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Literature summary for 3.2.1.192 extracted from

  • Xie, J.; Zhao, D.; Zhao, L.; Pei, J.; Xiao, W.; Ding, G.; Wang, Z.
    Overexpression and characterization of a Ca2+ activated thermostable beta-glucosidase with high ginsenoside Rb1 to ginsenoside 20(S)-Rg3 bioconversion productivity (2015), J. Ind. Microbiol. Biotechnol., 42, 839-850 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
methanol activates 14% at 5%, inhibits 20% at 30% Thermotoga petrophila

Cloned(Commentary)

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

General Stability Organism
Ca2+ and Mn2+ stabilize the purified recombinant enzyme Thermotoga petrophila

Inhibitors

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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Thermotoga petrophila A5IL43
-
-
Thermotoga petrophila ATCC BAA-488 A5IL43
-
-
Thermotoga petrophila DSM 13995 A5IL43
-
-
Thermotoga petrophila RKU-1 A5IL43
-
-

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

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

Subunits Comment Organism
? x * 81000, recombinant His-tagged enzyme, SDS-PAGE, x * 81243, sequence calculation Thermotoga petrophila

Synonyms

Synonyms Comment Organism
More cf. EC 3.2.1.21 Thermotoga petrophila
Tpebgl3
-
Thermotoga petrophila
Tpet_0898
-
Thermotoga petrophila

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
90
-
recombinant enzyme Thermotoga petrophila

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
75 98 over 50% of maximal activity within this range Thermotoga petrophila

Temperature Stability [°C]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
recombinant enzyme Thermotoga petrophila

pH Range

pH Minimum pH Maximum Comment Organism
4 6 over 50% of maximal activity within this range Thermotoga petrophila

pH Stability

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

pI Value

Organism Comment pI Value Maximum pI Value
Thermotoga petrophila sequence calculation
-
5.28

General Information

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