Literature summary 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 .

Activating Compound

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

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
3.2.1.192	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

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

Inhibitors

EC Number	Inhibitors	Comment	Organism
3.2.1.192	Al3+	67% inhibition at 5 mM	Thermotoga petrophila
3.2.1.192	Cu2+	40% inhibition at 5 mM	Thermotoga petrophila
3.2.1.192	DMSO	inhibits 26% at 15% and 42% at 30%	Thermotoga petrophila
3.2.1.192	ethanol	inhibits 20% at 15% and 45% at 30%	Thermotoga petrophila
3.2.1.192	Hg2+	complete inhibition at 1-5 mM	Thermotoga petrophila
3.2.1.192	methanol	activates 14% at 5%, inhibits 20% at 30%	Thermotoga petrophila
3.2.1.192	additional information	no significant effect of 5 mM EDTA on the enzyme activity	Thermotoga petrophila
3.2.1.192	NH4+	45% inhibition at 5 mM	Thermotoga petrophila

Metals/Ions

EC Number	Metals/Ions	Comment	Organism
3.2.1.192	Ca2+	activates about 1.5fold at 5 mM, stabilizes the purified recombinant enzyme	Thermotoga petrophila
3.2.1.192	Co2+	activates about 1.5fold at 5 mM, does not stabilize the purified recombinant enzyme	Thermotoga petrophila
3.2.1.192	Mn2+	activates about 1.5fold at 5 mM, stabilizes the purified recombinant enzyme	Thermotoga petrophila

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
3.2.1.192	ginsenoside Rb1 + H2O	Thermotoga petrophila	-	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	Thermotoga petrophila	direct conversion of Rb1 into Rg3	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	Thermotoga petrophila DSM 13995	-	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	Thermotoga petrophila RKU-1	-	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	Thermotoga petrophila ATCC BAA-488	-	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	Thermotoga petrophila	-	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	Thermotoga petrophila DSM 13995	-	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	Thermotoga petrophila RKU-1	-	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	Thermotoga petrophila ATCC BAA-488	-	ginsenoside Rg3 + D-glucopyranose	-	?

Organism

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

Purification (Commentary)

EC Number	Purification (Comment)	Organism
3.2.1.192	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]

EC Number	Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
3.2.1.192	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
3.2.1.192	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)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3.2.1.192	2-nitrophenyl beta-D-glucopyranoside + H2O	low activity	Thermotoga petrophila	2-nitrophenol + beta-D-glucopyranose	-	?
3.2.1.192	2-nitrophenyl beta-D-glucopyranoside + H2O	low activity	Thermotoga petrophila DSM 13995	2-nitrophenol + beta-D-glucopyranose	-	?
3.2.1.192	2-nitrophenyl beta-D-glucopyranoside + H2O	low activity	Thermotoga petrophila RKU-1	2-nitrophenol + beta-D-glucopyranose	-	?
3.2.1.192	2-nitrophenyl beta-D-glucopyranoside + H2O	low activity	Thermotoga petrophila ATCC BAA-488	2-nitrophenol + beta-D-glucopyranose	-	?
3.2.1.192	4-nitrophenyl beta-D-glucopyranoside + H2O	-	Thermotoga petrophila	4-nitrophenol + beta-D-glucopyranose	-	?
3.2.1.192	ginsenoside compound K + H2O	35% activity compared to ginsenoside Rb1	Thermotoga petrophila	20(S)-protopanaxadiol + D-glucopyranose	-	?
3.2.1.192	ginsenoside F2 + H2O	91% activity compared to ginsenoside Rb1	Thermotoga petrophila	ginsenoside 20(S)-Rh2 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	-	Thermotoga petrophila	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	best ginsenoside substrate	Thermotoga petrophila	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	-	Thermotoga petrophila DSM 13995	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	best ginsenoside substrate	Thermotoga petrophila DSM 13995	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	-	Thermotoga petrophila RKU-1	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	best ginsenoside substrate	Thermotoga petrophila RKU-1	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	-	Thermotoga petrophila ATCC BAA-488	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	best ginsenoside substrate	Thermotoga petrophila ATCC BAA-488	ginsenoside Rd + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rb1 + H2O	direct conversion of Rb1 into Rg3	Thermotoga petrophila	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	-	Thermotoga petrophila	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	-	Thermotoga petrophila DSM 13995	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	-	Thermotoga petrophila RKU-1	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	-	Thermotoga petrophila ATCC BAA-488	ginsenoside Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rd + H2O	84% activity compared to ginsenoside Rb1	Thermotoga petrophila	ginsenoside 20(S)-Rg3 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Re + H2O	81% activity compared to ginsenoside Rb1	Thermotoga petrophila	ginsenoside Rg2 + D-glucopyranose	-	?
3.2.1.192	ginsenoside Rg1 + H2O	64% activity compared to ginsenoside Rb1	Thermotoga petrophila	ginsenoside 20(S)-Rh1 + D-glucopyranose	-	?
3.2.1.192	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	?	-	?
3.2.1.192	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	?	-	?
3.2.1.192	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	?	-	?
3.2.1.192	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

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

Synonyms

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

Temperature Optimum [°C]

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

Temperature Range [°C]

EC Number	Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
3.2.1.192	75	98	over 50% of maximal activity within this range	Thermotoga petrophila

Temperature Stability [°C]

EC Number	Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
3.2.1.192	70	-	purified recombinant His-tagged enzyme, pH 5.0, about 90% activity remaining after 180 min	Thermotoga petrophila
3.2.1.192	80	-	purified recombinant His-tagged enzyme, pH 5.0, about 70% activity remaining after 180 min	Thermotoga petrophila
3.2.1.192	90	-	purified recombinant His-tagged enzyme, pH 5.0, about 45% activity remaining after 180 min	Thermotoga petrophila

pH Optimum

EC Number	pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
3.2.1.192	5	-	recombinant enzyme	Thermotoga petrophila

pH Range

EC Number	pH Minimum	pH Maximum	Comment	Organism
3.2.1.192	4	6	over 50% of maximal activity within this range	Thermotoga petrophila

pH Stability

EC Number	pH Stability	pH Stability Maximum	Comment	Organism
3.2.1.192	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

EC Number	Organism	Comment	pI Value Maximum	pI Value
3.2.1.192	Thermotoga petrophila	sequence calculation	-	5.28

General Information

EC Number	General Information	Comment	Organism
3.2.1.192	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