Cloned (Comment) | Organism |
---|---|
gene mgCel6A, recombinant expression of His-tagged wild-type and mutant enzyme from codon-optimized gene in Escherichia coli strain TOP10 | uncultured bacterium |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged catalytic domain of the enzyme, sitting drop vapour diffusion method, mixing of 500 nl of 12 mg/ml protein solution with 500 nl of reservoir solution containing 1 M (NH4)2SO4, 0.1 M Bis-Tris, pH 5.5, and 1% PEG 3350, and equilibration against 0.06 ml of reservoir solution, 20°C, 5 weeks, X-ray diffraction structure determination and analysis at 1.88 A resolution, molecular replacement using the crystal structure of cellobiohydrolase Cel6A from Thermobifida fusca as search model (PDB ID 1TML) | uncultured bacterium |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of mutant mgCel6ADELTACBM lacking the carbohydrate binding module 2 (CBM2) | uncultured bacterium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
uncultured bacterium | - |
isolated from a compost metagenome | - |
Purification (Comment) | Organism |
---|---|
recominant His-tagged wild-type and mutant enzyme from Escherichia coli strain TOP10 by nickel affinity chromatography and ultrafiltration | uncultured bacterium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
avicel + H2O | substrate concentration 1%, low activity | uncultured bacterium | cellobiose + cellotriose | - |
? | |
carboxymethyl cellulose + H2O | substrate concentration 1%, low activity | uncultured bacterium | cellobiose + cellotriose | - |
? | |
konjac glucomannan + H2O | substrate concentration 1% | uncultured bacterium | cellobiose + cellotriose | - |
? | |
additional information | he later dominance of the dimeric product suggests that the trimer is slowly converted | uncultured bacterium | ? | - |
? | |
phosphoric acid swollen cellulose + H2O | substrate concentration 1%, best substrate and high activity | uncultured bacterium | cellobiose + cellotriose | - |
? | |
sulfite-pulped spruce + H2O | substrate concentration 1% | uncultured bacterium | cellobiose + cellotriose | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme consists of a glycoside hydrolase family 6 catalytic domain (GH6) and a family 2 carbohydrate binding module (CBM2) that are connected by a linker rich in prolines and threonines | uncultured bacterium |
Synonyms | Comment | Organism |
---|---|---|
GH6 endoglucanase | - |
uncultured bacterium |
mgCel6A | - |
uncultured bacterium |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
60 | - |
enzyme MgCel6A exhibits maximum activity at 85°C and pH 5.0 on carboxymethyl cellulose (CMC), but in prolonged incubations with the industrial substrate, the highest yields are obtained at 60°C, pH 6.0 | uncultured bacterium |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
76 | - |
Tm of enzyme MgCel6A | uncultured bacterium |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
enzyme MgCel6A exhibits maximum activity at 85°C and pH 5.0 on carboxymethyl cellulose (CMC), but in prolonged incubations with the industrial substrate, the highest yields are obtained at 60°C, pH 6.0 | uncultured bacterium |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycoside hydrolase family 6, GH6 | uncultured bacterium |
additional information | the enzyme consists of a glycoside hydrolase family 6 catalytic domain (GH6) and a family 2 carbohydrate binding module (CBM2) that are connected by a linker rich in prolines and threonines. Comparative studies with a truncated variant of the enzyme show that the CBM increases substrate binding, while not affecting thermal stability. Importantly, at higher substrate concentrations the full-length enzyme is outperformed by the catalytic domain alone, underpinning previous suggestions that CBMs may be less useful in high-consistency bioprocessing | uncultured bacterium |
physiological function | the GH6 endoglucanase mgCel6A is a thermostable cellulase with activity on an industrial cellulosic substrate (sulfite-pulped Norway spruce) and a glucomannanase side activity | uncultured bacterium |