Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3 pLysS), subcloning in strain DH5alpha | Cellulomonas fimi |
Protein Variants | Comment | Organism |
---|---|---|
A211D | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
A253S | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
D287C | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
D287E | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
D287N | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
E368A | site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme | Cellulomonas fimi |
E368A/E407A | site-directed mutagenesis, the double mutant shows decreased activity at pH 5.0 compared to the wild-type enzyme | Cellulomonas fimi |
E407A | site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme | Cellulomonas fimi |
K292A | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
K292A/E407A | site-directed mutagenesis, the double mutant shows increased activity at pH 5.0 compared to the wild-type enzyme | Cellulomonas fimi |
L387P | site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme | Cellulomonas fimi |
additional information | engineering of cellulase A from Cellulomonas fimi as a model to replace residues that potentially influence the pH-activity profile of the enzyme based on sequence alignments and analysis of the known three-dimensional structures of other CAZy family 6 glycoside hydrolases with the aim to lower its pH optimum, overview | Cellulomonas fimi |
N320C | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
N320D | site-directed mutagenesis, the mutant shows highly reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
N320E | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
N360D | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
N360H | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
N360K | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
N360R | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
Q256C | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
Q256D | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
Q256E | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
S319A | site-directed mutagenesis, the mutant shows reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
S319D | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
S319H | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
S319K | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
S319R | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
Y321A | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
Y321F | site-directed mutagenesis, the mutant shows reduced activity at pH values of pH 7 and pH 9, but increased activity at pH 5, compared to the wild-type enzyme | Cellulomonas fimi |
Y321F/E407A | site-directed mutagenesis, the double mutant shows increased activity at pH 5.0 compared to the wild-type enzyme | Cellulomonas fimi |
Y321H | site-directed mutagenesis, the mutant shows highly reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
Y321K | site-directed mutagenesis, almost inactive mutant, residual activity ta pH 7.0 | Cellulomonas fimi |
Y321R | site-directed mutagenesis, the mutant shows highly reduced activity at different pH values compared to the wild-type enzyme | Cellulomonas fimi |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
carboxymethyl cellulose + H2O | Cellulomonas fimi | - |
? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cellulomonas fimi | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
carboxymethyl cellulose + H2O | - |
Cellulomonas fimi | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
cellulase A | - |
Cellulomonas fimi |
CenA | - |
Cellulomonas fimi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Cellulomonas fimi |