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Literature summary for 3.2.1.192 extracted from
- Enzymatic transformation of ginsenoside Rb1 to ginsenoside 20(S)-Rg3 by GH3 beta-glucosidase from Thermotoga thermarum DSM 5069 T (2015), J. Mol. Catal. B, 113, 104-109 .
No PubMed abstract available
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| Al3+ | 20% activation at 1 mM | Pseudothermotoga thermarum |  |
| Mn2+ | 41% activation at 1 mM | Pseudothermotoga thermarum | |
| additional information | no effect by 10 mM EDTA | Pseudothermotoga thermarum |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene Theth_1706, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain JM109(DE3) | Pseudothermotoga thermarum |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| Ag+ | 81% inhibition at 1 mM | Pseudothermotoga thermarum | |
| Cu2+ | 20% inhibition at 1 mM | Pseudothermotoga thermarum | |
| ethanol | inhibits 88% at 20% v/v, 30% at 10% v/v | Pseudothermotoga thermarum | |
| Fe2+ | 23% inhibition at 1 mM | Pseudothermotoga thermarum | |
| Hg2+ | 94% inhibition at 1 mM | Pseudothermotoga thermarum | |
| methanol | inhibits 94% at 20% v/v, 12% at 10% v/v | Pseudothermotoga thermarum | |
| additional information | no effect by 10 mM EDTA | Pseudothermotoga thermarum |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.065 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 5.0, 90°C, recombinant enzyme | Pseudothermotoga thermarum | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| additional information | no or poor effects by 1 mM of Ni2+, Fe3+, Mg2+, Zn2+, K+, Ca2+, Li+, and Co2+ | Pseudothermotoga thermarum |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ginsenoside Rb1 + H2O | Pseudothermotoga thermarum | - |
ginsenoside Rd + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | Pseudothermotoga thermarum | direct conversion of Rb1 into Rg3 | ginsenoside Rg3 + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | Pseudothermotoga thermarum DSM 5069 T | - |
ginsenoside Rd + D-glucopyranose | - |
? | |
| ginsenoside Rd + H2O | Pseudothermotoga thermarum | - |
ginsenoside Rg3 + D-glucopyranose | - |
? | |
| ginsenoside Rd + H2O | Pseudothermotoga thermarum DSM 5069 T | - |
ginsenoside Rg3 + D-glucopyranose | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudothermotoga thermarum | F7YVT1 | - |
- |
| Pseudothermotoga thermarum DSM 5069 T | F7YVT1 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme 2.9fold from Escherichia coli strain BL21(DE3) by heat treatment and nickel affinity chromatography | Pseudothermotoga thermarum |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
at a concentration of 0.75 U/mL of the enzyme, a temperature of 85°C and pH 5.0,10 g/l ginsenoside Rb1 is transformed into 6.9 g/l Rg3 within 60 min with a corresponding molar conversion of 97.8% | Pseudothermotoga thermarum |
| 70.7 | - |
purified recombinant His-tagged enzyme, pH 5.0, 90°C, substrate is 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Pseudothermotoga thermarum | 4-nitrophenol + beta-D-glucopyranose | - |
? | |
| 4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Pseudothermotoga thermarum DSM 5069 T | 4-nitrophenol + beta-D-glucopyranose | - |
? | |
| ginsenoside compound K + H2O | 33% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum | 20(S)-protopanaxadiol + D-glucopyranose | - |
? | |
| ginsenoside F2 + H2O | 34% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum | ginsenoside 20(S)-Rh2 + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | - |
Pseudothermotoga thermarum | ginsenoside Rd + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | 118% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum | ginsenoside Rd + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | - |
Pseudothermotoga thermarum DSM 5069 T | ginsenoside Rd + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | 118% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum DSM 5069 T | ginsenoside Rd + D-glucopyranose | - |
? | |
| ginsenoside Rb1 + H2O | direct conversion of Rb1 into Rg3 | Pseudothermotoga thermarum | ginsenoside Rg3 + D-glucopyranose | - |
? | |
| ginsenoside Rd + H2O | - |
Pseudothermotoga thermarum | ginsenoside Rg3 + D-glucopyranose | - |
? | |
| ginsenoside Rd + H2O | - |
Pseudothermotoga thermarum DSM 5069 T | ginsenoside Rg3 + D-glucopyranose | - |
? | |
| ginsenoside Rd + H2O | 43% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum | ginsenoside 20(S)-Rg3 + D-glucopyranose | - |
? | |
| ginsenoside Re + H2O | 110% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum | ginsenoside Rg2 + D-glucopyranose | - |
? | |
| ginsenoside Rg1 + H2O | 81% activity compared to 4-nitrophenyl beta-D-glucopyranoside | Pseudothermotoga thermarum | ginsenoside 20(S)-Rh1 + D-glucopyranose | - |
? | |
| additional information | the beta-glucosidase exhibits high selectivity to cleave the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1 and catalyzes the conversion of ginsenoside Rb1 or Rd to the more pharmacologically active minor ginsenoside 20(S)-Rg3. The enzyme is also able to hydrolyze cellobiose and gentiobiose. No activity is detected with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-beta-D-xylopyranoside, 4-nitrophenyl-alpha-L-arabinofuranoside, carboxymethyl cellulose, and sucrose | Pseudothermotoga thermarum | ? | - |
? | |
| additional information | the beta-glucosidase exhibits high selectivity to cleave the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1 and catalyzes the conversion of ginsenoside Rb1 or Rd to the more pharmacologically active minor ginsenoside 20(S)-Rg3. The enzyme is also able to hydrolyze cellobiose and gentiobiose. No activity is detected with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-beta-D-xylopyranoside, 4-nitrophenyl-alpha-L-arabinofuranoside, carboxymethyl cellulose, and sucrose | Pseudothermotoga thermarum DSM 5069 T | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 82000, recombinant His-tagged enzyme, SDS-PAGE | Pseudothermotoga thermarum |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| BGL3T | - |
Pseudothermotoga thermarum |
| GH3 beta-glucosidase | - |
Pseudothermotoga thermarum |
| More | cf. EC 3.2.1.21 | Pseudothermotoga thermarum |
| Theth_1706 | - |
Pseudothermotoga thermarum |
| Tt-BGL | - |
Pseudothermotoga thermarum |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
recombinant enzyme | Pseudothermotoga thermarum |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 65 | 100 | activity profile of the recombinant enzyme, overview | Pseudothermotoga thermarum |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 85 | - |
purified recombinant His-tagged enzyme, pH 5.0, about 90% activity remaining after 120 min | Pseudothermotoga thermarum |
| 90 | - |
purified recombinant His-tagged enzyme, pH 5.0, 50% activity remaining after 60 min, 40% after 120 min | Pseudothermotoga thermarum |
| 95 | - |
purified recombinant His-tagged enzyme, pH 5.0, about 10% activity remaining after 120 min, 20% after 60 min | Pseudothermotoga thermarum |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 121 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 5.0, 90°C, recombinant enzyme | Pseudothermotoga thermarum | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5 | - |
recombinant enzyme | Pseudothermotoga thermarum |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4.5 | 6.5 | over 50% of maximal activity within this range | Pseudothermotoga thermarum |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 4.5 | 7.5 | purified recombinant His-tagged enzyme, 80% activity remaining at pH 4.5-7.5 | Pseudothermotoga thermarum |
| 5.5 | - |
purified recombinant His-tagged enzyme, completely stable at | Pseudothermotoga thermarum |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the enzyme belongs to the glycosyl hydrolase family 3, GH3 | Pseudothermotoga thermarum |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1861 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 5.0, 90°C, recombinant enzyme | Pseudothermotoga thermarum | |
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