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Literature summary extracted from

  • Santos, F.C.D.; Barbosa-Tessmann, I.P.
    Recombinant expression, purification, and characterization of a cyclodextrinase from Massilia timonae (2019), Protein Expr. Purif., 154, 74-84 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.54 expressed in Escherichia coli BL21 Star(DE3) cells Massilia timonae

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.54 CTAB complete inhibition at 5 mM Massilia timonae
3.2.1.54 Cu2+ 3.6% residual activity at 5 mM Massilia timonae
3.2.1.54 EDTA complete inhibition at 5 mM Massilia timonae
3.2.1.54 Fe3+ 13.9% residual activity at 5 mM Massilia timonae
3.2.1.54 K+ 78% residual activity at 10 mM Massilia timonae
3.2.1.54 Mg2+ 60.7% residual activity at 5 mM, complete inhibition at 10 mM Massilia timonae
3.2.1.54 Mn2+ 76.2% residual activity at 5 mM, complete inhibition at 10 mM Massilia timonae
3.2.1.54 Na+ 65.9% residual activity at 10 mM Massilia timonae
3.2.1.54 SDS complete inhibition at 5 mM Massilia timonae
3.2.1.54 Tween 80 62.1% residual activity at 1.0% (v/v) Massilia timonae
3.2.1.54 Zn2+ complete inhibition at 5 mM Massilia timonae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.54 2.1
-
 beta-cyclodextrin at pH 7.5 and 40°C Massilia timonae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.54 Ca2+ 276.8% activity at 5 mM Massilia timonae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.54 260000
-
 gel filtration Massilia timonae

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.54 Massilia timonae K9DE84
-
-
3.2.1.54 Massilia timonae CCUG 45783 K9DE84
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.54 HisTrap nickel column chromatography Massilia timonae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.54 beta-cyclodextrin + H2O 100% activity Massilia timonae D-glucose + maltose
-
 ?
3.2.1.54 maltodextrin + H2O 62% activity compared to beta-cyclodextrin Massilia timonae ?
-
 ?
3.2.1.54 maltodextrin + H2O 62% activity compared to beta-cyclodextrin Massilia timonae CCUG 45783 ?
-
 ?
3.2.1.54 maltoheptaose + H2O
-
 Massilia timonae D-glucose + maltose + ?
-
 ?
3.2.1.54 maltoheptaose + H2O
-
 Massilia timonae CCUG 45783 D-glucose + maltose + ?
-
 ?
3.2.1.54 maltotetraose + H2O
-
 Massilia timonae 2 D-glucose + maltose
-
 ?
3.2.1.54 maltotetraose + H2O
-
 Massilia timonae CCUG 45783 2 D-glucose + maltose
-
 ?
3.2.1.54 additional information no activity with maltose Massilia timonae ?
-
 ?
3.2.1.54 additional information no activity with maltose Massilia timonae CCUG 45783 ?
-
 ?
3.2.1.54 starch + H2O 8% activity compared to beta-cyclodextrin Massilia timonae ?
-
 ?
3.2.1.54 starch + H2O 8% activity compared to beta-cyclodextrin Massilia timonae CCUG 45783 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.54 homotetramer 4 * 65000, SDS-PAGE Massilia timonae
3.2.1.54 homotetramer 4 * 69400, calculated from amino acid sequence Massilia timonae

Synonyms

EC Number Synonyms Comment Organism
3.2.1.54 cyclodextrinase
-
 Massilia timonae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.54 40
-
-
 Massilia timonae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.54 25 50 the enzyme remains stable after 30 min incubation at 45°C and loses all its activity when incubated for 30 min at 50°C. The temperature at which the enzyme loses 50% of its activity with 2 mM calcium is 48.5°C. The enzyme remains 60% active after 24 h at 25 or 40°C but is 20% active at 0°C Massilia timonae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.54 138.8
-
 beta-cyclodextrin at pH 7.5 and 40°C Massilia timonae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.54 7
-
-
 Massilia timonae

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.54 6.5 8 the enzyme remains 100% active when incubated at pH values 6.5 and 8.0 after 24 h of incubation, and is 60% active when incubated at pH 7.0 for 24 h Massilia timonae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.54 68.3
-
 beta-cyclodextrin at pH 7.5 and 40°C Massilia timonae