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Literature summary extracted from

  • Ohshima, T.; Kawakami, R.; Kanai, Y.; Goda, S.; Sakuraba, H.
    Gene expression and characterization of 2-keto-3-deoxygluconate kinase, a key enzyme in the modified Entner-Doudoroff pathway of the aerobic and acidophilic hyperthermophile Sulfolobus tokodaii (2007), Protein Expr. Purif., 54, 73-78.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.45 expressed in Escherichia coli strain BL21(DE3) Sulfurisphaera tokodaii
2.7.1.45 expression in Escherichia coli Sulfurisphaera tokodaii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.45 0.027
-
 2-dehydro-3-deoxy-D-gluconate pH 8.5, 50°C Sulfurisphaera tokodaii
2.7.1.45 0.027
-
 2-dehydro-3-deoxy-D-gluconate in 50 mM Tris/HCl (pH 8.5) at 50°C Sulfurisphaera tokodaii
2.7.1.45 0.057
-
 ATP pH 8.5, 50°C Sulfurisphaera tokodaii
2.7.1.45 0.057
-
 ATP in 50 mM Tris/HCl (pH 8.5) at 50°C, with 2-dehydro-3-deoxy-D-gluconate as substrate Sulfurisphaera tokodaii
2.7.1.45 0.11
-
 ATP in 50 mM Tris/HCl (pH 8.5) at 50°C, with 2-keto-D-gluconate as substrate Sulfurisphaera tokodaii
2.7.1.45 1.3
-
 2-keto-D-gluconate pH 8.5, 50°C Sulfurisphaera tokodaii
2.7.1.45 1.3
-
 2-keto-D-gluconate in 50 mM Tris/HCl (pH 8.5) at 50°C Sulfurisphaera tokodaii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.45 Co2+ 10 mM, required for activity, 98% relative activity compared to Mg2+ Sulfurisphaera tokodaii
2.7.1.45 Co2+ 98% of the activity with Mg2+ Sulfurisphaera tokodaii
2.7.1.45 Mg2+ most effective Sulfurisphaera tokodaii
2.7.1.45 Mg2+ 10 mM, required for activity, most effective cation Sulfurisphaera tokodaii
2.7.1.45 Mn2+ 73% of the activity with Mg2+ Sulfurisphaera tokodaii
2.7.1.45 Mn2+ 10 mM, required for activity, 73% relative activity compared to Mg2+ Sulfurisphaera tokodaii
2.7.1.45 additional information Cu2+ and Ca2+ have no stimulatory effect on activity Sulfurisphaera tokodaii
2.7.1.45 additional information divalent kation is required for activity such as Mg2+, Co2+, Ni2+, Zn2+, Mn2+ Sulfurisphaera tokodaii
2.7.1.45 Ni2+ 92% of the activity with Mg2+ Sulfurisphaera tokodaii
2.7.1.45 Ni2+ 10 mM, required for activity, 92% relative activity compared to Mg2+ Sulfurisphaera tokodaii
2.7.1.45 Zn2+ 10 mM, required for activity, 75% relative activity compared to Mg2+ Sulfurisphaera tokodaii
2.7.1.45 Zn2+ 75% of the activity with Mg2+ Sulfurisphaera tokodaii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.45 30000
-
 4 * 30000, gel filtration Sulfurisphaera tokodaii
2.7.1.45 34480
-
 calculated from amino acid sequence Sulfurisphaera tokodaii
2.7.1.45 34480
-
 4 * 34480, calculated, 4 * 35000, SDS-PAGE Sulfurisphaera tokodaii
2.7.1.45 35000
-
 SDS-PAGE Sulfurisphaera tokodaii
2.7.1.45 35000
-
 4 * 34480, calculated, 4 * 35000, SDS-PAGE Sulfurisphaera tokodaii
2.7.1.45 120000
-
 gel filtration Sulfurisphaera tokodaii

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.45 Sulfurisphaera tokodaii
-
-
-
2.7.1.45 Sulfurisphaera tokodaii Q96XN9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.45 Butyl Toyopearl column chromatography and Superdex 200 column gel filtration Sulfurisphaera tokodaii
2.7.1.45 recombinant protein Sulfurisphaera tokodaii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.45 additional information
-
 development of spectrophotometric assay that uses three coupled reactions to measure activity. Assay displays a linear relationship between time and the increase in absorbance at 340 nm due to NADPH formation Sulfurisphaera tokodaii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.45 ATP + 2-dehydro-3-deoxy-D-gluconate
-
 Sulfurisphaera tokodaii ADP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?
2.7.1.45 ATP + 2-keto-D-gluconate
-
 Sulfurisphaera tokodaii ADP + 6-phospho-2-keto-D-gluconate
-
 ?
2.7.1.45 GTP + 2-dehydro-3-deoxy-D-gluconate 94% activity compared to the substrate combination with ATP Sulfurisphaera tokodaii GDP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?
2.7.1.45 GTP + 2-dehydro-3-deoxy-D-gluconate 94% of the activity with ATP Sulfurisphaera tokodaii GDP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?
2.7.1.45 ITP + 2-dehydro-3-deoxy-D-gluconate 66% activity compared to the substrate combination with ATP Sulfurisphaera tokodaii IDP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?
2.7.1.45 ITP + 2-dehydro-3-deoxy-D-gluconate 66% of the activity with ATP Sulfurisphaera tokodaii IDP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?
2.7.1.45 additional information does not accept ADP or AMP as phosphoryl donors Sulfurisphaera tokodaii ?
-
 ?
2.7.1.45 additional information no substrate: UTP, CTP, ADP, GDP, IDP, UDP, CDP, and AMP, D-glucose, D-fructose, and 2-keto-3-deoxyoctonate Sulfurisphaera tokodaii ?
-
 ?
2.7.1.45 TTP + 2-dehydro-3-deoxy-D-gluconate 43% activity compared to the substrate combination with ATP Sulfurisphaera tokodaii TDP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?
2.7.1.45 TTP + 2-dehydro-3-deoxy-D-gluconate 43% of the activity with ATP Sulfurisphaera tokodaii TDP + 6-phospho-2-dehydro-3-deoxy-D-gluconate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.45 homotetramer 4 * 30000, gel filtration Sulfurisphaera tokodaii
2.7.1.45 tetramer 4 * 34480, calculated, 4 * 35000, SDS-PAGE Sulfurisphaera tokodaii

Synonyms

EC Number Synonyms Comment Organism
2.7.1.45 2-keto-3-deoxygluconate kinase
-
 Sulfurisphaera tokodaii
2.7.1.45 KDGK
-
 Sulfurisphaera tokodaii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.45 80
-
-
 Sulfurisphaera tokodaii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.7.1.45 50
-
 65% of maximum activity Sulfurisphaera tokodaii
2.7.1.45 90
-
 25% of maximum activity Sulfurisphaera tokodaii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.7.1.45 50
-
 the activity at 50°C is about 65% of the maximum Sulfurisphaera tokodaii
2.7.1.45 80
-
 1 h, no loss of activity Sulfurisphaera tokodaii
2.7.1.45 80
-
 the purified recombinant enzyme is extremely thermostable and retains full activity after heating at 80°C for 1 h Sulfurisphaera tokodaii
2.7.1.45 85
-
 1 h, 50% residual activity Sulfurisphaera tokodaii
2.7.1.45 90
-
 the activity at 90°C is only about 25% of that at 80°C Sulfurisphaera tokodaii
2.7.1.45 90
-
 10 min, 10% residual activity Sulfurisphaera tokodaii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.45 8.26
-
 2-dehydro-3-deoxy-D-gluconate pH 8.5, 50°C Sulfurisphaera tokodaii
2.7.1.45 8.26
-
 2-dehydro-3-deoxy-D-gluconate in 50 mM Tris/HCl (pH 8.5) at 50°C Sulfurisphaera tokodaii
2.7.1.45 26.3
-
 2-keto-D-gluconate pH 8.5, 50°C Sulfurisphaera tokodaii
2.7.1.45 26.3
-
 2-keto-D-gluconate in 50 mM Tris/HCl (pH 8.5) at 50°C Sulfurisphaera tokodaii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.45 8.5
-
-
 Sulfurisphaera tokodaii

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.7.1.45 6.5 8.5 half of the maximum activity is obtained at pHs 6.5 and 9.5 Sulfurisphaera tokodaii

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.7.1.45 4.5
-
 no activity Sulfurisphaera tokodaii
2.7.1.45 10
-
 no activity Sulfurisphaera tokodaii

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.45 ATP
-
 Sulfurisphaera tokodaii