Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli ER2566 | Caldicellulosiruptor saccharolyticus |
Protein Variants | Comment | Organism |
---|---|---|
E359A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The E359A mutant exhibits no activity | Caldicellulosiruptor saccharolyticus |
F349A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The F349A, R111A, and W187A mutant enzyme in the presence of galactose shows remaining activity of 65% | Caldicellulosiruptor saccharolyticus |
F349S | mutant exhibits the highest activity in the presence of galactose relative to the activity measured in the absence of galactose among the tested mutant enzymes at position 349. The Ki-value for galactose of the F349S mutant (160 mM), which is 13fold that of the wild-type enzyme, is the highest among the reported values of beta-galactosidase. The wild-type enzyme hydrolyzes 62% of 100 g lactose/l with the addition of 30 g galactose/l, whereas the F349S mutant hydrolyzes more than 99% | Caldicellulosiruptor saccharolyticus |
H362A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The H362A mutant in the presence of galactose exhibits remaining activity below 30% | Caldicellulosiruptor saccharolyticus |
I44A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The I44A mutant in the presence of galactose exhibits remaining activity below 30% | Caldicellulosiruptor saccharolyticus |
N149A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The N149A mutant in the presence of galactose exhibits remaining activity below 30% | Caldicellulosiruptor saccharolyticus |
R111A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The F349A, R111A, and W187A mutant enzyme in the presence of galactose shows remaining activity of 46% | Caldicellulosiruptor saccharolyticus |
S41A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The S41A mutant in the presence of galactose exhibits remaining activity below 30% | Caldicellulosiruptor saccharolyticus |
W187A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The F349A, R111A, and W187A mutant enzyme in the presence of galactose shows remaining activity of 44% | Caldicellulosiruptor saccharolyticus |
W319A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The W319A mutant in the presence of galactose exhibits remaining activity below 30% | Caldicellulosiruptor saccharolyticus |
Y281A | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The Y281A mutant exhibits no activity | Caldicellulosiruptor saccharolyticus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
D-galactose | 30 g galactose/l. The wild-type enzyme exhibits 13% remaining activity relative to the activity observed in the absence of galactose. The Y281A and E359A mutants exhibit no activity and the S41A, I44A, N149A, W319A, and H362A mutants in the presence of galactose exhibit remaining activity below 30%. The F349A, R111A, and W187A mutant enzymes in the presence of galactose show remaining activity of 65, 46, and 44%, respectively | Caldicellulosiruptor saccharolyticus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
80000 | - |
- |
Caldicellulosiruptor saccharolyticus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Caldicellulosiruptor saccharolyticus | A4XI97 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Caldicellulosiruptor saccharolyticus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl beta-D-galactopyranoside + H2O | - |
Caldicellulosiruptor saccharolyticus | 4-nitrophenol + beta-D-galactopyranose | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
assay at | Caldicellulosiruptor saccharolyticus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
12 | - |
D-galactose | pH 6.0, 80°C, wild-type enzyme | Caldicellulosiruptor saccharolyticus | |
70 | - |
D-galactose | pH 6.0, 80°C, mutant enzyme F349V | Caldicellulosiruptor saccharolyticus | |
147 | - |
D-galactose | pH 6.0, 80°C, mutant enzyme F349L | Caldicellulosiruptor saccharolyticus | |
156 | - |
D-galactose | pH 6.0, 80°C, mutant enzyme F349A | Caldicellulosiruptor saccharolyticus | |
160 | - |
D-galactose | pH 6.0, 80°C, mutant enzyme F349S | Caldicellulosiruptor saccharolyticus |