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Literature summary extracted from
- Purification method improvement and characterization of a novel ginsenoside-hydrolyzing beta-glucosidase from Paecilomyces Bainier sp. 229 (2008), Biosci. Biotechnol. Biochem., 72, 352-359.
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 3.2.1.21 | 0.072 | - |
ginsenoside Rb1 | pH 3.5, 45°C | Paecilomyces sp. (in: Eurotiomycetes) |  |
| 3.2.1.21 | 0.111 | - |
4-nitrophenyl-beta-D-glucopyranoside | pH 3.5, 45°C | Paecilomyces sp. (in: Eurotiomycetes) | |
| 3.2.1.21 | 0.805 | - |
ginsenoside Rd | pH 3.5, 45°C | Paecilomyces sp. (in: Eurotiomycetes) | |
| 3.2.1.192 | 0.072 | - |
ginsenoside Rb1 | pH 3.5, 45°C | Paecilomyces sp. (in: Eurotiomycetes) | |
| 3.2.1.192 | 0.111 | - |
4-nitrophenyl beta-D-glucoside | pH 3.5, 45°C | Paecilomyces sp. (in: Eurotiomycetes) | |
| 3.2.1.192 | 0.805 | - |
ginsenoside Rd | pH 3.5, 45°C | Paecilomyces sp. (in: Eurotiomycetes) | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 102000 | - |
3 * 102000, SDS-PAGE | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.21 | 305000 | - |
gel filtration | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 102000 | - |
3 * 102000, SDS-PAGE | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 305000 | - |
gel filtration | Paecilomyces sp. (in: Eurotiomycetes) |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 3.2.1.21 | Paecilomyces sp. (in: Eurotiomycetes) | - |
- |
- |
| 3.2.1.21 | Paecilomyces sp. (in: Eurotiomycetes) 229 | - |
- |
- |
| 3.2.1.192 | Paecilomyces sp. (in: Eurotiomycetes) | - |
- |
- |
| 3.2.1.192 | Paecilomyces sp. (in: Eurotiomycetes) 220 Bainier | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 3.2.1.21 | native enzyme 144fold by ammonium sulfate fractionation, anion exchange chromatography, and affinity chromatography on microcrystalline cellulose, to homogeneity, method optimization, overview | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | only two chromatographic steps, QSepharose FF and microcrystalline cellulose column in sequence, are required to purify the enzyme to homogeneity | Paecilomyces sp. (in: Eurotiomycetes) |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 3.2.1.192 | ginsenoside Rb1 + 2 H2O = ginsenoside Rg3 + 2 D-glucopyranose | overall reaction | Paecilomyces sp. (in: Eurotiomycetes) |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 56.7 | - |
purified enzyme, substrate ginsenoside Rb1 | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.21 | 129.4 | - |
purified enzyme, substrate ginsenoside Rd | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.21 | 170.5 | - |
purified enzyme, substrate 4-nitrophenyl-beta-D-glucopyranoside | Paecilomyces sp. (in: Eurotiomycetes) |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 3.2.1.21 | 4-nitrophenyl beta-D-glucopyranoside + H2O | best substrate | Paecilomyces sp. (in: Eurotiomycetes) | 4-nitrophenol + beta-D-glucose | - |
? | |
| 3.2.1.21 | 4-nitrophenyl beta-D-glucopyranoside + H2O | best substrate | Paecilomyces sp. (in: Eurotiomycetes) 229 | 4-nitrophenol + beta-D-glucose | - |
? | |
| 3.2.1.21 | 4-nitrophenyl beta-D-glucopyranoside + H2O | best substrate | Paecilomyces sp. (in: Eurotiomycetes) 220 Bainier | 4-nitrophenol + beta-D-glucose | - |
? | |
| 3.2.1.21 | cellobiose + H2O | low activity | Paecilomyces sp. (in: Eurotiomycetes) | beta-D-glucose + alpha-D-glucose | - |
? | |
| 3.2.1.21 | cellobiose + H2O | low activity | Paecilomyces sp. (in: Eurotiomycetes) 229 | beta-D-glucose + alpha-D-glucose | - |
? | |
| 3.2.1.21 | cellobiose + H2O | low activity | Paecilomyces sp. (in: Eurotiomycetes) 220 Bainier | beta-D-glucose + alpha-D-glucose | - |
? | |
| 3.2.1.21 | gentiobiose + H2O | - |
Paecilomyces sp. (in: Eurotiomycetes) | 2 beta-D-glucose | - |
? | |
| 3.2.1.21 | gentiobiose + H2O | - |
Paecilomyces sp. (in: Eurotiomycetes) 229 | 2 beta-D-glucose | - |
? | |
| 3.2.1.21 | gentiobiose + H2O | - |
Paecilomyces sp. (in: Eurotiomycetes) 220 Bainier | 2 beta-D-glucose | - |
? | |
| 3.2.1.21 | ginsenoside Rb1 + H2O | - |
Paecilomyces sp. (in: Eurotiomycetes) | ? | - |
? | |
| 3.2.1.21 | ginsenoside Rd + H2O | - |
Paecilomyces sp. (in: Eurotiomycetes) | ? | - |
? | |
| 3.2.1.21 | additional information | maltose is a poor substrate, no activity with ginsenoside Rg3, Rb2, and Rb3, and with 4-nitrophenyl-alpha-D-glucopyranoside and 4-nitrophenyl-beta-D-xylopyranoside | Paecilomyces sp. (in: Eurotiomycetes) | ? | - |
? | |
| 3.2.1.21 | additional information | maltose is a poor substrate, no activity with ginsenoside Rg3, Rb2, and Rb3, and with 4-nitrophenyl-alpha-D-glucopyranoside and 4-nitrophenyl-beta-D-xylopyranoside | Paecilomyces sp. (in: Eurotiomycetes) 229 | ? | - |
? | |
| 3.2.1.21 | additional information | maltose is a poor substrate, no activity with ginsenoside Rg3, Rb2, and Rb3, and with 4-nitrophenyl-alpha-D-glucopyranoside and 4-nitrophenyl-beta-D-xylopyranoside | Paecilomyces sp. (in: Eurotiomycetes) 220 Bainier | ? | - |
? | |
| 3.2.1.21 | sophorose + H2O | low activity | Paecilomyces sp. (in: Eurotiomycetes) | 2 beta-D-glucose | - |
? | |
| 3.2.1.21 | sophorose + H2O | low activity | Paecilomyces sp. (in: Eurotiomycetes) 229 | 2 beta-D-glucose | - |
? | |
| 3.2.1.21 | sophorose + H2O | low activity | Paecilomyces sp. (in: Eurotiomycetes) 220 Bainier | 2 beta-D-glucose | - |
? | |
| 3.2.1.192 | 4-nitrophenyl beta-D-glucoside + H2O | no hydrolysis of 4-nitrophenyl-alpha-D-glucoside or 4-nitrophenyl-beta-D-xyloside | Paecilomyces sp. (in: Eurotiomycetes) | 4-nitrophenol + D-glucopyranose | - |
? | |
| 3.2.1.192 | gentiobiose + H2O | - |
Paecilomyces sp. (in: Eurotiomycetes) | D-glucopyranose | - |
? | |
| 3.2.1.192 | ginsenoside Rb1 + H2O | hydrolysis of 20-O-beta-D-glucosyl-beta-D-(1->6)-glucosidic bond | Paecilomyces sp. (in: Eurotiomycetes) | ginsenoside Rd + D-glucopyranose | - |
? | |
| 3.2.1.192 | ginsenoside Rd + H2O | hydrolyzes the 20-O-glucoside of Rd at 44% of the activity compared to the hydrolysis of the 20-O-beta-D-glucosyl-beta-D-(1->6)-glucosidic bond of ginsenoside Rb1 | Paecilomyces sp. (in: Eurotiomycetes) | ginsenoside Rg3 + D-glucopyranose | - |
? | |
| 3.2.1.192 | additional information | no hydrolysis of the 20-O-alpha-D-(1->6)-arabinopyranosidic bond of ginsenoside Rb2 and the 20-O-beta-D-(1->6)-xylosidic bond of ginsenoside Rb3. It does not hydrolyse the 3-O-beta-glucosidic bond of ginsenoside Rb1, Rb2, Rb3 | Paecilomyces sp. (in: Eurotiomycetes) | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 3.2.1.21 | trimer | 3 * 102000, SDS-PAGE | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | trimer | 3 * 102000, SDS-PAGE | Paecilomyces sp. (in: Eurotiomycetes) |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 55 | - |
- |
Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 45 | - |
assay at | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 55 | - |
- |
Paecilomyces sp. (in: Eurotiomycetes) |
Temperature Range [°C]
| EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.192 | 40 | 60 | 40°C: about 50% of maximal activity. 60°C: about 60% of maximal activity | Paecilomyces sp. (in: Eurotiomycetes) |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 55 | - |
purified enzyme, 4 h, pH 3.5, stable up to, rapid decrease in activity above | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 50 | - |
pH 3.5, 4 h, 10.4% loss of activity | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 55 | - |
pH 3.5, 4 h, 81.5% loss of activity | Paecilomyces sp. (in: Eurotiomycetes) |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 3.5 | - |
- |
Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 3.5 | - |
at 55°C, in 50 mM formate buffer | Paecilomyces sp. (in: Eurotiomycetes) |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 3 | 4 | rapid decrease in activity below pH 3.0 and above pH 4.0 | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 2.5 | 5 | pH 2.5: about 45% of maximal activity, pH 5.0: about 45% of maximal activity | Paecilomyces sp. (in: Eurotiomycetes) |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 3.2.1.21 | 3 | 7 | purified enzyme, 4 h, 45°C, stable | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 3 | 7 | 45°C, 4 h, 10% loss of activity | Paecilomyces sp. (in: Eurotiomycetes) |
| 3.2.1.192 | 3.5 | 5 | 45°C, 4 h, 5% loss of activity | Paecilomyces sp. (in: Eurotiomycetes) |
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