Cloned (Comment) | Organism |
---|---|
gene cbhA, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Acetivibrio thermocellus |
Protein Variants | Comment | Organism |
---|---|---|
D262A | site-directed mutagenesis, mutation of the the module interface residue affects the interaction of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme | Acetivibrio thermocellus |
D264A | site-directed mutagenesis, mutation of the the module interface residue affects the interaction of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme | Acetivibrio thermocellus |
additional information | elimination of the enzyme's immunoglobulin-like module leads to its complete inactivation | Acetivibrio thermocellus |
T230A | site-directed mutagenesis, mutation of the module interface residue affects the final fold and stability of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme | Acetivibrio thermocellus |
T230A/D262A | site-directed mutagenesis, mutation of the residues affects the interaction of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme | Acetivibrio thermocellus |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cellulosome | - |
Acetivibrio thermocellus | 43263 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 2.45 mol bound by one pair of immunoglobulin-like module and catalytic module in the wild-type enzyme, the metal content is similar in the mutant enzymes, overview | Acetivibrio thermocellus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Acetivibrio thermocellus | Q59325 | gene cbhA | - |
Acetivibrio thermocellus JW20 | Q59325 | gene cbhA | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) to near homogeneity by nickel affinity chromatography and gel filtration | Acetivibrio thermocellus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl cellobioside + H2O | - |
Acetivibrio thermocellus | 4-nitrophenol + cellobiose | - |
? | |
4-nitrophenyl cellobioside + H2O | - |
Acetivibrio thermocellus JW20 | 4-nitrophenol + cellobiose | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme is composed of an N-terminal carbohydrate-binding module of family 4, an immunoglobulin-like module, a catalytic module of glycoside hydrolase family 9, X1 1 and X1 2 modules, a carbohydrate-binding module of family 3 and finally a C-terminal dockerin module, elimination of the immunoglobulin-like module leads to complete inactivation of the catalytic module, intramolecular interaction analysis, residues T230, D262, and D264 are important, analysis of secondary and tertiary structure of the enzyme, overview | Acetivibrio thermocellus |
Synonyms | Comment | Organism |
---|---|---|
CbhA | - |
Acetivibrio thermocellus |
cellobiohydrolase | - |
Acetivibrio thermocellus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
65 | - |
assay at | Acetivibrio thermocellus |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermodynamics and kinetics of the immunoglobulin-like module/catalytic module pair, wild-type and mutant enzymes, overview | Acetivibrio thermocellus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
thermal unfolding kinetics of wild-type and mutant enzymes, overview | Acetivibrio thermocellus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
assay at | Acetivibrio thermocellus |