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Literature summary for 3.2.1.B31 extracted from
- Quercetin production from rutin by a thermostable beta-rutinosidase from Pyrococcus furiosus (2012), Biotechnol. Lett., 34, 483-489.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 1.4-dioxane | - |
Pyrococcus furiosus |  |
| 2-propanol | - |
Pyrococcus furiosus | |
| 3-methyl-1-butanol | - |
Pyrococcus furiosus | |
| acetone | - |
Pyrococcus furiosus | |
| DMSO | - |
Pyrococcus furiosus | |
| ethanol | - |
Pyrococcus furiosus | |
| Isopropyl alcohol | - |
Pyrococcus furiosus | |
| methanol | - |
Pyrococcus furiosus | |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
- |
Pyrococcus furiosus |
| DNA aand amino acid sequence determination and analysis, expression in Escherichia coli strain ER2566, the enzyme is secreted | Pyrococcus furiosus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.07 | - |
quercitrin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
| 0.17 | - |
isoquercitrin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
| 0.34 | - |
rutin | pH 5.0, 95°C | Pyrococcus furiosus | |
| 0.34 | - |
rutin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| isoquercitrin + H2O | Pyrococcus furiosus | best substrate | quercetin + beta-D-glucose | - |
? | |
| additional information | Pyrococcus furiosus | no or poor activity with 4-nitrophenyl alpha-D-glucopyranoside and 4-nitrophenyl alpha-L-rhamnopyranoside, no activity with rutinose, maltose, and turanose | ? | - |
? | |
| quercitrin + H2O | Pyrococcus furiosus | - |
quercetin + L-rhamnose | - |
? | |
| rutin + H2O | Pyrococcus furiosus | - |
quercetin + rutinose | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pyrococcus furiosus | Q51723 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant extracellular enzyme from culture supernatant by heat treatment and centrifugation | Pyrococcus furiosus |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 0.03 | - |
substrate: rutin, pH 5.0, 95°C | Pyrococcus furiosus |
| 0.03 | - |
substrate rutin, pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus |
| 0.23 | - |
substrate quercitrin, pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus |
| 220 | - |
substrate isoquercitrin, pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-nitrophenyl beta-D-glucopyranoside + H2O | - |
Pyrococcus furiosus | 2-nitrophenol + D-glucose | - |
? | |
| 4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Pyrococcus furiosus | 4-nitrophenol + D-glucose | - |
? | |
| isoquercitrin + H2O | best substrate | Pyrococcus furiosus | quercetin + beta-D-glucose | - |
? | |
| additional information | the enzyme shows also beta-glucosidase activity with isoquercitrin (i.e. quercetin 3-O-beta-D-glucoside) or quercitrin (i.e. quercetin 3-O-alpha-L-rhamnoside) | Pyrococcus furiosus | ? | - |
? | |
| additional information | no or poor activity with 4-nitrophenyl alpha-D-glucopyranoside and 4-nitrophenyl alpha-L-rhamnopyranoside, no activity with rutinose, maltose, and turanose | Pyrococcus furiosus | ? | - |
? | |
| quercitrin + H2O | - |
Pyrococcus furiosus | quercetin + L-rhamnose | - |
? | |
| rutin + H2O | - |
Pyrococcus furiosus | quercetin + rutinose | - |
? | |
| rutin + H2O | - |
Pyrococcus furiosus | quercetin + rutinose | formation in of quercetin and rutinose disaccharide with a ratio of 1:1, with no glucose, L-rhamnose, and isoquercitrin | ? | |
| rutin + H2O | i.e. quercetin 3-O-beta-D-rutenoside | Pyrococcus furiosus | quercetin + rutinose | - |
? | |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 95 | - |
- |
Pyrococcus furiosus |
| 95 | - |
assay at | Pyrococcus furiosus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 75 | - |
half-life: 1017 h | Pyrococcus furiosus |
| 80 | - |
half-life: 461 h | Pyrococcus furiosus |
| 85 | - |
half-life: 286 h | Pyrococcus furiosus |
| 90 | - |
half-life: 173 h | Pyrococcus furiosus |
| 95 | - |
half-life: 101 h | Pyrococcus furiosus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.026 | - |
rutin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
| 0.027 | - |
rutin | pH 5.0, 95°C | Pyrococcus furiosus | |
| 0.22 | - |
quercitrin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
| 53.3 | - |
isoquercitrin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 5 | - |
- |
Pyrococcus furiosus |
| 5 | - |
assay at | Pyrococcus furiosus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | homology modelling using crystal structure of Thermosphaera aggregans beta-glucosidase, PDB ID 1QVB, molecular dynamics, molecular docking of rutin or isoquercitrin as a substrate into the predicted active site, overview. An additional L-rhamnose in rutin might disrupt the binding of rutin to the catalytic residues because of its bulky size and hydroxyl groups | Pyrococcus furiosus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.075 | - |
rutin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
| 0.078 | - |
rutin | pH 5.0, 95°C | Pyrococcus furiosus | |
| 2.33 | - |
quercitrin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
| 303.3 | - |
isoquercitrin | pH 5.0, 09°C, recombinant enzyme | Pyrococcus furiosus | |
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