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Literature summary extracted from

  • Cockburn, D.W.; Vandenende, C.; Clarke, A.J.
    Modulating the pH-activity profile of cellulase by substitution: replacing the general base catalyst aspartate with cysteinesulfinate in cellulase A from Cellulomonas fimi (2010), Biochemistry, 49, 2042-2050.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.4 CenA, DNA and amino acid sequence determination and analysis, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Cellulomonas fimi

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.4 D216C replacement of Asp with cysteinesulfinate by combination of site-directed mutagenesis and chemical modification, the substituted cysteinyl residue is oxidized to cysteine sulfinic acid with hydrogen peroxide, the resulting protein product retains its native structure, almost inactive mutant Cellulomonas fimi
3.2.1.4 D216N site-directed mutagenesis, the mutant shows reduced activity and a shift in pH dependence compared to the wild-type enzyme Cellulomonas fimi
3.2.1.4 D252C site-directed mutagenesis, substitution of the catalytic acid residue Asp252, almost inactive mutant Cellulomonas fimi
3.2.1.4 D392A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Cellulomonas fimi
3.2.1.4 D392C replacement of Asp with cysteinesulfinate by combination of site-directed mutagenesis and chemical modification, the substituted cysteinyl residue is oxidized to cysteine sulfinic acid with hydrogen peroxide, the resulting protein product retains its native structure. Oxidation of the Asp392Cys mutant enzyme restores 52% of wild-type activity when assessed at pH 7.5. The replacement of Asp392 with cysteine sulfinate induced an acidic shift in the pH profile of the enzyme such that this enzyme derivative is more active than wild-type CenA below pH 5.5 Cellulomonas fimi
3.2.1.4 D392N site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Cellulomonas fimi
3.2.1.4 D392S site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Cellulomonas fimi

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.2.1.4 extracellular CenA Cellulomonas fimi
-
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.4 46700
-
 x * 46700, CenA Cellulomonas fimi

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.4 cellulose + H2O Cellulomonas fimi
-
 ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.4 Cellulomonas fimi
-
 gene cenA
-

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.4 cellohexaose + H2O = 2 cellotriose proposed mechanism of action of inverting glycoside hydrolases and structures of catalytic residues, overview. The catalytic base is Asp392, catalytic acid residue is Asp252 Cellulomonas fimi
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.4 carboxymethyl cellulose + H2O
-
 Cellulomonas fimi ?
-
 ?
3.2.1.4 cellulose + H2O
-
 Cellulomonas fimi ?
-
 ?
3.2.1.4 additional information inverting glycoside hydrolases catalyze bond cleavage using a single-displacement mechanism involving the participation of two acidic amino acid residues positioned opposite each other across the active site cleft or tunnel Cellulomonas fimi ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.4 ? x * 46700, CenA Cellulomonas fimi

Synonyms

EC Number Synonyms Comment Organism
3.2.1.4 cellulase A
-
 Cellulomonas fimi
3.2.1.4 CenA
-
 Cellulomonas fimi
3.2.1.4 More cellulase A is an inverting glycoside hydrolase and a member of family 6 of the CAZy database classification system Cellulomonas fimi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.4 37
-
 assay at Cellulomonas fimi

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.4 0.0013
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D392A Cellulomonas fimi
3.2.1.4 0.0051
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D392S Cellulomonas fimi
3.2.1.4 0.0059
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D392N Cellulomonas fimi
3.2.1.4 0.0073
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D252C Cellulomonas fimi
3.2.1.4 0.014
-
 carboxymethyl cellulose pH 7.0, 37°C, oxidized mutant D252C Cellulomonas fimi
3.2.1.4 0.0146
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D216C Cellulomonas fimi
3.2.1.4 0.029
-
 carboxymethyl cellulose pH 7.0, 37°C, oxidized mutant D216C Cellulomonas fimi
3.2.1.4 0.227
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D216N Cellulomonas fimi
3.2.1.4 0.994
-
 carboxymethyl cellulose pH 7.0, 37°C, mutant D392C Cellulomonas fimi
3.2.1.4 3.57
-
 carboxymethyl cellulose pH 7.0, 37°C, oxidized mutant D392C Cellulomonas fimi
3.2.1.4 5.81
-
 carboxymethyl cellulose pH 7.0, 37°C, H2O2-treated wild-type CenA Cellulomonas fimi
3.2.1.4 5.92
-
 carboxymethyl cellulose pH 7.0, 37°C, wild-type CenA Cellulomonas fimi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.4 7 7.5 wild-type CenA Cellulomonas fimi

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.4 4 9 profiles of wild-type and mutant D392C and D216N enzymes, overview Cellulomonas fimi

General Information

EC Number General Information Comment Organism
3.2.1.4 metabolism the complete solubilization of crystalline cellulose requires the concerted and synergistic action of three classes of glycoside hydrolases, cellulase, endoglucanase, EC 3.2.1.4, cellobiohydrolase, EC 3.2.1.74, and beta-glucosidase, EC 3.2.1.21 Cellulomonas fimi