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Literature summary extracted from

  • Brunecky, R.; Alahuhta, M.; Bomble, Y.J.; Xu, Q.; Baker, J.O.; Ding, S.Y.; Himmel, M.E.; Lunin, V.V.
    Structure and function of the Clostridium thermocellum cellobiohydrolase A X1-module repeat: enhancement through stabilization of the CbhA complex (2012), Acta Crystallogr. Sect. D, 68, 292-299.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.91 expressed in Escherichia coli BL21 cells Acetivibrio thermocellus
3.2.1.176 recombinant expression of His6-tagged enzyme in Escherichia coli (BL21) Acetivibrio thermocellus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.91 sitting drop vapor diffusion method, using in 0.2 M ammonium iodide, 20% (w/v) polyethylene glycol 3350 pH 6.2 or 0.2 M CaCl2, 20% (w/v) polyethylene glycol 3350 pH 5.1 Acetivibrio thermocellus
3.2.1.176 purified recombinant enzyme, sitting drop vapor diffusion method, mixing of 0.001 ml each of 80 mg/ml protein, in 20 mM Tris, pH 7.0, with 100 mM NaCl, and 1.9 M sodium malonate, pH 6.0, or with 1.9 M sodium malonate, pH 6.5, 20% v/v ethylene glycol, 50 mM KI. In a second method, 15 mg/ml protein in 20 mM acetic acid buffer, pH 5.0, with 100 mM NaCl is mixed with 0.2 M ammonium iodide, 20% w/v PEG 3350, pH 6.2, X-ray diffraction structure determination and analysis at 1.7 A resolution, single isomorphous replacement with anomalous signal from iodine and molecular-replacement Acetivibrio thermocellus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.91 Ca2+ contains Ca2+ Acetivibrio thermocellus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.91 cellulose + H2O Acetivibrio thermocellus
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.91 Acetivibrio thermocellus
-
-
-
3.2.1.176 Acetivibrio thermocellus A3DCH2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.91 Ni-NTA column chromatography, Source 15S column chromatography, and Superdex 75 gel filtration Acetivibrio thermocellus
3.2.1.176 recombinant His6-tagged enzyme from Escherichia coli (BL21) by cation exchange and hydrophobic interaction chromatography, followed by another step of cation exchange and anion exchange chromatography and gel filtration Acetivibrio thermocellus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.91 cellulose + H2O
-
 Acetivibrio thermocellus ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.91 CbhA
-
 Acetivibrio thermocellus
3.2.1.91 cellobiohydrolase A
-
 Acetivibrio thermocellus
3.2.1.176 CbhA
-
 Acetivibrio thermocellus
3.2.1.176 cellobiohydrolase A
-
 Acetivibrio thermocellus

General Information

EC Number General Information Comment Organism
3.2.1.176 metabolism cellulosomal enzyme systems utilize self-assembled scaffolded multimodule enzymes to deconstruct biomass, structure-function relationships governing the action of the large cellulosomal enzyme complex Acetivibrio thermocellus
3.2.1.176 additional information crystal structures of the two X1 modules from the enzyme CbhA Acetivibrio thermocellus