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Literature summary extracted from

  • Ye, X.; Rollin, J.; Zhang, Y.
    Thermophilic alpha-glucan phosphorylase from Clostridium thermocellum: cloning, characterization and enhanced thermostability (2010), J. Mol. Catal. B, 65, 110-116.
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.4.1.1 AMP slight activation Acetivibrio thermocellus
2.4.1.1 DTT activates Acetivibrio thermocellus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.1 ORF Cthe0357, DNA and amino acid sequence determination and analysis, expression of C-terminally His-tagged enzyme in Escherichia coli, expression of cellulose-binding module-intein-alphaGP fusion protein Acetivibrio thermocellus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.1 EDTA slight activation Acetivibrio thermocellus
2.4.1.1 Mn2+ strong inhibition at 1 mM Acetivibrio thermocellus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.1 0.054
-
 alpha-D-glucose 1-phosphate pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 0.3
-
 starch pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 0.39
-
 dextrin DP19 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 0.73
-
 dextrin DP14 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 1.2
-
 dextrin DP14 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 1.5
-
 starch pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 1.8
-
 dextrin DP4 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 1.9
-
 dextrin DP19 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 7.7
-
 phosphate pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 17.1
-
 dextrin DP4 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.1 Mg2+ activates at 1 mM Acetivibrio thermocellus
2.4.1.1 additional information the enzyme is not affected by 1 mM of Ca2+, Cu2+, Zn2+, and Ni2+ Acetivibrio thermocellus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.1 [(1->4)-alpha-D-glucosyl]n + phosphate Acetivibrio thermocellus
-
 [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.1 [(1->4)-alpha-D-glucosyl]n + phosphate Acetivibrio thermocellus ATCC 27405
-
 [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.1 Acetivibrio thermocellus
-
 ORF Cthe0357
-
2.4.1.1 Acetivibrio thermocellus ATCC 27405
-
 ORF Cthe0357
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.1 recombinant C-terminally His-tagged enzyme from Escherichia coli by nickel affinity chromatography, recombinant cellulose-binding module–intein-alphaGP fusion protein by affinity adsorption on amorphous cellulose followed by intein self-cleavage Acetivibrio thermocellus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.1 2.08
-
 purified recombinant detagged enzyme, pH 6.8, 50°C Acetivibrio thermocellus
2.4.1.1 2.22
-
 purified recombinant His-tagged enzyme, pH 6.8, 50°C Acetivibrio thermocellus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.1 dextrin DP13 + alpha-D-glucose 1-phosphate prolongation of the dextrin molecule with one glucose monomer Acetivibrio thermocellus dextrin DP14 + phosphate
-
 r
2.4.1.1 dextrin DP13 + alpha-D-glucose 1-phosphate prolongation of the dextrin molecule with one glucose monomer Acetivibrio thermocellus ATCC 27405 dextrin DP14 + phosphate
-
 r
2.4.1.1 dextrin DP14 + phosphate
-
 Acetivibrio thermocellus dextrin DP13 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.1 dextrin DP14 + phosphate
-
 Acetivibrio thermocellus ATCC 27405 dextrin DP13 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.1 dextrin DP18 + alpha-D-glucose 1-phosphate prolongation of the dextrin molecule with one glucose monomer Acetivibrio thermocellus dextrin DP19 + phosphate
-
 r
2.4.1.1 dextrin DP19 + phosphate
-
 Acetivibrio thermocellus dextrin DP18 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.1 dextrin DP3 + alpha-D-glucose 1-phosphate prolongation of the dextrin molecule with one glucose monomer Acetivibrio thermocellus dextrin DP4 + phosphate
-
 r
2.4.1.1 dextrin DP4 + phosphate
-
 Acetivibrio thermocellus dextrin DP3 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.1 additional information broad substrate specificities for different chain length dextrins and soluble starch, enzyme efficiency on dextrin DP4 is approximately 5fold lower than that on dextrin DP14 although kcat on dextrin DP4 is higher than on dextrin DP14 Acetivibrio thermocellus ?
-
 ?
2.4.1.1 additional information broad substrate specificities for different chain length dextrins and soluble starch, enzyme efficiency on dextrin DP4 is approximately 5fold lower than that on dextrin DP14 although kcat on dextrin DP4 is higher than on dextrin DP14 Acetivibrio thermocellus ATCC 27405 ?
-
 ?
2.4.1.1 starch + alpha-D-glucose 1-phosphate prolongation of the starch molecule with one glucose monomer Acetivibrio thermocellus starch + phosphate
-
 r
2.4.1.1 starch + alpha-D-glucose 1-phosphate prolongation of the starch molecule with one glucose monomer Acetivibrio thermocellus ATCC 27405 starch + phosphate
-
 r
2.4.1.1 [(1->4)-alpha-D-glucosyl]n + phosphate
-
 Acetivibrio thermocellus [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r
2.4.1.1 [(1->4)-alpha-D-glucosyl]n + phosphate
-
 Acetivibrio thermocellus ATCC 27405 [(1->4)-alpha-D-glucosyl]n-1 + alpha-D-glucose 1-phosphate
-
 r

Synonyms

EC Number Synonyms Comment Organism
2.4.1.1 alpha-glucan phosphorylase
-
 Acetivibrio thermocellus
2.4.1.1 alphaGP
-
 Acetivibrio thermocellus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.1 60
-
 synthesis direction Acetivibrio thermocellus
2.4.1.1 80
-
 degradation direction Acetivibrio thermocellus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.1.1 20 90 activity range Acetivibrio thermocellus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.1 40
-
 purified recombinant enzyme, loss of 10% activity within 120 min, stable Acetivibrio thermocellus
2.4.1.1 50
-
 half-life without additives is about 2 h, and is extended by 45fold to 90 h through a combined addition of 0.1 mM Mg2+, 5 mM DTT, 1% NaCl, 0.1% Triton X-100, and 1 mg/mL BSA Acetivibrio thermocellus
2.4.1.1 70
-
 purified recombinant enzyme, half-ife is about 40 min, inactivation after 120 min Acetivibrio thermocellus
2.4.1.1 80
-
 purified recombinant enzyme, loss of over 90% activity within 10 min, inactivation after 60 min Acetivibrio thermocellus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.1 2.1
-
 starch pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 3.6
-
 dextrin DP14 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 4.2
-
 alpha-D-glucose 1-phosphate pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 6.3
-
 starch pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 6.6
-
 dextrin DP19 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 8.1
-
 dextrin DP19 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 8.2
-
 phosphate pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 8.4
-
 dextrin DP4 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 9.3
-
 dextrin DP14 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 11
-
 dextrin DP4 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.1 6 6.5 synthesis direction Acetivibrio thermocellus
2.4.1.1 7 7.5 degradation direction Acetivibrio thermocellus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.1 4 10 activity range Acetivibrio thermocellus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.1.1 0.64
-
 dextrin DP4 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 1.1
-
 phosphate pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 1.5
-
 starch pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 3
-
 dextrin DP14 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 3.5
-
 dextrin DP19 pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 4.7
-
 dextrin DP4 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 7.7
-
 alpha-D-glucose 1-phosphate pH 6.8, 50°C, synthesis reaction Acetivibrio thermocellus
2.4.1.1 13
-
 dextrin DP14 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 21
-
 starch pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus
2.4.1.1 21
-
 dextrin DP19 pH 7.2, 50°C, degradation reaction Acetivibrio thermocellus