Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Buedenbender, S.; Schulz, G.E.
    Structural base for enzymatic cyclodextrin hydrolysis (2009), J. Mol. Biol., 385, 606-617.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.54 expression in Escherichia coli BL21 (DE3) Flavobacterium sp. 92

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.54 mutant E340Q alpha-cyclodextrin complex, 0.1 M HEPES, 3.8 M NaCl, 3 mM CaCl2, pH 7.5, vapor diffusion, hanging drop, temperature 20°C, X-ray diffraction structure determination and analysis at 2.65 A resolution, space group H 3 2, T79P contact mutant, 0.1 M Tris, 17% PEG1500, pH 8.75, vapor diffusion, hanging drop, temperature 20°C, X-ray diffraction structure determination and analysis at 1.71 A resolution, space group P 21 21 2, mutant T49P, E340Q alpha-cyclodextrin complex, 0.1 M Tris, 17% PEG1500, pH 8.75, vapor diffusion, hanging drop, temperature 20°C, X-ray diffraction structure determination and analysis at 1.65 A resolution, space group P 21 21 2, mutant T49P, E340Q beta-cyclodextrin complex, resolution 1.69 A, mutant T49P, E340Q gamma-cyclodextrin complex, resolution 1.77 A Flavobacterium sp. 92

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.54 E340Q The active center mutant E340Q yields as much purified protein as the wild-type, and shows a residual, though very low, activity for alpha-cyclomaltodextrinase as derived from the crystal structure analysis Flavobacterium sp. 92
3.2.1.54 G523R molucular weight 182000, does not show a clear dimer mass Flavobacterium sp. 92
3.2.1.54 additional information the double mutant T49P-G523R does not show a clear monomer mass Flavobacterium sp. 92
3.2.1.54 R464A Arg464 is suggested to chaperone the substrates to the active center of the same enzyme subunit Flavobacterium sp. 92
3.2.1.54 R464S Arg464 is suggested to chaperone the substrates to the active center of the same enzyme subunit Flavobacterium sp. 92
3.2.1.54 T49P molucular weight 132000, mutant T49P, which causes the dissociation of the tetrameric wild-type FspCMD to the tight dimers, reduces the activity to less than 50% with a particularly strong reduction for beta-cyclomaltodextrinase and gamma-cyclomaltodextrinase Flavobacterium sp. 92

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.54 2.2
-
 gamma-cyclodextrin wild-type Flavobacterium sp. 92
3.2.1.54 2.9
-
 beta-cyclodextrin wild-type Flavobacterium sp. 92
3.2.1.54 4.1
-
 alpha-cyclodextrin wild-type Flavobacterium sp. 92

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.54 132000
-
 mutant enzyme T49P Flavobacterium sp. 92
3.2.1.54 138000
-
 double mutant T49P-G523R Flavobacterium sp. 92
3.2.1.54 182000
-
 mutant enzyme G523R Flavobacterium sp. 92
3.2.1.54 209000
-
 wild-type, size-exclusion chromatography Flavobacterium sp. 92

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.54 Flavobacterium sp. 92 Q8KKG0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.54 ion-exchange chromatography Flavobacterium sp. 92

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.54 36.1
-
 wild-type, alpha-cyclodextrin Flavobacterium sp. 92

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.54 alpha-cyclodextrin + H2O
-
 Flavobacterium sp. 92 ?
-
 ?
3.2.1.54 beta-cyclodextrin + H2O
-
 Flavobacterium sp. 92 ?
-
 ?
3.2.1.54 gamma-cyclodextrin + H2O
-
 Flavobacterium sp. 92 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.54 tetramer a loose dimer of tight dimers Flavobacterium sp. 92

Synonyms

EC Number Synonyms Comment Organism
3.2.1.54 CMD
-
 Flavobacterium sp. 92
3.2.1.54 cyclomaltodextrinase
-
 Flavobacterium sp. 92
3.2.1.54 FspCMD
-
 Flavobacterium sp. 92

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.54 17.4
-
 gamma-cyclodextrin wild-type Flavobacterium sp. 92
3.2.1.54 26.5
-
 beta-cyclodextrin wild-type Flavobacterium sp. 92
3.2.1.54 40.9
-
 alpha-cyclodextrin wild-type Flavobacterium sp. 92