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Literature summary extracted from

  • Nam, H.K.; Hong, S.H.; Shin, K.C.; Oh, D.K.
    Quercetin production from rutin by a thermostable beta-rutinosidase from Pyrococcus furiosus (2012), Biotechnol. Lett., 34, 483-489.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.B31 1.4-dioxane
-
 Pyrococcus furiosus
3.2.1.B31 2-propanol
-
 Pyrococcus furiosus
3.2.1.B31 3-methyl-1-butanol
-
 Pyrococcus furiosus
3.2.1.B31 acetone
-
 Pyrococcus furiosus
3.2.1.B31 DMSO
-
 Pyrococcus furiosus
3.2.1.B31 ethanol
-
 Pyrococcus furiosus
3.2.1.B31 Isopropyl alcohol
-
 Pyrococcus furiosus
3.2.1.B31 methanol
-
 Pyrococcus furiosus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.B28
-
 Pyrococcus furiosus
3.2.1.B31
-
 Pyrococcus furiosus
3.2.1.B31 DNA aand amino acid sequence determination and analysis, expression in Escherichia coli strain ER2566, the enzyme is secreted Pyrococcus furiosus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.B28 0.17
-
 isoquercitrin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B28 3.095
-
 quercitrin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.07
-
 quercitrin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 0.17
-
 isoquercitrin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 0.34
-
 rutin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.34
-
 rutin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.B31 isoquercitrin + H2O Pyrococcus furiosus best substrate quercetin + beta-D-glucose
-
 ?
3.2.1.B31 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 ?
-
 ?
3.2.1.B31 quercitrin + H2O Pyrococcus furiosus
-
 quercetin + L-rhamnose
-
 ?
3.2.1.B31 rutin + H2O Pyrococcus furiosus
-
 quercetin + rutinose
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.B28 Pyrococcus furiosus Q51723
-
-
3.2.1.B31 Pyrococcus furiosus Q51723
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.B31 recombinant extracellular enzyme from culture supernatant by heat treatment and centrifugation Pyrococcus furiosus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.B28 0.23
-
 substrate: quercitrin, pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B28 220
-
 substrate: isoquercitrin, pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.03
-
 substrate: rutin, pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.03
-
 substrate rutin, pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 0.23
-
 substrate quercitrin, pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 220
-
 substrate isoquercitrin, pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.B28 isoquercitrin + H2O i.e. quercetin 3-O-beta-D-glucoside Pyrococcus furiosus quercetin + beta-D-glucose
-
 ?
3.2.1.B28 additional information the enzyme shows also beta-rutinosidase activity with rutin (hydrolysis of rutin to quercetin and rutinose) Pyrococcus furiosus ?
-
 ?
3.2.1.B28 quercitrin + H2O i.e. quercetin 3-O-alpha-L-rhamnoside Pyrococcus furiosus quercetin + alpha-L-rhamnose
-
 ?
3.2.1.B31 2-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 2-nitrophenol + D-glucose
-
 ?
3.2.1.B31 4-nitrophenyl beta-D-glucopyranoside + H2O
-
 Pyrococcus furiosus 4-nitrophenol + D-glucose
-
 ?
3.2.1.B31 isoquercitrin + H2O best substrate Pyrococcus furiosus quercetin + beta-D-glucose
-
 ?
3.2.1.B31 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 ?
-
 ?
3.2.1.B31 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 ?
-
 ?
3.2.1.B31 quercitrin + H2O
-
 Pyrococcus furiosus quercetin + L-rhamnose
-
 ?
3.2.1.B31 rutin + H2O
-
 Pyrococcus furiosus quercetin + rutinose
-
 ?
3.2.1.B31 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 ?
3.2.1.B31 rutin + H2O i.e. quercetin 3-O-beta-D-rutenoside Pyrococcus furiosus quercetin + rutinose
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.B28 95
-
-
 Pyrococcus furiosus
3.2.1.B31 95
-
-
 Pyrococcus furiosus
3.2.1.B31 95
-
 assay at Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.B28 75
-
 half-life: 1017 h Pyrococcus furiosus
3.2.1.B28 80
-
 half-life: 461 h Pyrococcus furiosus
3.2.1.B28 85
-
 half-life: 286 h Pyrococcus furiosus
3.2.1.B28 90
-
 half-life: 173 h Pyrococcus furiosus
3.2.1.B28 95
-
 half-life: 101 h Pyrococcus furiosus
3.2.1.B31 75
-
 half-life: 1017 h Pyrococcus furiosus
3.2.1.B31 80
-
 half-life: 461 h Pyrococcus furiosus
3.2.1.B31 85
-
 half-life: 286 h Pyrococcus furiosus
3.2.1.B31 90
-
 half-life: 173 h Pyrococcus furiosus
3.2.1.B31 95
-
 half-life: 101 h Pyrococcus furiosus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.B28 0.22
-
 quercitrin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B28 53.3
-
 isoquercitrin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.026
-
 rutin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 0.027
-
 rutin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.22
-
 quercitrin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 53.3
-
 isoquercitrin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.B28 5
-
-
 Pyrococcus furiosus
3.2.1.B31 5
-
-
 Pyrococcus furiosus
3.2.1.B31 5
-
 assay at Pyrococcus furiosus

General Information

EC Number General Information Comment Organism
3.2.1.B31 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.21 2.33
-
 quercitrin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.21 303
-
 isoquercitrin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 0.075
-
 rutin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 0.078
-
 rutin pH 5.0, 95°C Pyrococcus furiosus
3.2.1.B31 2.33
-
 quercitrin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus
3.2.1.B31 303.3
-
 isoquercitrin pH 5.0, 09°C, recombinant enzyme Pyrococcus furiosus