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Literature summary extracted from

  • Pei, J.; Xie, J.; Yin, R.; Zhao, L.; Ding, G.; Wang, Z.; Xiao, W.
    Enzymatic transformation of ginsenoside Rb1 to ginsenoside 20(S)-Rg3 by GH3 beta-glucosidase from Thermotoga thermarum DSM 5069 T (2015), J. Mol. Catal. B, 113, 104-109 .
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
3.2.1.192 Al3+ 20% activation at 1 mM Pseudothermotoga thermarum
3.2.1.192 Mn2+ 41% activation at 1 mM Pseudothermotoga thermarum
3.2.1.192 additional information no effect by 10 mM EDTA Pseudothermotoga thermarum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.192 gene Theth_1706, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of His-tagged enzyme in Escherichia coli strain JM109(DE3) Pseudothermotoga thermarum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.192 Ag+ 81% inhibition at 1 mM Pseudothermotoga thermarum
3.2.1.192 Cu2+ 20% inhibition at 1 mM Pseudothermotoga thermarum
3.2.1.192 ethanol inhibits 88% at 20% v/v, 30% at 10% v/v Pseudothermotoga thermarum
3.2.1.192 Fe2+ 23% inhibition at 1 mM Pseudothermotoga thermarum
3.2.1.192 Hg2+ 94% inhibition at 1 mM Pseudothermotoga thermarum
3.2.1.192 methanol inhibits 94% at 20% v/v, 12% at 10% v/v Pseudothermotoga thermarum
3.2.1.192 additional information no effect by 10 mM EDTA Pseudothermotoga thermarum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.192 0.065
-
4-nitrophenyl beta-D-glucopyranoside pH 5.0, 90°C, recombinant enzyme Pseudothermotoga thermarum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.192 additional information no or poor effects by 1 mM of Ni2+, Fe3+, Mg2+, Zn2+, K+, Ca2+, Li+, and Co2+ Pseudothermotoga thermarum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.192 ginsenoside Rb1 + H2O Pseudothermotoga thermarum
-
ginsenoside Rd + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O Pseudothermotoga thermarum direct conversion of Rb1 into Rg3 ginsenoside Rg3 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O Pseudothermotoga thermarum DSM 5069 T
-
ginsenoside Rd + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rd + H2O Pseudothermotoga thermarum
-
ginsenoside Rg3 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rd + H2O Pseudothermotoga thermarum DSM 5069 T
-
ginsenoside Rg3 + D-glucopyranose
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.192 Pseudothermotoga thermarum F7YVT1
-
-
3.2.1.192 Pseudothermotoga thermarum DSM 5069 T F7YVT1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.192 recombinant His-tagged enzyme 2.9fold from Escherichia coli strain BL21(DE3) by heat treatment and nickel affinity chromatography Pseudothermotoga thermarum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.192 additional information
-
at a concentration of 0.75 U/mL of the enzyme, a temperature of 85°C and pH 5.0,10 g/l ginsenoside Rb1 is transformed into 6.9 g/l Rg3 within 60 min with a corresponding molar conversion of 97.8% Pseudothermotoga thermarum
3.2.1.192 70.7
-
purified recombinant His-tagged enzyme, pH 5.0, 90°C, substrate is 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.192 4-nitrophenyl beta-D-glucopyranoside + H2O
-
Pseudothermotoga thermarum 4-nitrophenol + beta-D-glucopyranose
-
?
3.2.1.192 4-nitrophenyl beta-D-glucopyranoside + H2O
-
Pseudothermotoga thermarum DSM 5069 T 4-nitrophenol + beta-D-glucopyranose
-
?
3.2.1.192 ginsenoside compound K + H2O 33% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum 20(S)-protopanaxadiol + D-glucopyranose
-
?
3.2.1.192 ginsenoside F2 + H2O 34% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum ginsenoside 20(S)-Rh2 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O
-
Pseudothermotoga thermarum ginsenoside Rd + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O 118% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum ginsenoside Rd + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O
-
Pseudothermotoga thermarum DSM 5069 T ginsenoside Rd + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O 118% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum DSM 5069 T ginsenoside Rd + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rb1 + H2O direct conversion of Rb1 into Rg3 Pseudothermotoga thermarum ginsenoside Rg3 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rd + H2O
-
Pseudothermotoga thermarum ginsenoside Rg3 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rd + H2O
-
Pseudothermotoga thermarum DSM 5069 T ginsenoside Rg3 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rd + H2O 43% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum ginsenoside 20(S)-Rg3 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Re + H2O 110% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum ginsenoside Rg2 + D-glucopyranose
-
?
3.2.1.192 ginsenoside Rg1 + H2O 81% activity compared to 4-nitrophenyl beta-D-glucopyranoside Pseudothermotoga thermarum ginsenoside 20(S)-Rh1 + D-glucopyranose
-
?
3.2.1.192 additional information the beta-glucosidase exhibits high selectivity to cleave the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1 and catalyzes the conversion of ginsenoside Rb1 or Rd to the more pharmacologically active minor ginsenoside 20(S)-Rg3. The enzyme is also able to hydrolyze cellobiose and gentiobiose. No activity is detected with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-beta-D-xylopyranoside, 4-nitrophenyl-alpha-L-arabinofuranoside, carboxymethyl cellulose, and sucrose Pseudothermotoga thermarum ?
-
?
3.2.1.192 additional information the beta-glucosidase exhibits high selectivity to cleave the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1 and catalyzes the conversion of ginsenoside Rb1 or Rd to the more pharmacologically active minor ginsenoside 20(S)-Rg3. The enzyme is also able to hydrolyze cellobiose and gentiobiose. No activity is detected with 4-nitrophenyl-beta-D-galactopyranoside, 4-nitrophenyl-beta-D-xylopyranoside, 4-nitrophenyl-alpha-L-arabinofuranoside, carboxymethyl cellulose, and sucrose Pseudothermotoga thermarum DSM 5069 T ?
-
?

Subunits

EC Number Subunits Comment Organism
3.2.1.192 ? x * 82000, recombinant His-tagged enzyme, SDS-PAGE Pseudothermotoga thermarum

Synonyms

EC Number Synonyms Comment Organism
3.2.1.192 BGL3T
-
Pseudothermotoga thermarum
3.2.1.192 GH3 beta-glucosidase
-
Pseudothermotoga thermarum
3.2.1.192 More cf. EC 3.2.1.21 Pseudothermotoga thermarum
3.2.1.192 Theth_1706
-
Pseudothermotoga thermarum
3.2.1.192 Tt-BGL
-
Pseudothermotoga thermarum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.192 90
-
recombinant enzyme Pseudothermotoga thermarum

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.2.1.192 65 100 activity profile of the recombinant enzyme, overview Pseudothermotoga thermarum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.192 85
-
purified recombinant His-tagged enzyme, pH 5.0, about 90% activity remaining after 120 min Pseudothermotoga thermarum
3.2.1.192 90
-
purified recombinant His-tagged enzyme, pH 5.0, 50% activity remaining after 60 min, 40% after 120 min Pseudothermotoga thermarum
3.2.1.192 95
-
purified recombinant His-tagged enzyme, pH 5.0, about 10% activity remaining after 120 min, 20% after 60 min Pseudothermotoga thermarum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.192 121
-
4-nitrophenyl beta-D-glucopyranoside pH 5.0, 90°C, recombinant enzyme Pseudothermotoga thermarum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.192 5
-
recombinant enzyme Pseudothermotoga thermarum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.2.1.192 4.5 6.5 over 50% of maximal activity within this range Pseudothermotoga thermarum

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.2.1.192 4.5 7.5 purified recombinant His-tagged enzyme, 80% activity remaining at pH 4.5-7.5 Pseudothermotoga thermarum
3.2.1.192 5.5
-
purified recombinant His-tagged enzyme, completely stable at Pseudothermotoga thermarum

General Information

EC Number General Information Comment Organism
3.2.1.192 evolution the enzyme belongs to the glycosyl hydrolase family 3, GH3 Pseudothermotoga thermarum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.192 1861
-
4-nitrophenyl beta-D-glucopyranoside pH 5.0, 90°C, recombinant enzyme Pseudothermotoga thermarum