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Literature summary extracted from

  • Silva, Z.; Alarico, S.; da Costa, M.S.
    Trehalose biosynthesis in Thermus thermophilus RQ-1: biochemical properties of the trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase (2005), Extremophiles, 9, 29-36.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.1.3.12 ADP-glucose activates 4.5% at 5 mM Thermus thermophilus
3.1.3.12 GDP-glucose activates 3.6% at 5 mM Thermus thermophilus
3.1.3.12 TDP-glucose activates 4.6% at 5 mM Thermus thermophilus
3.1.3.12 UDP-glucose activates 10% at 5 mM Thermus thermophilus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.15 gene otsA, cloned from genetic library, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain BL21 Thermus thermophilus
3.1.3.12 gene otsB, DNA and amino acid sequence determination and analysis, overexpression of GST-tagged enzyme in Eschrichia coli strains XL1-Blue and BL21 Thermus thermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.3.12 Ba2+
-
 Thermus thermophilus
3.1.3.12 Ca2+
-
 Thermus thermophilus
3.1.3.12 Cu2+
-
 Thermus thermophilus
3.1.3.12 Sr2+
-
 Thermus thermophilus
3.1.3.12 Zn2+
-
 Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.15 0.5
-
 UDP-glucose recombinant enzyme, pH 7.0, 80°C, in presence of Mg2+ Thermus thermophilus
2.4.1.15 0.8
-
 D-glucose 6-phosphate recombinant enzyme, pH 7.0, 80°C, in presence of Mg2+ Thermus thermophilus
3.1.3.12 1.5
-
 trehalose 6-phosphate pH 7.0, 70°C, recombinant enzyme Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.15 Co2+ stimulates Thermus thermophilus
2.4.1.15 Mg2+ stimulates Thermus thermophilus
2.4.1.15 additional information no requirement for divalent cations for activity Thermus thermophilus
3.1.3.12 Co2+ stimulates Thermus thermophilus
3.1.3.12 Mg2+ stimulates Thermus thermophilus
3.1.3.12 additional information no effect by Mn2+, the enzyme requires divalent cations for maximal activity, in absence of metal ion the enzyme shows 32.5% of maximal activity Thermus thermophilus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.15 52000
-
 x * 52000, recombinant enzyme, SDS-PAGE Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.15 UDP-alpha-D-glucose + D-glucose 6-phosphate Thermus thermophilus first step in one of 2 existing trehalose biosynthesis pathways, plays a role in osmoadaptation UDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 UDP-alpha-D-glucose + D-glucose 6-phosphate Thermus thermophilus RQ-1 first step in one of 2 existing trehalose biosynthesis pathways, plays a role in osmoadaptation UDP + alpha,alpha-trehalose 6-phosphate
-
 ?
3.1.3.12 trehalose 6-phosphate + H2O Thermus thermophilus the trehalose pathway involving trehalose-6-phosphate phosphatase plays a role in osmoadaptation, enzyme deficiency leads to reduced osmotolerance trehalose + phosphate
-
 ?
3.1.3.12 trehalose 6-phosphate + H2O Thermus thermophilus RQ-1 the trehalose pathway involving trehalose-6-phosphate phosphatase plays a role in osmoadaptation, enzyme deficiency leads to reduced osmotolerance trehalose + phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.15 Thermus thermophilus Q7WUI7 gene otsA
-
2.4.1.15 Thermus thermophilus RQ-1 Q7WUI7 gene otsA
-
3.1.3.12 Thermus thermophilus Q7WUI6 gene otsB
-
3.1.3.12 Thermus thermophilus RQ-1 Q7WUI6 gene otsB
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.15 recombinant enzyme from Escherichia coli Thermus thermophilus
3.1.3.12 recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography, gel filtration, and ion exchange chromatography, to homogeneity, the GST-tag is cleaved off by thrombin Thermus thermophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.15 20.6
-
 purified recombinant enzyme, substrates GDP-glucose and D-glucose 6-phosphate Thermus thermophilus
2.4.1.15 95
-
 purified recombinant enzyme, substrates ADP-glucose and D-glucose 6-phosphate Thermus thermophilus
2.4.1.15 106.7
-
 purified recombinant enzyme, substrates TDP-glucose and D-glucose 6-phosphate Thermus thermophilus
2.4.1.15 149.6
-
 purified recombinant enzyme, substrates UDP-glucose and D-glucose 6-phosphate Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.15 ADP-alpha-D-glucose + D-glucose 6-phosphate less effective than UDP-glucose Thermus thermophilus ADP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 ADP-alpha-D-glucose + D-glucose 6-phosphate less effective than UDP-glucose Thermus thermophilus RQ-1 ADP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 GDP-alpha-D-glucose + D-glucose 6-phosphate much less effective than UDP-glucose Thermus thermophilus GDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 GDP-alpha-D-glucose + D-glucose 6-phosphate much less effective than UDP-glucose Thermus thermophilus RQ-1 GDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 additional information donor substrate specificity in descending order: UDP-glucose, TDP-glucose, ADP-glucose, GDP-glucose, no activity with mannose-6-phosphate, fructose 6-phosphate, or glucose 1,6-diphosphate as acceptors Thermus thermophilus ?
-
 ?
2.4.1.15 additional information donor substrate specificity in descending order: UDP-glucose, TDP-glucose, ADP-glucose, GDP-glucose, no activity with mannose-6-phosphate, fructose 6-phosphate, or glucose 1,6-diphosphate as acceptors Thermus thermophilus RQ-1 ?
-
 ?
2.4.1.15 TDP-alpha-D-glucose + D-glucose 6-phosphate less effective than UDP-glucose Thermus thermophilus TDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 UDP-alpha-D-glucose + D-glucose 6-phosphate first step in one of 2 existing trehalose biosynthesis pathways, plays a role in osmoadaptation Thermus thermophilus UDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 UDP-alpha-D-glucose + D-glucose 6-phosphate best substrates, absolutely specific for D-glucose 6-phosphate Thermus thermophilus UDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 UDP-alpha-D-glucose + D-glucose 6-phosphate first step in one of 2 existing trehalose biosynthesis pathways, plays a role in osmoadaptation Thermus thermophilus RQ-1 UDP + alpha,alpha-trehalose 6-phosphate
-
 ?
2.4.1.15 UDP-alpha-D-glucose + D-glucose 6-phosphate best substrates, absolutely specific for D-glucose 6-phosphate Thermus thermophilus RQ-1 UDP + alpha,alpha-trehalose 6-phosphate
-
 ?
3.1.3.12 D-glucose 6-phosphate + H2O 3.1% of activity with trehalose 6-phosphate Thermus thermophilus D-glucose + phosphate
-
 ?
3.1.3.12 D-glucose 6-phosphate + H2O 3.1% of activity with trehalose 6-phosphate Thermus thermophilus RQ-1 D-glucose + phosphate
-
 ?
3.1.3.12 additional information glucose diphosphate nucleosides are no substrates Thermus thermophilus ?
-
 ?
3.1.3.12 additional information glucose diphosphate nucleosides are no substrates Thermus thermophilus RQ-1 ?
-
 ?
3.1.3.12 trehalose 6-phosphate + H2O the trehalose pathway involving trehalose-6-phosphate phosphatase plays a role in osmoadaptation, enzyme deficiency leads to reduced osmotolerance Thermus thermophilus trehalose + phosphate
-
 ?
3.1.3.12 trehalose 6-phosphate + H2O high substrate specificity of recombinant enzyme Thermus thermophilus trehalose + phosphate
-
 ?
3.1.3.12 trehalose 6-phosphate + H2O the trehalose pathway involving trehalose-6-phosphate phosphatase plays a role in osmoadaptation, enzyme deficiency leads to reduced osmotolerance Thermus thermophilus RQ-1 trehalose + phosphate
-
 ?
3.1.3.12 trehalose 6-phosphate + H2O high substrate specificity of recombinant enzyme Thermus thermophilus RQ-1 trehalose + phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.4.1.15 ? x * 52000, recombinant enzyme, SDS-PAGE Thermus thermophilus

Synonyms

EC Number Synonyms Comment Organism
2.4.1.15 TPS
-
 Thermus thermophilus
2.4.1.15 trehalose-6-phosphate synthase
-
 Thermus thermophilus
3.1.3.12 TPP
-
 Thermus thermophilus
3.1.3.12 trehalose-6-phosphate phosphatase
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.15 98
-
-
 Thermus thermophilus
3.1.3.12 70
-
 recombinant enzyme Thermus thermophilus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.1.15 40 100 about half-maximal activity at 40°C, 80% of maximal activity at 100°C, inactive at 30°C Thermus thermophilus
3.1.3.12 40 90 recombinant enzyme, 45% of maximal activity at 90°C, about 15% at 40°C Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.15 80
-
 half-life: 193 min Thermus thermophilus
2.4.1.15 90
-
 half-life: 54 min Thermus thermophilus
2.4.1.15 100
-
 half-life: 31 min Thermus thermophilus
3.1.3.12 80
-
 half-life of the recombinant enzyme is 161 min Thermus thermophilus
3.1.3.12 90
-
 half-life of the recombinant enzyme is 120 min Thermus thermophilus
3.1.3.12 100
-
 half-life of the recombinant enzyme is 40 min Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.15 6
-
-
 Thermus thermophilus
3.1.3.12 7
-
 about, recombinant enzyme Thermus thermophilus