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Literature summary extracted from

  • Jensen, M.S.; Fredriksen, L.; MacKenzie, A.K.; Pope, P.B.; Leiros, I.; Chylenski, P.; Williamson, A.K.; Christopeit, T.; Ostby, H.; Vaaje-Kolstad, G.; Eijsink, V.G.H.
    Discovery and characterization of a thermostable two-domain GH6 endoglucanase from a compost metagenome (2018), PLoS ONE, 13, e0197862 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.4 gene mgCel6A, recombinant expression of His-tagged wild-type and mutant enzyme from codon-optimized gene in Escherichia coli strain TOP10 uncultured bacterium

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.4 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

EC Number Protein Variants Comment Organism
3.2.1.4 additional information construction of mutant mgCel6ADELTACBM lacking the carbohydrate binding module 2 (CBM2) uncultured bacterium

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.4 uncultured bacterium
-
 isolated from a compost metagenome
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.4 recominant His-tagged wild-type and mutant enzyme from Escherichia coli strain TOP10 by nickel affinity chromatography and ultrafiltration uncultured bacterium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.4 avicel + H2O substrate concentration 1%, low activity uncultured bacterium cellobiose + cellotriose
-
 ?
3.2.1.4 carboxymethyl cellulose + H2O substrate concentration 1%, low activity uncultured bacterium cellobiose + cellotriose
-
 ?
3.2.1.4 konjac glucomannan + H2O substrate concentration 1% uncultured bacterium cellobiose + cellotriose
-
 ?
3.2.1.4 additional information he later dominance of the dimeric product suggests that the trimer is slowly converted uncultured bacterium ?
-
 ?
3.2.1.4 phosphoric acid swollen cellulose + H2O substrate concentration 1%, best substrate and high activity uncultured bacterium cellobiose + cellotriose
-
 ?
3.2.1.4 sulfite-pulped spruce + H2O substrate concentration 1% uncultured bacterium cellobiose + cellotriose
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.4 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

EC Number Synonyms Comment Organism
3.2.1.4 GH6 endoglucanase
-
 uncultured bacterium
3.2.1.4 mgCel6A
-
 uncultured bacterium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.4 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 [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.4 76
-
 Tm of enzyme MgCel6A uncultured bacterium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.4 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

EC Number General Information Comment Organism
3.2.1.4 evolution the enzyme belongs to the glycoside hydrolase family 6, GH6 uncultured bacterium
3.2.1.4 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
3.2.1.4 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