Activating Compound | Comment | Organism | Structure |
---|---|---|---|
ethanol | 5% (v/v), 158.4% activity compared to activity without addition of ethanol, 1.25 mM 4-nitrophenyl-beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
methanol | 5% (v/v), 141% activity compared to activity without addition of methanol, 1.25 mM 4-nitrophenyl-beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli and purified | uncultured rumen bacterium |
Crystallization (Comment) | Organism |
---|---|
molecular docking studies for the enzyme with the substrates cellobiose and 4-nitrophenyl beta-D-glucopyranoside. The essential catalytic residues of RuBG3B are D272 and E481. Besides D88, K193, H194, and R160 also form hydrogen bonds with substrates and are conservative. The active center for beta-glucosidic and beta-xylosidic substrates are the same and the bifunctional activity should come from one functional domain. RuBG3A and RuBG3B exhibit higher scores in the docking analysis against 4-nitrophenyl-beta-D-glucopyranoside than against 4-nitrophenyl-beta-D-xylopyranoside, indicating a larger affinity between the enzymes and 4-nitrophenyl-beta-D-glucopyranoside, which is consistent with the kinetic parameters | uncultured rumen bacterium |
molecular docking studies for the enzyme with the substrates cellobiose and 4-nitrophenyl-beta-D-glucopyranoside. The essential catalytic residues of RuBG3A are D297 and E499. Besides D113, K218, H219, and R185 also form hydrogen bonds with substrates and are condervative. The active center for beta-glucosidic and beta-xylosidic substrates are the same and the bifunctional activity should come from one functional domain. RuBG3A and RuBG3B exhibit higher scores in the docking analysis against 4-nitrophenyl-beta-D-glucopyranoside than against 4-nitrophenyl-beta-D-xylopyranoside, indicating a larger affinity between the enzymes and 4-nitrophenyl-beta-D-glucopyranoside, which is consistent with the kinetic parameters | uncultured rumen bacterium |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.22 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 4.6, 42°C | uncultured rumen bacterium | |
1.06 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 4.6, 40°C | uncultured rumen bacterium |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
CaCl2 | 5 mM, 107.3% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
CaCl2 | 5 mM, 92.8% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
CoCl2 | 5 mM, 79.2% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
CoCl2 | 5 mM, 85.5% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
CuCl2 | 5 mM, 79.5% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
CuCl2 | 5 mM, 83.6% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
D-fructose | 5 mM, 102.3% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
D-glucose | 5 mM,82% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
D-xylose | 5 mM, 106.3% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
fructose | 5 mM, 107.6% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
glucose | 5 mM,24.5% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
MgCl2 | 5 mM, 110.1% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
MgCl2 | 5 mM, 96.5% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
MnCl2 | 5 mM, 102.4% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
MnCl2 | 5 mM, 112% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
NiCl2 | 5 mM, 76.7% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
NiCl2 | 5 mM, 86.5% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium | |
xylose | 5 mM, 88% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
ZnSO4 | 5 mM, 111.4% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium | |
ZnSO4 | 5 mM, 90.2% activity compared to activity without addition of MgCl2, 1.25 mM 4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
Organic Solvent | Comment | Organism |
---|---|---|
Ethanol | 5% (v/v), 100.8% activity compared to activity without addition of ethanol, 1.25 mM 4-nitrophenyl-beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium |
Ethanol | 5% (v/v), 158.4% activity compared to activity without addition of ethanol, 1.25 mM 4-nitrophenyl-beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
Methanol | 5% (v/v), 108.2% activity compared to activity without addition of methanol, 1.25 mM 4-nitrophenyl-beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium |
Methanol | 5% (v/v), 141% activity compared to activity without addition of methanol, 1.25 mM 4-nitrophenyl-beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
uncultured rumen bacterium | G3BJX3 | Rubg3A, together with Rubg3B, beta-glucosidase/xylosidase activity, EC 3.2.1.37 and 3.2.1.74; isolated from rumen from a yak rumen metagenomic library, chinese yak, Bos grunniens | - |
uncultured rumen bacterium | G3BJX4 | Rubg3B, together with Rubg3A, beta-glucosidase/xylosidase activity, EC 3.2.1.37 and 3.2.1.74; isolated from rumen from a yak rumen metagenomic library, chinese yak, Bos grunniens | - |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | uncultured rumen bacterium |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | isolated from rumen from a yak rumen metagenomic library, chinese yak, Bos grunniens | uncultured rumen bacterium | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.35 | - |
2-nitrophenyl beta-D-galactopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium |
0.65 | - |
2-nitrophenyl beta-D-galactopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
5.3 | - |
cellobiose as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
11.5 | - |
4-methylumbelliferyl beta-D-cellobioside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
26 | - |
4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 40°C | uncultured rumen bacterium |
33.5 | - |
cellobiose as substrate, pH 4.6, 40°C | uncultured rumen bacterium |
41 | - |
4-methylumbelliferyl beta-D-cellobioside as substrate, pH 4.6, 40°C | uncultured rumen bacterium |
77 | - |
4-nitrophenyl beta-D-glucopyranoside as substrate, pH 4.6, 42°C | uncultured rumen bacterium |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-nitrophenyl beta-D-galactopyranoside + H2O | 0.8% relative activity compared to 4-nitrophenyl beta-D-glucopyranoside as substrate | uncultured rumen bacterium | 2-nitrophenol + D-galactose | - |
? | |
2-nitrophenyl beta-D-galactopyranoside + H2O | 1.3% relative activity compared to 4-nitrophenyl beta-D-glucopyranoside as substrate | uncultured rumen bacterium | 2-nitrophenol + D-galactose | - |
? | |
4-methylumbelliferyl beta-D-cellobioside + 2 H2O | i.e.MuC, 14.9% relative activity compared to 4-nitrophenyl-beta-D-glucopyranoside as substrate | uncultured rumen bacterium | 4-methylumbelliferol + 2 D-glucose | - |
? | |
4-methylumbelliferyl-beta-D-cellobioside + H2O | i.e.MuC, 157.7% relative activity compared to 4-nitrophenyl beta-D-glucopyranoside as substrate | uncultured rumen bacterium | D-glucose + ? | - |
? | |
4-nitrophenyl beta-D-glucopyranoside + H2O | - |
uncultured rumen bacterium | 4-nitrophenol + D-glucose | - |
? | |
cellobiose + H2O | 128.8% relative activity compared to 4-nitrophenyl beta-D-glucopyranoside as substrate | uncultured rumen bacterium | 2 D-glucose | cellobiose is completely hydrolyzed to glucose, evaluated by chromatographic analysis | ? | |
cellobiose + H2O | 6.9% relative activity compared to 4-nitrophenyl-beta-D-glucopyranoside as substrate | uncultured rumen bacterium | 2 D-glucose | 31.5% cellobiose is hydrolyzed to glucose, evaluated by chromatographic analysis | ? | |
cellotetraose + H2O | - |
uncultured rumen bacterium | glucose + cellobiose + cellotriose | with a conversion ratio of 80% | ? | |
additional information | no activity with 4-nitrophenyl alpha-D-glucopyranoside and 4-methylumbelliferyl alpha-D-galactopyranoside as substrates | uncultured rumen bacterium | ? | - |
? | |
additional information | no activity with 4-nitrophenyl alpha-D-glucopyranoside, cellotetraose, and 4-methylumbelliferyl alpha-D-galactopyranoside as substrates | uncultured rumen bacterium | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
beta-glucosidase | - |
uncultured rumen bacterium |
beta-glucosidase/xylosidase | bifunctional, belongs to to the glycoside hydrolase 3 familiy, GH3 | uncultured rumen bacterium |
Rubg3A | gene name, together with Rubg3B, beta-glucosidase/xylosidase activity, EC 3.2.1.37 and 3.2.1.74 | uncultured rumen bacterium |
Rubg3B | gene name, together with Rubg3A, beta-glucosidase/xylosidase activity, EC 3.2.1.37 and 3.2.1.74 | uncultured rumen bacterium |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at, optimal for 4-nitrophenyl beta-D-glucopyranoside | uncultured rumen bacterium |
42 | - |
assay at, optimal for 4-nitrophenyl beta-D-glucopyranoside as substrate | uncultured rumen bacterium |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
43 | - |
Tm-value | uncultured rumen bacterium |
45 | - |
40% activity retained after incubation for 1 h | uncultured rumen bacterium |
45 | - |
stable after 1 h incubation | uncultured rumen bacterium |
50 | - |
Tm-value, retained 50% activity after 20 min incubation | uncultured rumen bacterium |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
126.6 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 4.6, 40°C | uncultured rumen bacterium | |
203 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 4.6, 42°C | uncultured rumen bacterium |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.6 | - |
assay at, optimal for 4-nitrophenol beta-D-glucopyranoside as substrate | uncultured rumen bacterium |
4.6 | - |
assay at, optimal for 4-nitrophenyl beta-D-glucopyranoside as substrate | uncultured rumen bacterium |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11900 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 4.6, 40°C | uncultured rumen bacterium | |
915700 | - |
4-nitrophenyl beta-D-glucopyranoside | pH 4.6, 42°C | uncultured rumen bacterium |