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Literature summary for 3.2.1.8 extracted from

  • Cervera Tison, M.; Andre-Leroux, G.; Lafond, M.; Georis, J.; Juge, N.; Berrin, J.G.
    Molecular determinants of substrate and inhibitor specificities of the Penicillium griseofulvum family 11 xylanases (2009), Biochim. Biophys. Acta, 1794, 438-445.
    View publication on PubMed
Search for more literature for 3.2.1.8

Cloned(Commentary)

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

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

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 [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 [Da] Molecular Weight Maximum [Da] Comment Organism
21300
-
 x * 21300, recombinant XynB, SDS-PAGE and sequence calculation Penicillium griseofulvum

Organism

Organism UniProt Comment Textmining
Penicillium griseofulvum
-
 gene PgXynB
-

Purification (Commentary)

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 and Products (Substrate)

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

Subunits Comment Organism
? x * 21300, recombinant XynB, SDS-PAGE and sequence calculation Penicillium griseofulvum
More XynB structure homology molecular modelling, overview Penicillium griseofulvum

Synonyms

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Penicillium griseofulvum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 50
-
 Penicillium griseofulvum

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
4.5
-
-
 Penicillium griseofulvum

pH Range

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 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 [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0165
-
 XIP-I pH 4.5, 30°C, mutant DELTAP130/K131S/K132S Penicillium griseofulvum

pI Value

Organism Comment pI Value Maximum pI Value
Penicillium griseofulvum recombinant His6-tagged wild-type enzyme, isoelectric focusing
-
 6.8