Cloned (Comment) | Organism |
---|---|
gene PgXynB, subcloning in Escherichia coli strain DH5alpha, expression of GST- or His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Penicillium griseofulvum |
Protein Variants | Comment | Organism |
---|---|---|
K131S/K132S | site-directed mutagenesis, with or without deletion mutation DELTAP130, the mutant without deletion shows no sensitivity to inhibitor XIP-1, like the wild-type enzyme, but shows increased activity compared to the wild-type, the mutant with deletion mutation is sensitive to inhibitor XIP-1 | Penicillium griseofulvum |
additional information | generation of XynB mutans variants, e.g. deletion mutant DELTAP130, displaying increased catalytic efficiency towards wheat arabinoxylan and xylo-oligosaccharides and identified specific determinants in PgXynB thumb region responsible for resistance to the wheat xylanase inhibitor XIP-I | Penicillium griseofulvum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
XIP-I | a wheat xylanase inhibitor, that inhibits only XynB mutant DELTAP130/K131S/K132S, but not mutant K131S/K132S and the wild-type XynB | Penicillium griseofulvum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics, kinetic analysis and homology modeling, overview | Penicillium griseofulvum | |
0.103 | - |
xylotetraose | pH 4.5, 30°C, wild-type XynB | Penicillium griseofulvum | |
0.183 | - |
xylotetraose | pH 4.5, 30°C, XynB mutant DELTAP130/K131S/K132S | Penicillium griseofulvum | |
0.25 | - |
xylotetraose | pH 4.5, 30°C, XynB mutant K131S/K132S | Penicillium griseofulvum | |
1.23 | - |
xylopentaose | pH 4.5, 30°C, wild-type XynB | Penicillium griseofulvum | |
3 | - |
xylopentaose | pH 4.5, 30°C, XynB mutant DELTAP130/K131S/K132S | Penicillium griseofulvum | |
3.17 | - |
xylopentaose | pH 4.5, 30°C, XynB mutant K131S/K132S | Penicillium griseofulvum | |
13.33 | - |
xylohexaose | pH 4.5, 30°C, wild-type XynB | Penicillium griseofulvum | |
20 | - |
xylohexaose | pH 4.5, 30°C, XynB mutant DELTAP130/K131S/K132S | Penicillium griseofulvum | |
25 | - |
xylohexaose | pH 4.5, 30°C, XynB mutant K131S/K132S | Penicillium griseofulvum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
21300 | - |
x * 21300, recombinant XynB, SDS-PAGE and sequence calculation | Penicillium griseofulvum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Penicillium griseofulvum | - |
gene PgXynB | - |
Purification (Comment) | Organism |
---|---|
recombinant GST- or His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3), the His-tagged enzymes by nickel affinity chromatography, the GST-tagged by glutathione affinity chromatography and cleavage of the tag by thrombin. The specific activity of thrombin-cleaved PgXynB is 5fold higher than His6-tagged PgXynB | Penicillium griseofulvum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
arabinoxylan + H2O | wheat arabinoxylan substrates with different arabinose to xylose ratio | Penicillium griseofulvum | D-xylose + ? | - |
? | |
additional information | catalytic residue is E85. No activity with xylotriose by XynB. Molecular determinants of substrate and inhibitor specificities of the GH11 enzyme, kinetic analysis and homology modeling, overview | Penicillium griseofulvum | ? | - |
? | |
xylohexaose + H2O | - |
Penicillium griseofulvum | D-xylose + ? | - |
? | |
xylopentaose + H2O | - |
Penicillium griseofulvum | 5 D-xylose + ? | - |
? | |
xylotetraose + H2O | - |
Penicillium griseofulvum | D-xylose + ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 21300, recombinant XynB, SDS-PAGE and sequence calculation | Penicillium griseofulvum |
More | XynB structure homology molecular modelling, overview | Penicillium griseofulvum |
Synonyms | Comment | Organism |
---|---|---|
More | the enzyme belongs to the glycosyl hydrolase family 11, GH11 | Penicillium griseofulvum |
xylanase | - |
Penicillium griseofulvum |
XynB | - |
Penicillium griseofulvum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Penicillium griseofulvum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 50 | - |
Penicillium griseofulvum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
purified recombinant PgXynB is stable at 30 °C, pH 4.5 for 240 min, with more than 85% of the activity remaining | Penicillium griseofulvum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1104 | - |
arabinoxylan | pH 4.5, 30°C, mutant DELTAP130 | Penicillium griseofulvum | |
1166 | - |
arabinoxylan | pH 4.5, 30°C, wild-type XynB | Penicillium griseofulvum | |
1537 | - |
arabinoxylan | pH 4.5, 30°C, mutant DELTAP130/K131S/K132S | Penicillium griseofulvum | |
1592 | - |
arabinoxylan | pH 4.5, 30°C, mutant K131S/K132S | Penicillium griseofulvum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
- |
Penicillium griseofulvum |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
2.6 | 7.6 | 50% of maximal activity at pH 3.5 to pH 5.5 | Penicillium griseofulvum |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4.5 | - |
purified recombinant PgXynB is stable at 30 °C, pH 4.5 for 240 min, with more than 85% of the activity remaining | Penicillium griseofulvum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0165 | - |
XIP-I | pH 4.5, 30°C, mutant DELTAP130/K131S/K132S | Penicillium griseofulvum |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Penicillium griseofulvum | recombinant His6-tagged wild-type enzyme, isoelectric focusing | - |
6.8 |