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Literature summary extracted from

  • Wang, H.J.; Hsiao, Y.Y.; Chen, Y.P.; Ma, T.Y.; Tseng, C.P.
    Polarity alteration of a calcium site induces a hydrophobic interaction network and enhances Cel9A endoglucanase thermostability (2016), Appl. Environ. Microbiol., 82, 1662-1674.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.203
-
 Alicyclobacillus acidocaldarius

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.203 crystal structure reveals that mutation D468A triggers a conformational change similar to that induced by Ca2+ in the wild type and develops a hydrophobic interaction network between the calcium site and the neighboring hydrophobic region (Ala113 to Ala117) Alicyclobacillus acidocaldarius

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.203 D468A calcium binding-site mutant, shows increased activity compared with the wild type with 0 mM or 10 mM Ca2+ added. The mutation triggers a conformational change similar to that induced by Ca2+ in the wild type. Mutant displays an increase in melting temperature by 5.9°C Alicyclobacillus acidocaldarius
3.2.1.203 D468E calcium binding-site mutant, kinetic parameters similar to wild-type Alicyclobacillus acidocaldarius
3.2.1.203 D468V calcium binding-site mutant, shows increased activity compared with the wild type with 0 mM or 10 mM Ca2+ added. Mutant displays an increase in melting temperature by 4.5°C Alicyclobacillus acidocaldarius
3.2.1.203 D469A calcium binding-site mutant, results in decreased activity Alicyclobacillus acidocaldarius
3.2.1.203 D469V calcium binding-site mutant, increase in kcat value Alicyclobacillus acidocaldarius
3.2.1.203 D471A calcium binding-site mutant, results in decreased activity Alicyclobacillus acidocaldarius
3.2.1.203 D471E calcium binding-site mutant, kinetic parameters similar to wild-type Alicyclobacillus acidocaldarius
3.2.1.203 D471V calcium binding-site mutant, increase in km value Alicyclobacillus acidocaldarius

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.203 additional information
-
 carboxymethyl cellulose Km value of wild-type, 29.95 mg/ml, mutant D468A, 46.8 mg/ml, mutant D469V 204.9 mg/ml, pH 5.5, 60°C Alicyclobacillus acidocaldarius

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.203 Ca2+ sequence contains a calcium-binding site (Ser465 to Val470) near the catalytic cleft. Ca2+ induces hydrophobic interactions between the calcium site and its nearby region by driving a conformational change. Calcium-binding site mutants D468A and D468V show increased activity compared with the wild type with 0 mM or 10 mM Ca2+ added Alicyclobacillus acidocaldarius

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.203 Alicyclobacillus acidocaldarius Q9AJS0
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.203 57
-
 mutant D469V, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 77
-
 mutant D469V, presence of 10 mM Ca2+, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 137
-
 wild-type, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 153
-
 mutant D468V, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 190
-
 mutant D468A, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 192
-
 wild-type, presence of 10 mM Ca2+, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 234
-
 mutant D468V, presence of 10 mM Ca2+, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 282
-
 mutant D468A, presence of 10 mM Ca2+, pH 5.5, 60°C Alicyclobacillus acidocaldarius

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.203 carboxymethyl cellulose + H2O
-
 Alicyclobacillus acidocaldarius ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.203 Cel9A
-
 Alicyclobacillus acidocaldarius
3.2.1.203 CelA
-
 Alicyclobacillus acidocaldarius

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.203 70
-
-
 Alicyclobacillus acidocaldarius

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.203 75.9
-
 wild-type, melting temperature Alicyclobacillus acidocaldarius
3.2.1.203 80.4
-
 mutant D468V, melting temperature Alicyclobacillus acidocaldarius
3.2.1.203 81.8
-
 mutant D468A, melting temperature Alicyclobacillus acidocaldarius

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.203 0.6
-
 carboxymethyl cellulose wild-type, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 0.68
-
 carboxymethyl cellulose mutant D468E, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 0.68
-
 carboxymethyl cellulose mutant D471V, pH 5.5, 60°C Alicyclobacillus acidocaldarius
3.2.1.203 0.72
-
 carboxymethyl cellulose mutant D471A, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 0.92
-
 carboxymethyl cellulose mutant D471E, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 1.31
-
 carboxymethyl cellulose mutant D468V, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 1.43
-
 carboxymethyl cellulose mutant D468A, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 1.43
-
 carboxymethyl cellulose mutant D469A, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 3.38
-
 carboxymethyl cellulose mutant D469V, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius
3.2.1.203 5.01
-
 carboxymethyl cellulose mutant D469E, pH 5.5, 60°C, calculated as amounts of reducing equivalents Alicyclobacillus acidocaldarius

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.203 5 5.5
-
 Alicyclobacillus acidocaldarius
3.2.1.203 6
-
 mutant D468A Alicyclobacillus acidocaldarius