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Literature summary for 2.4.1.19 extracted from

  • Goh, P.H.; Illias, R.M.; Goh, K.M.
    Rational mutagenesis of cyclodextrin glucanotransferase at the calcium binding regions for enhancement of thermostability (2012), Int. J. Mol. Sci., 13, 5307-5323.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus sp. (in: Bacteria)

Protein Variants

Protein Variants Comment Organism
N132R introduction of an ionic interaction at the first Ca2+ site, disruption of catalytic activity Bacillus sp. (in: Bacteria)
N28R introduction of an additional ionic interaction at the second Ca2+ site, mutant displays increased cyclization activity Bacillus sp. (in: Bacteria)
S182E mutation adjacent to the first Ca2+ site and the active site cleft. Mutant shows enhanced thermostability, and decreased catalytic activity Bacillus sp. (in: Bacteria)
S182G mutation adjacent to the first Ca2+ site and the active site cleft. Increase in half-life at 60°C to 94 min Bacillus sp. (in: Bacteria)

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
75000
-
x * 75000, SDS-PAGE Bacillus sp. (in: Bacteria)

Organism

Organism UniProt Comment Textmining
Bacillus sp. (in: Bacteria) Q5U9V9
-
-
Bacillus sp. (in: Bacteria) G1 Q5U9V9
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.08
-
synthesis of alpha-cyclodextrin, 60°C, pH 6.0 Bacillus sp. (in: Bacteria)
216
-
synthesis of beta-cyclodextrin, 60°C, pH 6.0 Bacillus sp. (in: Bacteria)

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
soluble starch
-
Bacillus sp. (in: Bacteria) beta-cyclodextrin main product ?
soluble starch
-
Bacillus sp. (in: Bacteria) G1 beta-cyclodextrin main product ?

Subunits

Subunits Comment Organism
? x * 75000, SDS-PAGE Bacillus sp. (in: Bacteria)

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60
-
half-life 22 min for wild-type, 94 min for mutant S182G, 31 min for mutant S182E Bacillus sp. (in: Bacteria)

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.6
-
soluble starch mutant NS182G, pH 6.0, 60°C, calculated as production of beta-cyclodextrin Bacillus sp. (in: Bacteria)
2.6
-
soluble starch mutant S182E, pH 6.0, 60°C, calculated as production of beta-cyclodextrin Bacillus sp. (in: Bacteria)
3.13
-
soluble starch wild-type, pH 6.0, 60°C, calculated as production of beta-cyclodextrin Bacillus sp. (in: Bacteria)
3.5
-
soluble starch mutant N28R, pH 6.0, 60°C, calculated as production of beta-cyclodextrin Bacillus sp. (in: Bacteria)

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
additional information
-
additional information kcat/Km value for wild-type is 1.82 mg/ml, pH 6.0, 60°C Bacillus sp. (in: Bacteria)