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Literature summary extracted from

  • Charoenwattanasatien, R.; Pengthaisong, S.; Breen, I.; Mutoh, R.; Sansenya, S.; Hua, Y.; Tankrathok, A.; Wu, L.; Songsiriritthigul, C.; Tanaka, H.; Williams, S.J.; Davies, G.J.; Kurisu, G.; Cairns, J.R.
    Bacterial beta-glucosidase reveals the structural and functional basis of genetic defects in human glucocerebrosidase 2 (GBA2) (2016), ACS Chem. Biol., 11, 1891-1900 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.45 expressed in Escherichia coli BL21(DE3) cells Thermoanaerobacterium xylanolyticum

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.2.1.45 hanging drop vapor diffusion method, using 0.2 M ammonium sulfate, 15% (w/v) PEG 6000, 0.1 M Bis-Tris propane, pH 8.0 Thermoanaerobacterium xylanolyticum

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.45 D508H the mutant shows strongly reduced activity compared to the wild type enzyme Thermoanaerobacterium xylanolyticum
3.2.1.45 D508N the mutant shows strongly reduced activity compared to the wild type enzyme Thermoanaerobacterium xylanolyticum
3.2.1.45 D593A the mutant shows strongly reduced activity compared to the wild type enzyme Thermoanaerobacterium xylanolyticum
3.2.1.45 E441A the mutant shows strongly reduced activity compared to the wild type enzyme Thermoanaerobacterium xylanolyticum
3.2.1.45 R544W the mutant shows slightly increased activity compared to the wild type enzyme Thermoanaerobacterium xylanolyticum
3.2.1.45 R786H the mutant shows reduced activity compared to the wild type enzyme Thermoanaerobacterium xylanolyticum

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.45 2,4-dinitrophenyl 2-deoxy-2-fluoro-beta-D-glucoside
-
Thermoanaerobacterium xylanolyticum
3.2.1.45 deoxynojirimycin
-
Thermoanaerobacterium xylanolyticum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.45 0.18
-
4-nitrophenyl beta-D-glucoside wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 0.21
-
4-nitrophenyl beta-D-glucoside wild type enzyme with N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 0.25
-
cellobiose wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 0.26
-
4-nitrophenyl beta-D-glucoside mutant enzyme R544W, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 0.27
-
laminaribiose wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 15.7
-
4-nitrophenyl beta-D-glucoside mutant enzyme R786H, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 16.3
-
4-nitrophenyl beta-D-galactoside wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 41
-
4-nitrophenyl beta-D-glucoside mutant enzyme D508N, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 110
-
4-nitrophenyl beta-D-glucoside mutant enzyme D508H, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.45 glucosylceramide + H2O Thermoanaerobacterium xylanolyticum
-
beta-D-glucose + ceramide
-
?
3.2.1.45 glucosylceramide + H2O Thermoanaerobacterium xylanolyticum GH116
-
beta-D-glucose + ceramide
-
?
3.2.1.45 glucosyllysosphingosine + H2O Thermoanaerobacterium xylanolyticum
-
?
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.45 Thermoanaerobacterium xylanolyticum F6BL85
-
-
3.2.1.45 Thermoanaerobacterium xylanolyticum GH116 F6BL85
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.45 Talon cobalt affinity resin column chromatography, HisTrap column chromatography, and Superdex 200 gel filtration Thermoanaerobacterium xylanolyticum

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.2.1.45 0.0817
-
with methyl beta-D-glucoside as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 0.123
-
with 4-nitrophenyl N-acetyl-beta-D-glucosaminide as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 2.23
-
with gentibiose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 3.58
-
with n-octyl beta-D-glucoside as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 3.74
-
with n-heptyl beta-D-glucoside as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 4.1
-
with sophorose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 8.9
-
with 4-nitrophenyl beta-D-galactoside as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 11.7
-
with laminaribiose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 13.4
-
with cellobiose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 13.5
-
with cellohexaose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 13.8
-
with cellopentaose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 14.9
-
with cellotetraose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 15
-
with laminaritetraose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 15.6
-
with cellotriose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 16.1
-
with laminaritriose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 16.5
-
with laminaripentaose as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 27.3
-
with 4-nitrophenyl beta-D-glucoside as substrate, wild type enzyme, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.45 4-nitrophenyl beta-cellobioside + H2O 1.1% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum 4-nitrophenol + cellobiose
-
?
3.2.1.45 4-nitrophenyl beta-D-galactoside + H2O 33% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum 4-nitrophenol + beta-D-galactose
-
?
3.2.1.45 4-nitrophenyl beta-D-galactoside + H2O 33% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum GH116 4-nitrophenol + beta-D-galactose
-
?
3.2.1.45 4-nitrophenyl beta-D-glucoside + H2O 100% activity Thermoanaerobacterium xylanolyticum 4-nitrophenol + beta-D-glucose
-
?
3.2.1.45 4-nitrophenyl N-acetyl-beta-D-glucosaminide + H2O 0.45% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum 4-nitrophenol + N-acetyl-beta-D-glucosamine
-
?
3.2.1.45 cellobiose + H2O 49% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 cellohexaose + H2O 49% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 cellotetraose + H2O 55% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 cellotriose + H2O 57% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 cellotriose + H2O 57% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum GH116 ?
-
?
3.2.1.45 gentibiose + H2O 8.2% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 glucosylceramide + H2O
-
Thermoanaerobacterium xylanolyticum beta-D-glucose + ceramide
-
?
3.2.1.45 glucosylceramide + H2O
-
Thermoanaerobacterium xylanolyticum GH116 beta-D-glucose + ceramide
-
?
3.2.1.45 glucosyllysosphingosine + H2O
-
Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 laminaribiose + H2O 43% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum 2 D-glucose
-
?
3.2.1.45 laminaripentaose + H2O 60% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 laminaritetraose + H2O 55% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 laminaritriose + H2O 59% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 laminaritriose + H2O 59% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum GH116 ?
-
?
3.2.1.45 methyl beta-D-glucoside + H2O 0.3% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum methanol + D-glucose
-
?
3.2.1.45 additional information the enzyme shows less than 0.04% activity with 4-nitrophenyl alpha-D-galactoside, 4-nitrophenyl alpha-L-arabinoside, 4-nitrophenyl beta-D-fucoside, 4-nitrophenyl alpha-D-glucoside, 4-nitrophenyl beta-D-mannoside, and 4-nitrophenyl beta-D-xyloside Thermoanaerobacterium xylanolyticum ?
-
?
3.2.1.45 additional information the enzyme shows less than 0.04% activity with 4-nitrophenyl alpha-D-galactoside, 4-nitrophenyl alpha-L-arabinoside, 4-nitrophenyl beta-D-fucoside, 4-nitrophenyl alpha-D-glucoside, 4-nitrophenyl beta-D-mannoside, and 4-nitrophenyl beta-D-xyloside Thermoanaerobacterium xylanolyticum GH116 ?
-
?
3.2.1.45 n-heptyl beta-D-glucoside + H2O 14% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum heptanol + D-glucose
-
?
3.2.1.45 n-octyl beta-D-glucoside + H2O 13% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum octanol + D-glucose
-
?
3.2.1.45 sophorose + H2O 15% activity compared to 4-nitrophenyl beta-D-glucoside Thermoanaerobacterium xylanolyticum 2 D-glucose
-
?

Synonyms

EC Number Synonyms Comment Organism
3.2.1.45 beta-glucosidase
-
Thermoanaerobacterium xylanolyticum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.45 75
-
-
Thermoanaerobacterium xylanolyticum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.2.1.45 81.3
-
melting temperature Thermoanaerobacterium xylanolyticum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.45 3.47
-
4-nitrophenyl beta-D-glucoside mutant enzyme D508H, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 30.2
-
4-nitrophenyl beta-D-glucoside mutant enzyme D508N, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 37.6
-
4-nitrophenyl beta-D-glucoside wild type enzyme with N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 41.4
-
laminaribiose wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 44.4
-
cellobiose wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 48.2
-
4-nitrophenyl beta-D-glucoside mutant enzyme R544W, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 49
-
4-nitrophenyl beta-D-glucoside wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 193
-
4-nitrophenyl beta-D-glucoside mutant enzyme R786H, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 255
-
4-nitrophenyl beta-D-galactoside wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.45 5.5
-
-
Thermoanaerobacterium xylanolyticum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.45 0.031
-
4-nitrophenyl beta-D-glucoside mutant enzyme D508H, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 0.74
-
4-nitrophenyl beta-D-glucoside mutant enzyme D508N, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 12.3
-
4-nitrophenyl beta-D-glucoside mutant enzyme R786H, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 15.6
-
4-nitrophenyl beta-D-galactoside wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 153
-
laminaribiose wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 178
-
cellobiose wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 179
-
4-nitrophenyl beta-D-glucoside wild type enzyme with N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 186
-
4-nitrophenyl beta-D-glucoside mutant enzyme R544W, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum
3.2.1.45 272
-
4-nitrophenyl beta-D-glucoside wild type enzyme without N-terminal tag, at pH 5.5 and 60°C Thermoanaerobacterium xylanolyticum