Literature summary for 3.2.1.45 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 .

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

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

Inhibitors

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

KM Value [mM]

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

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Thermoanaerobacterium xylanolyticum	F6BL85	-	-
Thermoanaerobacterium xylanolyticum GH116	F6BL85	-	-

Purification (Commentary)

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

Source Tissue

Source Tissue	Comment	Organism	Textmining

Specific Activity [micromol/min/mg]

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

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-nitrophenyl beta-cellobioside + H2O	1.1% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	4-nitrophenol + cellobiose	-	?
4-nitrophenyl beta-D-galactoside + H2O	33% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	4-nitrophenol + beta-D-galactose	-	?
4-nitrophenyl beta-D-galactoside + H2O	33% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum GH116	4-nitrophenol + beta-D-galactose	-	?
4-nitrophenyl beta-D-glucoside + H2O	100% activity	Thermoanaerobacterium xylanolyticum	4-nitrophenol + beta-D-glucose	-	?
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	-	?
cellobiose + H2O	49% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
cellohexaose + H2O	49% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
cellotetraose + H2O	55% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
cellotriose + H2O	57% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
cellotriose + H2O	57% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum GH116	?	-	?
gentibiose + H2O	8.2% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
glucosylceramide + H2O	-	Thermoanaerobacterium xylanolyticum	beta-D-glucose + ceramide	-	?
glucosylceramide + H2O	-	Thermoanaerobacterium xylanolyticum GH116	beta-D-glucose + ceramide	-	?
glucosyllysosphingosine + H2O	-	Thermoanaerobacterium xylanolyticum	?	-	?
laminaribiose + H2O	43% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	2 D-glucose	-	?
laminaripentaose + H2O	60% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
laminaritetraose + H2O	55% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
laminaritriose + H2O	59% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	?	-	?
laminaritriose + H2O	59% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum GH116	?	-	?
methyl beta-D-glucoside + H2O	0.3% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	methanol + D-glucose	-	?
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	?	-	?
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	?	-	?
n-heptyl beta-D-glucoside + H2O	14% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	heptanol + D-glucose	-	?
n-octyl beta-D-glucoside + H2O	13% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	octanol + D-glucose	-	?
sophorose + H2O	15% activity compared to 4-nitrophenyl beta-D-glucoside	Thermoanaerobacterium xylanolyticum	2 D-glucose	-	?

Synonyms

Synonyms	Comment	Organism
beta-glucosidase	-	Thermoanaerobacterium xylanolyticum

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5.5	-	-	Thermoanaerobacterium xylanolyticum

kcat/KM [mM/s]

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