Activating Compound | Comment | Organism | Structure |
---|---|---|---|
methanol | activates 14% at 5%, inhibits 20% at 30% | Thermotoga petrophila |
Cloned (Comment) | Organism |
---|---|
gene Tpet_0898, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) partly in inclusion bodies, method optimization | Thermotoga petrophila |
General Stability | Organism |
---|---|
Ca2+ and Mn2+ stabilize the purified recombinant enzyme | Thermotoga petrophila |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Al3+ | 67% inhibition at 5 mM | Thermotoga petrophila | |
Cu2+ | 40% inhibition at 5 mM | Thermotoga petrophila | |
DMSO | inhibits 26% at 15% and 42% at 30% | Thermotoga petrophila | |
ethanol | inhibits 20% at 15% and 45% at 30% | Thermotoga petrophila | |
Hg2+ | complete inhibition at 1-5 mM | Thermotoga petrophila | |
methanol | activates 14% at 5%, inhibits 20% at 30% | Thermotoga petrophila | |
additional information | no significant effect of 5 mM EDTA on the enzyme activity | Thermotoga petrophila | |
NH4+ | 45% inhibition at 5 mM | Thermotoga petrophila |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates about 1.5fold at 5 mM, stabilizes the purified recombinant enzyme | Thermotoga petrophila | |
Co2+ | activates about 1.5fold at 5 mM, does not stabilize the purified recombinant enzyme | Thermotoga petrophila | |
Mn2+ | activates about 1.5fold at 5 mM, stabilizes the purified recombinant enzyme | Thermotoga petrophila |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ginsenoside Rb1 + H2O | Thermotoga petrophila | - |
ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | Thermotoga petrophila | direct conversion of Rb1 into Rg3 | ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | Thermotoga petrophila DSM 13995 | - |
ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | Thermotoga petrophila RKU-1 | - |
ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | Thermotoga petrophila ATCC BAA-488 | - |
ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | Thermotoga petrophila | - |
ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | Thermotoga petrophila DSM 13995 | - |
ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | Thermotoga petrophila RKU-1 | - |
ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | Thermotoga petrophila ATCC BAA-488 | - |
ginsenoside Rg3 + D-glucopyranose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermotoga petrophila | A5IL43 | - |
- |
Thermotoga petrophila ATCC BAA-488 | A5IL43 | - |
- |
Thermotoga petrophila DSM 13995 | A5IL43 | - |
- |
Thermotoga petrophila RKU-1 | A5IL43 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme 6.1fold from Escherichia coli strain BL21(DE3) by heat treatment and nickel affinity chromatography | Thermotoga petrophila |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
in a reaction at 90°C and pH 5.0, 10 g/l of ginsenoside Rb1 is transformed into 6.93 g/l of Rg3 within 90 min, with a corresponding molar conversion of 97.9 %, and Rg3 productivity of 4620 mg/l/h | Thermotoga petrophila |
95.7 | - |
purified recombinant His-tagged enzyme, pH 5.0, 90°C, substrate is 4-nitrophenyl beta-D-glucopyranoside | Thermotoga petrophila |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-nitrophenyl beta-D-glucopyranoside + H2O | low activity | Thermotoga petrophila | 2-nitrophenol + beta-D-glucopyranose | - |
? | |
2-nitrophenyl beta-D-glucopyranoside + H2O | low activity | Thermotoga petrophila DSM 13995 | 2-nitrophenol + beta-D-glucopyranose | - |
? | |
2-nitrophenyl beta-D-glucopyranoside + H2O | low activity | Thermotoga petrophila RKU-1 | 2-nitrophenol + beta-D-glucopyranose | - |
? | |
2-nitrophenyl beta-D-glucopyranoside + H2O | low activity | Thermotoga petrophila ATCC BAA-488 | 2-nitrophenol + beta-D-glucopyranose | - |
? | |
4-nitrophenyl beta-D-glucopyranoside + H2O | - |
Thermotoga petrophila | 4-nitrophenol + beta-D-glucopyranose | - |
? | |
ginsenoside compound K + H2O | 35% activity compared to ginsenoside Rb1 | Thermotoga petrophila | 20(S)-protopanaxadiol + D-glucopyranose | - |
? | |
ginsenoside F2 + H2O | 91% activity compared to ginsenoside Rb1 | Thermotoga petrophila | ginsenoside 20(S)-Rh2 + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | - |
Thermotoga petrophila | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | best ginsenoside substrate | Thermotoga petrophila | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | - |
Thermotoga petrophila DSM 13995 | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | best ginsenoside substrate | Thermotoga petrophila DSM 13995 | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | - |
Thermotoga petrophila RKU-1 | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | best ginsenoside substrate | Thermotoga petrophila RKU-1 | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | - |
Thermotoga petrophila ATCC BAA-488 | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | best ginsenoside substrate | Thermotoga petrophila ATCC BAA-488 | ginsenoside Rd + D-glucopyranose | - |
? | |
ginsenoside Rb1 + H2O | direct conversion of Rb1 into Rg3 | Thermotoga petrophila | ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | - |
Thermotoga petrophila | ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | - |
Thermotoga petrophila DSM 13995 | ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | - |
Thermotoga petrophila RKU-1 | ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | - |
Thermotoga petrophila ATCC BAA-488 | ginsenoside Rg3 + D-glucopyranose | - |
? | |
ginsenoside Rd + H2O | 84% activity compared to ginsenoside Rb1 | Thermotoga petrophila | ginsenoside 20(S)-Rg3 + D-glucopyranose | - |
? | |
ginsenoside Re + H2O | 81% activity compared to ginsenoside Rb1 | Thermotoga petrophila | ginsenoside Rg2 + D-glucopyranose | - |
? | |
ginsenoside Rg1 + H2O | 64% activity compared to ginsenoside Rb1 | Thermotoga petrophila | ginsenoside 20(S)-Rh1 + D-glucopyranose | - |
? | |
additional information | the beta-glucosidase shows preference for cleaving the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1, which produces the pharmacologically active minor ginsenoside 20(S)-Rg3, via graded hydrolysis of the two glucoses at C-20 or direct hydrolysis of the inner glucoses at C-20. The ginsenoside Rg3 exists as two optical isomers, ginsenoside 20(S)-Rg3 and 20(R)-Rg3. The 20(S)-Rg3 isomer is superior to the 20(R)-Rg3 isomer in terms of its water solubility and bioavailability because of the spatial arrangement of the hydroxyl group on carbon 20. No or poor activity with ginsenosides Rb2, Rc, (S)-Rh2, (S)-Rg3, Rg2, and (S)-Rh1 as substrates, no activity with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-alpha--L-arabinofuranoside, 4-nitrophenyl-alpha-L-arabinopyranoside, 4-nitrophenyl-alpha-L-rhamnopyranoside, 4-nitrophenyl-beta-D-xylopyranoside. Low activity with gentiobiose and cellobiose. No activity with laminaribiose and sucrose. The enzyme also has broadly specific beta-glucosidase activity against a wide range of substrates with different glycosidic bonds, including aryl-beta-glucosidic bonds and alkyl-beta-glucosidic bonds, and oligosaccharides | Thermotoga petrophila | ? | - |
? | |
additional information | the beta-glucosidase shows preference for cleaving the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1, which produces the pharmacologically active minor ginsenoside 20(S)-Rg3, via graded hydrolysis of the two glucoses at C-20 or direct hydrolysis of the inner glucoses at C-20. The ginsenoside Rg3 exists as two optical isomers, ginsenoside 20(S)-Rg3 and 20(R)-Rg3. The 20(S)-Rg3 isomer is superior to the 20(R)-Rg3 isomer in terms of its water solubility and bioavailability because of the spatial arrangement of the hydroxyl group on carbon 20. No or poor activity with ginsenosides Rb2, Rc, (S)-Rh2, (S)-Rg3, Rg2, and (S)-Rh1 as substrates, no activity with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-alpha--L-arabinofuranoside, 4-nitrophenyl-alpha-L-arabinopyranoside, 4-nitrophenyl-alpha-L-rhamnopyranoside, 4-nitrophenyl-beta-D-xylopyranoside. Low activity with gentiobiose and cellobiose. No activity with laminaribiose and sucrose. The enzyme also has broadly specific beta-glucosidase activity against a wide range of substrates with different glycosidic bonds, including aryl-beta-glucosidic bonds and alkyl-beta-glucosidic bonds, and oligosaccharides | Thermotoga petrophila DSM 13995 | ? | - |
? | |
additional information | the beta-glucosidase shows preference for cleaving the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1, which produces the pharmacologically active minor ginsenoside 20(S)-Rg3, via graded hydrolysis of the two glucoses at C-20 or direct hydrolysis of the inner glucoses at C-20. The ginsenoside Rg3 exists as two optical isomers, ginsenoside 20(S)-Rg3 and 20(R)-Rg3. The 20(S)-Rg3 isomer is superior to the 20(R)-Rg3 isomer in terms of its water solubility and bioavailability because of the spatial arrangement of the hydroxyl group on carbon 20. No or poor activity with ginsenosides Rb2, Rc, (S)-Rh2, (S)-Rg3, Rg2, and (S)-Rh1 as substrates, no activity with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-alpha--L-arabinofuranoside, 4-nitrophenyl-alpha-L-arabinopyranoside, 4-nitrophenyl-alpha-L-rhamnopyranoside, 4-nitrophenyl-beta-D-xylopyranoside. Low activity with gentiobiose and cellobiose. No activity with laminaribiose and sucrose. The enzyme also has broadly specific beta-glucosidase activity against a wide range of substrates with different glycosidic bonds, including aryl-beta-glucosidic bonds and alkyl-beta-glucosidic bonds, and oligosaccharides | Thermotoga petrophila RKU-1 | ? | - |
? | |
additional information | the beta-glucosidase shows preference for cleaving the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1, which produces the pharmacologically active minor ginsenoside 20(S)-Rg3, via graded hydrolysis of the two glucoses at C-20 or direct hydrolysis of the inner glucoses at C-20. The ginsenoside Rg3 exists as two optical isomers, ginsenoside 20(S)-Rg3 and 20(R)-Rg3. The 20(S)-Rg3 isomer is superior to the 20(R)-Rg3 isomer in terms of its water solubility and bioavailability because of the spatial arrangement of the hydroxyl group on carbon 20. No or poor activity with ginsenosides Rb2, Rc, (S)-Rh2, (S)-Rg3, Rg2, and (S)-Rh1 as substrates, no activity with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-alpha--L-arabinofuranoside, 4-nitrophenyl-alpha-L-arabinopyranoside, 4-nitrophenyl-alpha-L-rhamnopyranoside, 4-nitrophenyl-beta-D-xylopyranoside. Low activity with gentiobiose and cellobiose. No activity with laminaribiose and sucrose. The enzyme also has broadly specific beta-glucosidase activity against a wide range of substrates with different glycosidic bonds, including aryl-beta-glucosidic bonds and alkyl-beta-glucosidic bonds, and oligosaccharides | Thermotoga petrophila ATCC BAA-488 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 81000, recombinant His-tagged enzyme, SDS-PAGE, x * 81243, sequence calculation | Thermotoga petrophila |
Synonyms | Comment | Organism |
---|---|---|
More | cf. EC 3.2.1.21 | Thermotoga petrophila |
Tpebgl3 | - |
Thermotoga petrophila |
Tpet_0898 | - |
Thermotoga petrophila |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
90 | - |
recombinant enzyme | Thermotoga petrophila |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | 98 | over 50% of maximal activity within this range | Thermotoga petrophila |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
purified recombinant His-tagged enzyme, pH 5.0, about 90% activity remaining after 180 min | Thermotoga petrophila |
80 | - |
purified recombinant His-tagged enzyme, pH 5.0, about 70% activity remaining after 180 min | Thermotoga petrophila |
90 | - |
purified recombinant His-tagged enzyme, pH 5.0, about 45% activity remaining after 180 min | Thermotoga petrophila |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5 | - |
recombinant enzyme | Thermotoga petrophila |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
4 | 6 | over 50% of maximal activity within this range | Thermotoga petrophila |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
3.5 | 7.5 | purified recombinant His-tagged enzyme, completely stable at pH 7.5, 50% activity at pH 3.5 | Thermotoga petrophila |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Thermotoga petrophila | sequence calculation | - |
5.28 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the glycosyl hydrolase family 3, GH3. Alignment of the Tpebgl3 cluster with several GH3 beta-glucosidases indicates that they share some conserved motifs: CIHKFV (residues 162-167), GFVMSDWYAGDN (residues 238-249) and IVISRISGEGYDRK (residues 452-463) | Thermotoga petrophila |