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Literature summary for 3.2.1.74 extracted from

  • Bao, L.; Huang, Q.; Chang, L.; Sun, Q.; Zhou, J.; Lu, H.
    Cloning and characterization of two beta-glucosidase/xylosidase enzymes from yak rumen metagenome (2012), Appl. Biochem. Biotechnol., 166, 72-86.
    View publication on PubMed

Activating Compound

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(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli and purified uncultured rumen bacterium

Crystallization (Commentary)

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 [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

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 Stability

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

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 (Commentary)

Purification (Comment) Organism
recombinant enzyme uncultured rumen bacterium

Source Tissue

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 [micromol/min/mg]

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 and Products (Substrate)

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

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 [°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 [°C]

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 [1/s]

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

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 [mM/s]

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