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Literature summary extracted from
- Construction of a chimeric thermoacidophilic beta-endoglucanase (2013), BMC Biochem., 14, 11.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 3.2.1.4 | expression in Escherichia coli | Saccharolobus solfataricus |
| 3.2.1.4 | expression of chimeric enzymes in Escherichia coli. The proteins are produced in high yields in but aggregated in inclusion bodies.The fusion protein SSO1949-CelA-SSO1949, that consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949, is inactive with the substrate N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate. The fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, shows activity with the substrate N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate that is comparable to the native enzyme from Sulfolobus and shows a temperature optimum of approximately 85°C and a pH optimum of approximately pH 3 thus retaining the advantageous properties of the Sulfolobus parent enzyme | Thermotoga maritima |
| 3.2.1.4 | expression of chimeric enzymes in Escherichia coli. The proteins are produced in high yields in but aggregated in inclusion bodies.The fusion protein SSO1949-CelA-SSO1949, that consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949, is inactive with the substrate N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulphonate. The fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, shows activity with the substrate N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulphonate that is comparable to the native enzyme from Sulfolobus and shows a temperature optimum of approximately 85°C and a pH optimum of approximately pH 3 thus retaining the advantageous properties of the Sulfolobus parent enzyme | Saccharolobus solfataricus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 3.2.1.4 | additional information | after bioinformatic analysis hybrid proteins based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima are constructed which should successfully combine the advantageous properties of both cellulases, i.e. recombinant expression in Escherichia coli, acidophily and thermophily. both hybrids are expressed insoluble in Escherichia coli, but one hybrid enzyme was successfully refolded from washed inclusion bodies | Saccharolobus solfataricus |
| 3.2.1.4 | additional information | targeted construction of chimeric enzymes of cellulase from Sulfolobus solfataricus P2 and cellulase CelA from Thermotoga maritima. The fusion protein SSO1949-CelA-SSO1949 consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949. The catalytic center with the two catalytic glutamate residues is derived from SSO1949 whereas the reducing end of the substrate binding cleft comes from CelA. The fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA. Most of the active center is derived from SSO1949 | Thermotoga maritima |
| 3.2.1.4 | additional information | targeted construction of chimeric enzymes of cellulase from Sulfolobus solfataricus P2 and cellulase CelA from Thermotoga maritima. The fusion protein SSO1949-CelA-SSO1949 consists of 193 amino acids SSO1949 followed by 76 amino acids CelA and 39 amino acids SSO1949. The catalytic center with the two catalytic glutamate residues is derived from SSO1949 whereas the reducing end of the substrate binding cleft comes from CelA. The fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA. Most of the active center is derived from SSO1949 | Saccharolobus solfataricus |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.4 | 0.0017 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima |  |
| 3.2.1.4 | 0.0017 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus | |
| 3.2.1.4 | 0.00174 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH and temperature not specified in the publication, hybrid protein (CelA-SSO1949-CelA, consisting of 262 amino acids) based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima | Saccharolobus solfataricus | |
| 3.2.1.4 | 0.0055 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, native enzyme | Thermotoga maritima | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.4 | 29000 | - |
x * 29000, chimeric fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, SDS-PAGE | Thermotoga maritima |
| 3.2.1.4 | 29000 | - |
x * 29000, chimeric fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, SDS-PAGE | Saccharolobus solfataricus |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.2.1.4 | Saccharolobus solfataricus | Q97X08 | - |
- |
| 3.2.1.4 | Saccharolobus solfataricus P2 | Q97X08 | - |
- |
| 3.2.1.4 | Thermotoga maritima | Q60032 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.2.1.4 | - |
Saccharolobus solfataricus |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.4 | 0.8 | - |
substrate N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate, pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima |
| 3.2.1.4 | 0.8 | - |
substrate: N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate, pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus |
| 3.2.1.4 | 0.8 | - |
substrate: N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate, pH and temperature not specified in the publication, hybrid protein (CelA-SSO1949-CelA, consisting of 262 amino acids) based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima | Saccharolobus solfataricus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.2.1.4 | N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate + H2O | - |
Thermotoga maritima | ? | - |
? | |
| 3.2.1.4 | N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate + H2O | - |
Saccharolobus solfataricus | ? | - |
? | |
| 3.2.1.4 | N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate + H2O | - |
Saccharolobus solfataricus P2 | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.2.1.4 | ? | x * 29000, chimeric fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, SDS-PAGE | Thermotoga maritima |
| 3.2.1.4 | ? | x * 29000, chimeric fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, SDS-PAGE | Saccharolobus solfataricus |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 3.2.1.4 | SSO1949 | locus name | Saccharolobus solfataricus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.4 | 85 | - |
chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima |
| 3.2.1.4 | 85 | - |
chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus |
| 3.2.1.4 | 85 | - |
hybrid protein (CelA-SSO1949-CelA) based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima | Saccharolobus solfataricus |
| 3.2.1.4 | 95 | - |
native enzyme | Thermotoga maritima |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.4 | 75 | 99 | 75°C: about 50% of maximal activity, 99°C: about 60% of maximal activity. Chimeric fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, SDS-PAGE | Thermotoga maritima |
| 3.2.1.4 | 75 | 99 | 75°C: about 50% of maximal activity, 99°C: about 60% of maximal activity. Chimeric fusion protein CelA-SSO1949-CelA consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA, SDS-PAGE | Saccharolobus solfataricus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.4 | 0.39 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima | |
| 3.2.1.4 | 0.39 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus | |
| 3.2.1.4 | 0.39 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH and temperature not specified in the publication, hybrid protein (CelA-SSO1949-CelA, consisting of 262 amino acids) based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima | Saccharolobus solfataricus | |
| 3.2.1.4 | 19 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, native enzyme | Thermotoga maritima | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.4 | 3 | - |
chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima |
| 3.2.1.4 | 3 | - |
chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus |
| 3.2.1.4 | 3 | - |
hybrid protein (CelA-SSO1949-CelA) based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima | Saccharolobus solfataricus |
| 3.2.1.4 | 4.5 | - |
native enzyme | Thermotoga maritima |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.4 | 3.5 | 4.5 | pH 3.5: about 70% of maximal activity, pH 4.5: about 45% of maximal activity, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima |
| 3.2.1.4 | 3.5 | 4.5 | pH 3.5: about 70% of maximal activity, pH 4.5: about 45% of maximal activity, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.4 | 229 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Thermotoga maritima | |
| 3.2.1.4 | 229 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, chimeric fusion protein CelA-SSO1949-CelA, that consists of 70 amino acids CelA followed by 163 amino acids SSO1949 and 29 amino acids CelA | Saccharolobus solfataricus | |
| 3.2.1.4 | 230 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH and temperature not specified in the publication, hybrid protein (CelA-SSO1949-CelA, consisting of 262 amino acids) based on beta-endoglucanase from Sulfolobus solfataricus P2 and beta-endoglucanase from Thermotoga maritima | Saccharolobus solfataricus | |
| 3.2.1.4 | 3455 | - |
N-[2-N-[(S-(4-deoxy-4-dimethylamino-phenylazophenylthioureido-beta-D-glucopyranosyl)-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl-(1->4)-beta-D-glucopyranosyl)-2-thioacetyl]aminoethyl]-1-naphthylamine-5-sulfonate | pH 3, 80°C, native enzyme | Thermotoga maritima | |
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