Cloning and characterization of arabinoxylan arabinofuranofydrolases-D3 (AXHd3) from Bifidobacterium adolescentis DSM20083

L.A.M. van den Broek, R.M. Lloyd, G. Beldman, J.C. Verdoes, B.V. McCleary, A.G.J. Voragen

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Abstract

Arabinoxylan arabinofuranohydrolase-D3 (AXHd3) from Bifidobacterium adolescentis releases only C3-linked arabinose residues from double-substituted xylose residues. A genomic library of B. adolescentis DSM20083 was screened for the presence of the axhD3 gene. Two plasmids were identified containing part of the axhD3 gene. The nucleotide sequences were combined and three open reading frames (ORFs) were found. The first ORF showed high homology with xylanases belonging to family 8 of the glycoside hydrolases and this gene was designated xylA. The second ORF was the axhD3 gene belonging to glycoside hydrolase family 43. The third (partial) ORF coded for a putative carboxylesterase. The axhD3 gene was cloned and expressed in Escherichia coli. Several substrates were employed in the biochemical characterization of recombinant AXHd3. The enzyme showed the highest activity toward wheat arabinoxylan oligosaccharides. In addition, -xylanase from Trichoderma sp. was able to degrade soluble wheat arabinoxylan polymer to a higher extent, after pretreatment with recombinant AXHd3. Arabinoxylan oligosaccharides incubated with a combination of recombinant AXHd3 and an -l-arabinofuranosidase from Aspergillus niger did not result in a higher maximal release of arabinose than incubation with these enzymes separately.
Original languageEnglish
Pages (from-to)641-647
JournalApplied Microbiology and Biotechnology
Volume67
Issue number5
DOIs
Publication statusPublished - 2005

Fingerprint

Organism Cloning
Open Reading Frames
Arabinose
Genes
Glycoside Hydrolases
Oligosaccharides
Triticum
Carboxylesterase
Trichoderma
Genomic Library
Aspergillus niger
Xylose
Enzymes
Polymers
Plasmids
Bifidobacterium adolescentis
arabinoxylan
alpha-N-arabinofuranosidase
Escherichia coli

Keywords

  • intestinal anaerobic bacterium
  • alpha-l-arabinofuranosidase
  • breve k-110
  • oligosaccharides
  • purification
  • longum
  • fermentation
  • degradation
  • xylosidase
  • hydrolases

Cite this

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title = "Cloning and characterization of arabinoxylan arabinofuranofydrolases-D3 (AXHd3) from Bifidobacterium adolescentis DSM20083",
abstract = "Arabinoxylan arabinofuranohydrolase-D3 (AXHd3) from Bifidobacterium adolescentis releases only C3-linked arabinose residues from double-substituted xylose residues. A genomic library of B. adolescentis DSM20083 was screened for the presence of the axhD3 gene. Two plasmids were identified containing part of the axhD3 gene. The nucleotide sequences were combined and three open reading frames (ORFs) were found. The first ORF showed high homology with xylanases belonging to family 8 of the glycoside hydrolases and this gene was designated xylA. The second ORF was the axhD3 gene belonging to glycoside hydrolase family 43. The third (partial) ORF coded for a putative carboxylesterase. The axhD3 gene was cloned and expressed in Escherichia coli. Several substrates were employed in the biochemical characterization of recombinant AXHd3. The enzyme showed the highest activity toward wheat arabinoxylan oligosaccharides. In addition, -xylanase from Trichoderma sp. was able to degrade soluble wheat arabinoxylan polymer to a higher extent, after pretreatment with recombinant AXHd3. Arabinoxylan oligosaccharides incubated with a combination of recombinant AXHd3 and an -l-arabinofuranosidase from Aspergillus niger did not result in a higher maximal release of arabinose than incubation with these enzymes separately.",
keywords = "intestinal anaerobic bacterium, alpha-l-arabinofuranosidase, breve k-110, oligosaccharides, purification, longum, fermentation, degradation, xylosidase, hydrolases",
author = "{van den Broek}, L.A.M. and R.M. Lloyd and G. Beldman and J.C. Verdoes and B.V. McCleary and A.G.J. Voragen",
year = "2005",
doi = "10.1007/s00253-004-1850-9",
language = "English",
volume = "67",
pages = "641--647",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Verlag",
number = "5",

}

Cloning and characterization of arabinoxylan arabinofuranofydrolases-D3 (AXHd3) from Bifidobacterium adolescentis DSM20083. / van den Broek, L.A.M.; Lloyd, R.M.; Beldman, G.; Verdoes, J.C.; McCleary, B.V.; Voragen, A.G.J.

In: Applied Microbiology and Biotechnology, Vol. 67, No. 5, 2005, p. 641-647.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Cloning and characterization of arabinoxylan arabinofuranofydrolases-D3 (AXHd3) from Bifidobacterium adolescentis DSM20083

AU - van den Broek, L.A.M.

AU - Lloyd, R.M.

AU - Beldman, G.

AU - Verdoes, J.C.

AU - McCleary, B.V.

AU - Voragen, A.G.J.

PY - 2005

Y1 - 2005

N2 - Arabinoxylan arabinofuranohydrolase-D3 (AXHd3) from Bifidobacterium adolescentis releases only C3-linked arabinose residues from double-substituted xylose residues. A genomic library of B. adolescentis DSM20083 was screened for the presence of the axhD3 gene. Two plasmids were identified containing part of the axhD3 gene. The nucleotide sequences were combined and three open reading frames (ORFs) were found. The first ORF showed high homology with xylanases belonging to family 8 of the glycoside hydrolases and this gene was designated xylA. The second ORF was the axhD3 gene belonging to glycoside hydrolase family 43. The third (partial) ORF coded for a putative carboxylesterase. The axhD3 gene was cloned and expressed in Escherichia coli. Several substrates were employed in the biochemical characterization of recombinant AXHd3. The enzyme showed the highest activity toward wheat arabinoxylan oligosaccharides. In addition, -xylanase from Trichoderma sp. was able to degrade soluble wheat arabinoxylan polymer to a higher extent, after pretreatment with recombinant AXHd3. Arabinoxylan oligosaccharides incubated with a combination of recombinant AXHd3 and an -l-arabinofuranosidase from Aspergillus niger did not result in a higher maximal release of arabinose than incubation with these enzymes separately.

AB - Arabinoxylan arabinofuranohydrolase-D3 (AXHd3) from Bifidobacterium adolescentis releases only C3-linked arabinose residues from double-substituted xylose residues. A genomic library of B. adolescentis DSM20083 was screened for the presence of the axhD3 gene. Two plasmids were identified containing part of the axhD3 gene. The nucleotide sequences were combined and three open reading frames (ORFs) were found. The first ORF showed high homology with xylanases belonging to family 8 of the glycoside hydrolases and this gene was designated xylA. The second ORF was the axhD3 gene belonging to glycoside hydrolase family 43. The third (partial) ORF coded for a putative carboxylesterase. The axhD3 gene was cloned and expressed in Escherichia coli. Several substrates were employed in the biochemical characterization of recombinant AXHd3. The enzyme showed the highest activity toward wheat arabinoxylan oligosaccharides. In addition, -xylanase from Trichoderma sp. was able to degrade soluble wheat arabinoxylan polymer to a higher extent, after pretreatment with recombinant AXHd3. Arabinoxylan oligosaccharides incubated with a combination of recombinant AXHd3 and an -l-arabinofuranosidase from Aspergillus niger did not result in a higher maximal release of arabinose than incubation with these enzymes separately.

KW - intestinal anaerobic bacterium

KW - alpha-l-arabinofuranosidase

KW - breve k-110

KW - oligosaccharides

KW - purification

KW - longum

KW - fermentation

KW - degradation

KW - xylosidase

KW - hydrolases

U2 - 10.1007/s00253-004-1850-9

DO - 10.1007/s00253-004-1850-9

M3 - Article

VL - 67

SP - 641

EP - 647

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 5

ER -