carrageen | mutant enzyme Q171A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
135.84 | - |
kappa-carrageen | wild type enzyme, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
183.2 | - |
kappa-carrageen | mutant enzyme R92A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
478.79 | - |
kappa-carrageen | mutant enzyme R151A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
914.65 | - |
kappa-carrageen | wild type enzyme, at pH 7.0 and 40°C | Zobellia galactanivorans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5310 | - |
kappa-carrageen | mutant enzyme R196A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
10400 | - |
kappa-carrageen | mutant enzyme W266A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
199000 | - |
kappa-carrageen | wild type enzyme, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
204000 | - |
kappa-carrageen | mutant enzyme R92A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
312000 | - |
kappa-carrageen | mutant enzyme Q171A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
397000 | - |
kappa-carrageen | mutant enzyme R151A, at pH 7.0 and 40°C | Pseudoalteromonas carrageenovora | |
1110000 | - |
kappa-carrageen | wild type enzyme, at pH 7.0 and 40°C | Zobellia galactanivorans |