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Literature summary for 3.2.1.91 extracted from

  • Kataeva, I.A.; Uversky, V.N.; Brewer, J.M.; Schubot, F.; Rose, J.P.; Wang, B.C.; Ljungdahl, L.G.
    Interactions between immunoglobulin-like and catalytic modules in Clostridium thermocellum cellulosomal cellobiohydrolase CbhA (2004), Protein Eng. Des. Sel., 17, 759-769.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene cbhA, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Acetivibrio thermocellus

Protein Variants

Protein Variants Comment Organism
D262A site-directed mutagenesis, mutation of the the module interface residue affects the interaction of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme Acetivibrio thermocellus
D264A site-directed mutagenesis, mutation of the the module interface residue affects the interaction of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme Acetivibrio thermocellus
additional information elimination of the enzyme's immunoglobulin-like module leads to its complete inactivation Acetivibrio thermocellus
T230A site-directed mutagenesis, mutation of the module interface residue affects the final fold and stability of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme Acetivibrio thermocellus
T230A/D262A site-directed mutagenesis, mutation of the residues affects the interaction of immunoglobulin-like module and the catalytic module, the mutant shows similar activity, but reduced stability and an altered mechanism in thermal unfolding compared to the wild-type enzyme Acetivibrio thermocellus

Localization

Localization Comment Organism GeneOntology No. Textmining
cellulosome
-
Acetivibrio thermocellus 43263
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ 2.45 mol bound by one pair of immunoglobulin-like module and catalytic module in the wild-type enzyme, the metal content is similar in the mutant enzymes, overview Acetivibrio thermocellus

Organism

Organism UniProt Comment Textmining
Acetivibrio thermocellus Q59325 gene cbhA
-
Acetivibrio thermocellus JW20 Q59325 gene cbhA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) to near homogeneity by nickel affinity chromatography and gel filtration Acetivibrio thermocellus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl cellobioside + H2O
-
Acetivibrio thermocellus 4-nitrophenol + cellobiose
-
?
4-nitrophenyl cellobioside + H2O
-
Acetivibrio thermocellus JW20 4-nitrophenol + cellobiose
-
?

Subunits

Subunits Comment Organism
More the enzyme is composed of an N-terminal carbohydrate-binding module of family 4, an immunoglobulin-like module, a catalytic module of glycoside hydrolase family 9, X1 1 and X1 2 modules, a carbohydrate-binding module of family 3 and finally a C-terminal dockerin module, elimination of the immunoglobulin-like module leads to complete inactivation of the catalytic module, intramolecular interaction analysis, residues T230, D262, and D264 are important, analysis of secondary and tertiary structure of the enzyme, overview Acetivibrio thermocellus

Synonyms

Synonyms Comment Organism
CbhA
-
Acetivibrio thermocellus
cellobiohydrolase
-
Acetivibrio thermocellus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
assay at Acetivibrio thermocellus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
additional information
-
thermodynamics and kinetics of the immunoglobulin-like module/catalytic module pair, wild-type and mutant enzymes, overview Acetivibrio thermocellus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
thermal unfolding kinetics of wild-type and mutant enzymes, overview Acetivibrio thermocellus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
assay at Acetivibrio thermocellus