Literature summary for 3.2.1.156 extracted from

Valenzuela, S.V.; Lopez, S.; Biely, P.; Sanz-Aparicio, J.; Pastor, F.I.
The glycoside hydrolase family 8 reducing-end xylose-releasing exo-oligoxylanase Rex8A from Paenibacillus barcinonensis BP-23 is active on branched xylooligosaccharides (2016), Appl. Environ. Microbiol., 82, 5116-5124 .

Search for more literature for 3.2.1.156

Cloned(Commentary)

Cloned (Comment)	Organism
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

Protein Variants

Protein Variants	Comment	Organism
E70A	site-directed mutagenesis	Paenibacillus barcinonensis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
1.64	-	xylotriose	pH 7.0, 40°C, recombinant wild-type enzyme	Paenibacillus barcinonensis

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
additional information	the enzyme contains no signal peptide	Paenibacillus barcinonensis	-	-

Organism

Organism	UniProt	Comment	Textmining
Paenibacillus barcinonensis	A0A0S2UQQ5	-	-
Paenibacillus barcinonensis BP-23	A0A0S2UQQ5	-	-
Paenibacillus barcinonensis CECT 7022	A0A0S2UQQ5	-	-
Paenibacillus barcinonensis DSM 15478	A0A0S2UQQ5	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 Star (DE3) by nickel affinity chromatography to homogeneity	Paenibacillus barcinonensis

Reaction

Reaction	Comment	Organism	Reaction ID
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	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-nitrophenyl-beta-D-xylopyranoside + H2O	-	Paenibacillus barcinonensis	4-nitrophenol + D-xylopyranose	-	?
4-nitrophenyl-beta-D-xylopyranoside + H2O	-	Paenibacillus barcinonensis CECT 7022	4-nitrophenol + D-xylopyranose	-	?
4-nitrophenyl-beta-D-xylopyranoside + H2O	-	Paenibacillus barcinonensis BP-23	4-nitrophenol + D-xylopyranose	-	?
4-nitrophenyl-beta-D-xylopyranoside + H2O	-	Paenibacillus barcinonensis DSM 15478	4-nitrophenol + D-xylopyranose	-	?
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	?	-	?
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	?	-	?
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	?	-	?
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	?	-	?
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	?	-	?
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	?	-	?
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	?	-	?
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	?	-	?
xylan + H2O	very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan	Paenibacillus barcinonensis	?	-	?
xylan + H2O	very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan	Paenibacillus barcinonensis CECT 7022	?	-	?
xylan + H2O	very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan	Paenibacillus barcinonensis BP-23	?	-	?
xylan + H2O	very low activity, beechwood xylan, oat spelt xylan, or birchwood xylan	Paenibacillus barcinonensis DSM 15478	?	-	?
xylohexaose + H2O	-	Paenibacillus barcinonensis	4 D-xylose + xylobiose	-	?
xylopentaose + H2O	-	Paenibacillus barcinonensis	3 D-xylose + xylobiose	-	?
xylotetraose + H2O	-	Paenibacillus barcinonensis	2 D-xylose + xylobiose	-	?
xylotriose + H2O	preferred substrate	Paenibacillus barcinonensis	D-xylose + xylobiose	-	?
xylotriose + H2O	preferred substrate	Paenibacillus barcinonensis CECT 7022	D-xylose + xylobiose	-	?
xylotriose + H2O	preferred substrate	Paenibacillus barcinonensis BP-23	D-xylose + xylobiose	-	?
xylotriose + H2O	preferred substrate	Paenibacillus barcinonensis DSM 15478	D-xylose + xylobiose	-	?

Subunits

Subunits	Comment	Organism
?	x * 44217, sequence calculation	Paenibacillus barcinonensis

Synonyms

Synonyms	Comment	Organism
DFQ00_11062	-	Paenibacillus barcinonensis
reducing-end xylose-releasing exo-oligoxylanase	-	Paenibacillus barcinonensis
Rex8A	-	Paenibacillus barcinonensis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
40	-	recombinant enzyme	Paenibacillus barcinonensis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
118.8	-	xylotriose	pH 7.0, 40°C, recombinant wild-type enzyme	Paenibacillus barcinonensis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	recombinant enzyme	Paenibacillus barcinonensis

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	10	activity range, profile overview	Paenibacillus barcinonensis

pI Value

Organism	Comment	pI Value Maximum	pI Value
Paenibacillus barcinonensis	sequence calculation	-	5.25

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the glycoside hydrolase family 8, GH8	Paenibacillus barcinonensis
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
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