BRENDA - Enzyme Database

Biochemical analysis of a beta-D-xylosidase and a bifunctional xylanase-ferulic acid esterase from a xylanolytic gene cluster in Prevotella ruminicola 23

Dodd, D.; Kocherginskaya, S.A.; Spies, M.A.; Beery, K.E.; Abbas, C.A.; Mackie, R.I.; Cann, I.K.; J. Bacteriol. 191, 3328-3338 (2009) View publication on PubMedView publication on EuropePMC

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
3.1.1.73
Xyn10D-Fae1A amplicons cloned into the pGEM-T vector via TA cloning and subcloned in frame with the hexahistidine-encoding sequence of a modified pET-28a expression vector by replacing the NdeI-XhoI polylinker, expressed in Escherichia coli BL-21 CodonPlus (DE3) RIL competent cells
Prevotella ruminicola
3.2.1.37
ecxpression in Escherichia coli
Prevotella ruminicola
Engineering
EC Number
Protein Variants
Commentary
Organism
3.1.1.73
E280S
the ferulic acid esterase activity increases 3fold. Capacity for Xyn10D-Fae1A to depolymerize oat spelt xylan is attenuated
Prevotella ruminicola
3.1.1.73
S629A
the ferulic acid esterase activity for Xyn10D-Fae1A is attenuated. Depolymerizes xylan to an extent similar to the that of wild-type
Prevotella ruminicola
3.2.1.37
D269A
no catalytic activity
Prevotella ruminicola
3.2.1.37
D269N
no catalytic activity
Prevotella ruminicola
3.2.1.37
E594Q
activity similar to wild-type
Prevotella ruminicola
3.2.1.37
E616A
about 5% of wild-type activity
Prevotella ruminicola
3.2.1.37
R149L
no catalytic activity
Prevotella ruminicola
3.2.1.37
Y237F
less than 10% of wild-type activity
Prevotella ruminicola
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.2.1.37
1.8
-
4-nitrophenyl-beta-D-xylopyranoside
wild-type, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
2
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E594Q, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
3.9
-
4-nitrophenyl-beta-D-xylopyranoside
mutant Y237F, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
18
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E616A, pH 5.0, 37°C
Prevotella ruminicola
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.1.73
84000
-
1 * 84000, hexahistidine fusion protein, SDS-PAGE, exists either as monomer or homodimer in solution
Prevotella ruminicola
3.1.1.73
84000
-
2 * 84000, gel filtration, exists either as monomer or homodimer in solution
Prevotella ruminicola
3.1.1.73
124000
-
gel filtration
Prevotella ruminicola
Organism
EC Number
Organism
UniProt
Commentary
Textmining
3.1.1.73
Prevotella ruminicola
-
-
-
3.1.1.73
Prevotella ruminicola 23
-
-
-
3.2.1.37
Prevotella ruminicola
D5EY15
isoform Xyl3A
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
3.1.1.73
Xyn10D-Fae1A purified by metal affinity chromatography and gel filtration
Prevotella ruminicola
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
3.1.1.73
1-naphthyl butyrate + H2O
esterase activity
708923
Prevotella ruminicola
butyric acid + 1-naphthol
-
-
-
?
3.1.1.73
1-naphthyl butyrate + H2O
esterase activity
708923
Prevotella ruminicola 23
butyric acid + 1-naphthol
-
-
-
?
3.1.1.73
feruloyl xylooligosaccharide + H2O
degrees of polymerization (X3 to X6) are hydrolyzed mostly to xylobiose, with some xylose also being released
708923
Prevotella ruminicola
ferulic acid + xylooligosaccharide
-
-
-
?
3.1.1.73
feruloyl xylooligosaccharide + H2O
degrees of polymerization (X3 to X6) are hydrolyzed mostly to xylobiose, with some xylose also being released
708923
Prevotella ruminicola 23
ferulic acid + xylooligosaccharide
-
-
-
?
3.1.1.73
feruloylated oat spelt xylan + H2O
xylanase activity
708923
Prevotella ruminicola
ferulic acid + ?
-
-
-
?
3.1.1.73
feruloylated oat spelt xylan + H2O
xylanase activity
708923
Prevotella ruminicola 23
ferulic acid + ?
-
-
-
?
3.1.1.73
methyl ferulate + H2O
-
708923
Prevotella ruminicola
ferulic acid + methanol
-
-
-
?
3.1.1.73
methyl ferulate + H2O
-
708923
Prevotella ruminicola 23
ferulic acid + methanol
-
-
-
?
3.1.1.73
additional information
no xylobiose hydrolysis to xylose
708923
Prevotella ruminicola
?
-
-
-
?
3.1.1.73
additional information
no xylobiose hydrolysis to xylose
708923
Prevotella ruminicola 23
?
-
-
-
?
3.2.1.37
4-nitrophenyl alpha-L-arabinofuranoside + H2O
poor substrate
708923
Prevotella ruminicola
4-nitrophenol + alpha-L-arabinofuranose
-
-
-
?
3.2.1.37
4-nitrophenyl beta-D-xylopyranoside + H2O
-
708923
Prevotella ruminicola
4-nitrophenol + beta-D-xylose
-
-
-
?
3.2.1.37
additional information
no substrate: 4-nitrophenly-beta-D-glucopyranoside. When incubated in combination with bifunctional xylanase-ferulic acid esterase Xyn10D-Fae1A, Xyl3A improves the release of xylose monomers from a hemicellulosic xylan substrate, suggesting that these two enzymes function synergistically to depolymerize xylan
708923
Prevotella ruminicola
?
-
-
-
?
3.2.1.37
xylobiose + H2O
-
708923
Prevotella ruminicola
2 D-xylose
-
-
-
?
3.2.1.37
xylohexaose + H2O
sole products released are xylose monomers
708923
Prevotella ruminicola
6 D-xylose
-
-
-
?
3.2.1.37
xylopentaose + H2O
sole products released are xylose monomers
708923
Prevotella ruminicola
5 D-xylose
-
-
-
?
3.2.1.37
xylotetraose + H2O
-
708923
Prevotella ruminicola
4 D-xylose
-
-
-
?
3.2.1.37
xylotriose + H2O
-
708923
Prevotella ruminicola
3 D-xylose
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.1.1.73
homodimer
2 * 84000, gel filtration, exists either as monomer or homodimer in solution
Prevotella ruminicola
3.1.1.73
monomer
1 * 84000, hexahistidine fusion protein, SDS-PAGE, exists either as monomer or homodimer in solution
Prevotella ruminicola
Synonyms
EC Number
Synonyms
Commentary
Organism
3.1.1.73
xylanase-ferulic acid esterase
-
Prevotella ruminicola
3.1.1.73
Xyn10D-Fae1A
-
Prevotella ruminicola
3.2.1.37
Xyl3A
-
Prevotella ruminicola
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.2.1.37
1.6
-
4-nitrophenyl-beta-D-xylopyranoside
mutant Y237F, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
3.3
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E616A, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
9.7
-
4-nitrophenyl-beta-D-xylopyranoside
wild-type, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
10
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E594Q, pH 5.0, 37°C
Prevotella ruminicola
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.1.73
5
-
-
Prevotella ruminicola
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.73
Xyn10D-Fae1A amplicons cloned into the pGEM-T vector via TA cloning and subcloned in frame with the hexahistidine-encoding sequence of a modified pET-28a expression vector by replacing the NdeI-XhoI polylinker, expressed in Escherichia coli BL-21 CodonPlus (DE3) RIL competent cells
Prevotella ruminicola
3.2.1.37
ecxpression in Escherichia coli
Prevotella ruminicola
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
3.1.1.73
E280S
the ferulic acid esterase activity increases 3fold. Capacity for Xyn10D-Fae1A to depolymerize oat spelt xylan is attenuated
Prevotella ruminicola
3.1.1.73
S629A
the ferulic acid esterase activity for Xyn10D-Fae1A is attenuated. Depolymerizes xylan to an extent similar to the that of wild-type
Prevotella ruminicola
3.2.1.37
D269A
no catalytic activity
Prevotella ruminicola
3.2.1.37
D269N
no catalytic activity
Prevotella ruminicola
3.2.1.37
E594Q
activity similar to wild-type
Prevotella ruminicola
3.2.1.37
E616A
about 5% of wild-type activity
Prevotella ruminicola
3.2.1.37
R149L
no catalytic activity
Prevotella ruminicola
3.2.1.37
Y237F
less than 10% of wild-type activity
Prevotella ruminicola
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.2.1.37
1.8
-
4-nitrophenyl-beta-D-xylopyranoside
wild-type, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
2
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E594Q, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
3.9
-
4-nitrophenyl-beta-D-xylopyranoside
mutant Y237F, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
18
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E616A, pH 5.0, 37°C
Prevotella ruminicola
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.1.1.73
84000
-
1 * 84000, hexahistidine fusion protein, SDS-PAGE, exists either as monomer or homodimer in solution
Prevotella ruminicola
3.1.1.73
84000
-
2 * 84000, gel filtration, exists either as monomer or homodimer in solution
Prevotella ruminicola
3.1.1.73
124000
-
gel filtration
Prevotella ruminicola
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.1.1.73
Xyn10D-Fae1A purified by metal affinity chromatography and gel filtration
Prevotella ruminicola
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
3.1.1.73
1-naphthyl butyrate + H2O
esterase activity
708923
Prevotella ruminicola
butyric acid + 1-naphthol
-
-
-
?
3.1.1.73
1-naphthyl butyrate + H2O
esterase activity
708923
Prevotella ruminicola 23
butyric acid + 1-naphthol
-
-
-
?
3.1.1.73
feruloyl xylooligosaccharide + H2O
degrees of polymerization (X3 to X6) are hydrolyzed mostly to xylobiose, with some xylose also being released
708923
Prevotella ruminicola
ferulic acid + xylooligosaccharide
-
-
-
?
3.1.1.73
feruloyl xylooligosaccharide + H2O
degrees of polymerization (X3 to X6) are hydrolyzed mostly to xylobiose, with some xylose also being released
708923
Prevotella ruminicola 23
ferulic acid + xylooligosaccharide
-
-
-
?
3.1.1.73
feruloylated oat spelt xylan + H2O
xylanase activity
708923
Prevotella ruminicola
ferulic acid + ?
-
-
-
?
3.1.1.73
feruloylated oat spelt xylan + H2O
xylanase activity
708923
Prevotella ruminicola 23
ferulic acid + ?
-
-
-
?
3.1.1.73
methyl ferulate + H2O
-
708923
Prevotella ruminicola
ferulic acid + methanol
-
-
-
?
3.1.1.73
methyl ferulate + H2O
-
708923
Prevotella ruminicola 23
ferulic acid + methanol
-
-
-
?
3.1.1.73
additional information
no xylobiose hydrolysis to xylose
708923
Prevotella ruminicola
?
-
-
-
?
3.1.1.73
additional information
no xylobiose hydrolysis to xylose
708923
Prevotella ruminicola 23
?
-
-
-
?
3.2.1.37
4-nitrophenyl alpha-L-arabinofuranoside + H2O
poor substrate
708923
Prevotella ruminicola
4-nitrophenol + alpha-L-arabinofuranose
-
-
-
?
3.2.1.37
4-nitrophenyl beta-D-xylopyranoside + H2O
-
708923
Prevotella ruminicola
4-nitrophenol + beta-D-xylose
-
-
-
?
3.2.1.37
additional information
no substrate: 4-nitrophenly-beta-D-glucopyranoside. When incubated in combination with bifunctional xylanase-ferulic acid esterase Xyn10D-Fae1A, Xyl3A improves the release of xylose monomers from a hemicellulosic xylan substrate, suggesting that these two enzymes function synergistically to depolymerize xylan
708923
Prevotella ruminicola
?
-
-
-
?
3.2.1.37
xylobiose + H2O
-
708923
Prevotella ruminicola
2 D-xylose
-
-
-
?
3.2.1.37
xylohexaose + H2O
sole products released are xylose monomers
708923
Prevotella ruminicola
6 D-xylose
-
-
-
?
3.2.1.37
xylopentaose + H2O
sole products released are xylose monomers
708923
Prevotella ruminicola
5 D-xylose
-
-
-
?
3.2.1.37
xylotetraose + H2O
-
708923
Prevotella ruminicola
4 D-xylose
-
-
-
?
3.2.1.37
xylotriose + H2O
-
708923
Prevotella ruminicola
3 D-xylose
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.1.1.73
homodimer
2 * 84000, gel filtration, exists either as monomer or homodimer in solution
Prevotella ruminicola
3.1.1.73
monomer
1 * 84000, hexahistidine fusion protein, SDS-PAGE, exists either as monomer or homodimer in solution
Prevotella ruminicola
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.2.1.37
1.6
-
4-nitrophenyl-beta-D-xylopyranoside
mutant Y237F, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
3.3
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E616A, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
9.7
-
4-nitrophenyl-beta-D-xylopyranoside
wild-type, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
10
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E594Q, pH 5.0, 37°C
Prevotella ruminicola
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.1.1.73
5
-
-
Prevotella ruminicola
General Information
EC Number
General Information
Commentary
Organism
3.1.1.73
physiological function
Xyn10D-Fae1A possesses both endo-beta-1,4-xylanase and ferulic acid esterase activities, Glu280 is an important residue for xylanase activity and Ser629 is an important residue for ferulic acid esterase activity. When incubated in combination with Xyn10D-Fae1A, the beta-D-glucosidase Xyl3A improves the release of xylose monomers from a hemicellulosic xylan substrate, thus these two enzymes function synergistically to depolymerize xylan. Two catalytic domains for Xyn10D-Fae1A are functionally coupled
Prevotella ruminicola
3.2.1.37
physiological function
when incubated in combination with bifunctional xylanase-ferulic acid esterase Xyn10D-Fae1A, Xyl3A improves the release of xylose monomers from a hemicellulosic xylan substrate, suggesting that these two enzymes function synergistically to depolymerize xylan
Prevotella ruminicola
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.1.1.73
physiological function
Xyn10D-Fae1A possesses both endo-beta-1,4-xylanase and ferulic acid esterase activities, Glu280 is an important residue for xylanase activity and Ser629 is an important residue for ferulic acid esterase activity. When incubated in combination with Xyn10D-Fae1A, the beta-D-glucosidase Xyl3A improves the release of xylose monomers from a hemicellulosic xylan substrate, thus these two enzymes function synergistically to depolymerize xylan. Two catalytic domains for Xyn10D-Fae1A are functionally coupled
Prevotella ruminicola
3.2.1.37
physiological function
when incubated in combination with bifunctional xylanase-ferulic acid esterase Xyn10D-Fae1A, Xyl3A improves the release of xylose monomers from a hemicellulosic xylan substrate, suggesting that these two enzymes function synergistically to depolymerize xylan
Prevotella ruminicola
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.2.1.37
0.2
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E616A, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
0.41
-
4-nitrophenyl-beta-D-xylopyranoside
mutant Y237F, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
5
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E594Q, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
5.4
-
4-nitrophenyl-beta-D-xylopyranoside
wild-type, pH 5.0, 37°C
Prevotella ruminicola
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
3.2.1.37
0.2
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E616A, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
0.41
-
4-nitrophenyl-beta-D-xylopyranoside
mutant Y237F, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
5
-
4-nitrophenyl-beta-D-xylopyranoside
mutant E594Q, pH 5.0, 37°C
Prevotella ruminicola
3.2.1.37
5.4
-
4-nitrophenyl-beta-D-xylopyranoside
wild-type, pH 5.0, 37°C
Prevotella ruminicola