EC Number | Cloned (Comment) | Organism |
---|---|---|
3.2.1.156 | gene rex8A, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 Star (DE3) | Paenibacillus barcinonensis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.2.1.156 | E70A | site-directed mutagenesis | Paenibacillus barcinonensis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.156 | 1.64 | - |
xylotriose | pH 7.0, 40°C, recombinant wild-type enzyme | Paenibacillus barcinonensis |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
3.2.1.156 | additional information | the enzyme contains no signal peptide | Paenibacillus barcinonensis | - |
- |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.2.1.156 | Paenibacillus barcinonensis | A0A0S2UQQ5 | - |
- |
3.2.1.156 | Paenibacillus barcinonensis BP-23 | A0A0S2UQQ5 | - |
- |
3.2.1.156 | Paenibacillus barcinonensis CECT 7022 | A0A0S2UQQ5 | - |
- |
3.2.1.156 | Paenibacillus barcinonensis DSM 15478 | A0A0S2UQQ5 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.2.1.156 | recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 Star (DE3) by nickel affinity chromatography to homogeneity | Paenibacillus barcinonensis |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
3.2.1.156 | beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranosyl-(1-4)-beta-D-xylopyranose + H2O = 2 beta-D-xylopyranosyl-(1-4)-alpha-D-xylopyranose | processivemodeof hydrolysis, overview. The xylooligosaccharides are progressively degraded from Xn to Xn-1 and then subsequently degraded in the same way to xylose and xylobiose | Paenibacillus barcinonensis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.2.1.156 | 4-nitrophenyl-beta-D-xylopyranoside + H2O | - |
Paenibacillus barcinonensis | 4-nitrophenol + D-xylopyranose | - |
? | |
3.2.1.156 | 4-nitrophenyl-beta-D-xylopyranoside + H2O | - |
Paenibacillus barcinonensis CECT 7022 | 4-nitrophenol + D-xylopyranose | - |
? | |
3.2.1.156 | 4-nitrophenyl-beta-D-xylopyranoside + H2O | - |
Paenibacillus barcinonensis BP-23 | 4-nitrophenol + D-xylopyranose | - |
? | |
3.2.1.156 | 4-nitrophenyl-beta-D-xylopyranoside + H2O | - |
Paenibacillus barcinonensis DSM 15478 | 4-nitrophenol + D-xylopyranose | - |
? | |
3.2.1.156 | 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose + H2O | docking analysis of Rex8A with methyl-glucuronic acid branched oligomers. Mixtures containing the aldouronic acids 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl3) (aldotetraouronic acid) and beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-[4-O-methyl-alpha-D-glucuronosyl-(1->2)]-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl5) (aldohexaouronic acid) are prepared from beechwood 4-O-methyl-D-glucuronoxylan by xylanase treatment. Aldotetraouronic acid MeGlcA3Xyl3 consists of a xylotriose with a methyl-glucuronic acid substituent in the third xylose from the reducing end. The cleavage of this substrate to xylose and to an aldouronic acid shortened by one residue indicates that Rex8A accommodates the methyl-glucuronic acid-substituted xylopyranosyl residue in the -2 subsite of the catalytic cleft of the enzyme. Ligand accommodation in the catalytic site, a xylotriose decorated with a 4-O-MeGlcA moiety at O2 of the third xylose from the reducing end (MeGlcA3Xyl3) is modeled into the active-site channel | Paenibacillus barcinonensis | ? | - |
? | |
3.2.1.156 | 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose + H2O | docking analysis of Rex8A with methyl-glucuronic acid branched oligomers. Mixtures containing the aldouronic acids 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl3) (aldotetraouronic acid) and beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-[4-O-methyl-alpha-D-glucuronosyl-(1->2)]-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl5) (aldohexaouronic acid) are prepared from beechwood 4-O-methyl-D-glucuronoxylan by xylanase treatment. Aldotetraouronic acid MeGlcA3Xyl3 consists of a xylotriose with a methyl-glucuronic acid substituent in the third xylose from the reducing end. The cleavage of this substrate to xylose and to an aldouronic acid shortened by one residue indicates that Rex8A accommodates the methyl-glucuronic acid-substituted xylopyranosyl residue in the -2 subsite of the catalytic cleft of the enzyme. Ligand accommodation in the catalytic site, a xylotriose decorated with a 4-O-MeGlcA moiety at O2 of the third xylose from the reducing end (MeGlcA3Xyl3) is modeled into the active-site channel | Paenibacillus barcinonensis CECT 7022 | ? | - |
? | |
3.2.1.156 | 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose + H2O | docking analysis of Rex8A with methyl-glucuronic acid branched oligomers. Mixtures containing the aldouronic acids 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl3) (aldotetraouronic acid) and beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-[4-O-methyl-alpha-D-glucuronosyl-(1->2)]-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl5) (aldohexaouronic acid) are prepared from beechwood 4-O-methyl-D-glucuronoxylan by xylanase treatment. Aldotetraouronic acid MeGlcA3Xyl3 consists of a xylotriose with a methyl-glucuronic acid substituent in the third xylose from the reducing end. The cleavage of this substrate to xylose and to an aldouronic acid shortened by one residue indicates that Rex8A accommodates the methyl-glucuronic acid-substituted xylopyranosyl residue in the -2 subsite of the catalytic cleft of the enzyme. Ligand accommodation in the catalytic site, a xylotriose decorated with a 4-O-MeGlcA moiety at O2 of the third xylose from the reducing end (MeGlcA3Xyl3) is modeled into the active-site channel | Paenibacillus barcinonensis BP-23 | ? | - |
? | |
3.2.1.156 | 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose + H2O | docking analysis of Rex8A with methyl-glucuronic acid branched oligomers. Mixtures containing the aldouronic acids 4-O-methyl-beta-D-glucuronosyl-(1->2)-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl3) (aldotetraouronic acid) and beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-[4-O-methyl-alpha-D-glucuronosyl-(1->2)]-beta-D-xylopyranosyl-(1->4)-beta-D-xylopyranosyl-(1->4)-D-xylose (MeGlcA3Xyl5) (aldohexaouronic acid) are prepared from beechwood 4-O-methyl-D-glucuronoxylan by xylanase treatment. Aldotetraouronic acid MeGlcA3Xyl3 consists of a xylotriose with a methyl-glucuronic acid substituent in the third xylose from the reducing end. The cleavage of this substrate to xylose and to an aldouronic acid shortened by one residue indicates that Rex8A accommodates the methyl-glucuronic acid-substituted xylopyranosyl residue in the -2 subsite of the catalytic cleft of the enzyme. Ligand accommodation in the catalytic site, a xylotriose decorated with a 4-O-MeGlcA moiety at O2 of the third xylose from the reducing end (MeGlcA3Xyl3) is modeled into the active-site channel | Paenibacillus barcinonensis DSM 15478 | ? | - |
? | |
3.2.1.156 | additional information | the enzyme efficiently hydrolyzes xylooligosaccharides and shows minor activity on polymeric xylan. The enzyme shows also catalytic activity on branched xylooligosaccharides, i.e. the release of xylose from the reducing end. Hydrolysis products from oligosaccharides and xylan are analyzed by thin-layer chromatography (TLC) and MALDI TOF/TOF mass spectrometry. No activity with xylobiose, low activity with xylan | Paenibacillus barcinonensis | ? | - |
? | |
3.2.1.156 | additional information | the enzyme efficiently hydrolyzes xylooligosaccharides and shows minor activity on polymeric xylan. The enzyme shows also catalytic activity on branched xylooligosaccharides, i.e. the release of xylose from the reducing end. Hydrolysis products from oligosaccharides and xylan are analyzed by thin-layer chromatography (TLC) and MALDI TOF/TOF mass spectrometry. No activity with xylobiose, low activity with xylan | Paenibacillus barcinonensis CECT 7022 | ? | - |
? | |
3.2.1.156 | additional information | the enzyme efficiently hydrolyzes xylooligosaccharides and shows minor activity on polymeric xylan. The enzyme shows also catalytic activity on branched xylooligosaccharides, i.e. the release of xylose from the reducing end. Hydrolysis products from oligosaccharides and xylan are analyzed by thin-layer chromatography (TLC) and MALDI TOF/TOF mass spectrometry. No activity with xylobiose, low activity with xylan | Paenibacillus barcinonensis BP-23 | ? | - |
? | |
3.2.1.156 | additional information | the enzyme efficiently hydrolyzes xylooligosaccharides and shows minor activity on polymeric xylan. The enzyme shows also catalytic activity on branched xylooligosaccharides, i.e. the release of xylose from the reducing end. Hydrolysis products from oligosaccharides and xylan are analyzed by thin-layer chromatography (TLC) and MALDI TOF/TOF mass spectrometry. No activity with xylobiose, low activity with xylan | Paenibacillus barcinonensis DSM 15478 | ? | - |
? | |
3.2.1.156 | xylan + H2O | very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan | Paenibacillus barcinonensis | ? | - |
? | |
3.2.1.156 | xylan + H2O | very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan | Paenibacillus barcinonensis CECT 7022 | ? | - |
? | |
3.2.1.156 | xylan + H2O | very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan | Paenibacillus barcinonensis BP-23 | ? | - |
? | |
3.2.1.156 | xylan + H2O | very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan | Paenibacillus barcinonensis DSM 15478 | ? | - |
? | |
3.2.1.156 | xylohexaose + H2O | - |
Paenibacillus barcinonensis | 4 D-xylose + xylobiose | - |
? | |
3.2.1.156 | xylopentaose + H2O | - |
Paenibacillus barcinonensis | 3 D-xylose + xylobiose | - |
? | |
3.2.1.156 | xylotetraose + H2O | - |
Paenibacillus barcinonensis | 2 D-xylose + xylobiose | - |
? | |
3.2.1.156 | xylotriose + H2O | preferred substrate | Paenibacillus barcinonensis | D-xylose + xylobiose | - |
? | |
3.2.1.156 | xylotriose + H2O | preferred substrate | Paenibacillus barcinonensis CECT 7022 | D-xylose + xylobiose | - |
? | |
3.2.1.156 | xylotriose + H2O | preferred substrate | Paenibacillus barcinonensis BP-23 | D-xylose + xylobiose | - |
? | |
3.2.1.156 | xylotriose + H2O | preferred substrate | Paenibacillus barcinonensis DSM 15478 | D-xylose + xylobiose | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.2.1.156 | ? | x * 44217, sequence calculation | Paenibacillus barcinonensis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.2.1.156 | DFQ00_11062 | - |
Paenibacillus barcinonensis |
3.2.1.156 | reducing-end xylose-releasing exo-oligoxylanase | - |
Paenibacillus barcinonensis |
3.2.1.156 | Rex8A | - |
Paenibacillus barcinonensis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.2.1.156 | 40 | - |
recombinant enzyme | Paenibacillus barcinonensis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.2.1.156 | 118.8 | - |
xylotriose | pH 7.0, 40°C, recombinant wild-type enzyme | Paenibacillus barcinonensis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.2.1.156 | 7 | - |
recombinant enzyme | Paenibacillus barcinonensis |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
3.2.1.156 | 4 | 10 | activity range, profile overview | Paenibacillus barcinonensis |
EC Number | Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|---|
3.2.1.156 | Paenibacillus barcinonensis | sequence calculation | - |
5.25 |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.2.1.156 | evolution | the enzyme belongs to the glycoside hydrolase family 8, GH8 | Paenibacillus barcinonensis |
3.2.1.156 | additional information | modeling of the three-dimensional structure of Rex8A shows an (alpha/alpha)6 barrel fold where the loops connecting the alpha-helices contour the active site. These loops, which show high sequence diversity among GH8 enzymes, shape a catalytic cleft with a -2 subsite that can accommodate methyl-glucuronic acid decorations. Putative proton donor is Glu70 and catalytic base is Asp265. Residues Leu320, His321, and Pro322 form the loop structure. Structural molecular modeling of Rex8A | Paenibacillus barcinonensis |
3.2.1.156 | physiological function | the enzyme is involved in depolymerization of glucuronoxylan, a major component of the lignocellulosic substrates. Rex8A is a reducing-end xylose-releasing exo-oligoxylanase that efficiently hydrolyzes xylose from neutral and acidic xylooligosaccharides generated by the action of other xylanases also secreted by the strain. The hydrolytic ability of Rex8A on branched oligomers can be crucial for the complete depolymerization of highly substituted xylans, which is indispensable to accomplish biomass deconstruction and to generate efficient catalysts | Paenibacillus barcinonensis |