Isolation of cellotriosyl blocks from barley β-glucan with endo-1,4-β-glucanase from Trichoderma reesei

Anu Ajithkumar (Corresponding Author), Roger Andersson, Matti Siika-aho, Maija Tenkanen, Per Åman

Research output: Contribution to journalArticleScientificpeer-review

21 Citations (Scopus)

Abstract

Mixed-linkage β-glucan from barley, with a cellotriosyl to cellotetraosyl ratio of 2.9, was hydrolysed with two endo-1,4-β-glucanases (cellulases) and one non-cellulolytic β-glucanase isolated from Trichoderma reesei. The hydrolysates were precipitated in 90% ethanol and the fragments obtained were further treated with lichenase, followed by analysis of the oligosaccharides released. One of the endo-1,4-β glucanases, Cel 5A (EG II), selectively degraded cellotetraosyl units in the polymer and left the cellotriosyl units unhydrolysed. Blocks of cellotriosyl units were isolated on a larger scale using this enzyme. The isolated blocks were fractionated into four fractions using gel permeation chromatography on Biogel P-6 and the structures of the blocks were analysed by 1H NMR spectroscopy. The fractions essentially contained cellotriosyl units of different sizes with a 3-linked glucose residue at the non-reducing end and a reducing end linked to two 4-linked glucose residues. The results thus indicated that the enzyme could hydrolyse a 4-linked glucose residue next to the 3-linked residue at the non-reducing terminal but left two 4-linked glucose residues at the reducing end. The isolated blocks of cellotriosyl units had a molecular weight distribution that fits a theoretical model based on random blocks of triosyl units in the mixed-linkage β-glucan.
Original languageEnglish
Pages (from-to)233-238
Number of pages6
JournalCarbohydrate Polymers
Volume64
Issue number2
DOIs
Publication statusPublished - 2006
MoE publication typeA1 Journal article-refereed

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Glucans
Glucose
Enzymes
Cellulases
Oligosaccharides
Gel permeation chromatography
Molecular weight distribution
Nuclear magnetic resonance spectroscopy
Polymers
Ethanol

Keywords

  • Barley beta-glucan
  • Cellotriosyl blocks
  • Endo-glucanase

Cite this

Ajithkumar, Anu ; Andersson, Roger ; Siika-aho, Matti ; Tenkanen, Maija ; Åman, Per. / Isolation of cellotriosyl blocks from barley β-glucan with endo-1,4-β-glucanase from Trichoderma reesei. In: Carbohydrate Polymers. 2006 ; Vol. 64, No. 2. pp. 233-238.
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abstract = "Mixed-linkage β-glucan from barley, with a cellotriosyl to cellotetraosyl ratio of 2.9, was hydrolysed with two endo-1,4-β-glucanases (cellulases) and one non-cellulolytic β-glucanase isolated from Trichoderma reesei. The hydrolysates were precipitated in 90{\%} ethanol and the fragments obtained were further treated with lichenase, followed by analysis of the oligosaccharides released. One of the endo-1,4-β glucanases, Cel 5A (EG II), selectively degraded cellotetraosyl units in the polymer and left the cellotriosyl units unhydrolysed. Blocks of cellotriosyl units were isolated on a larger scale using this enzyme. The isolated blocks were fractionated into four fractions using gel permeation chromatography on Biogel P-6 and the structures of the blocks were analysed by 1H NMR spectroscopy. The fractions essentially contained cellotriosyl units of different sizes with a 3-linked glucose residue at the non-reducing end and a reducing end linked to two 4-linked glucose residues. The results thus indicated that the enzyme could hydrolyse a 4-linked glucose residue next to the 3-linked residue at the non-reducing terminal but left two 4-linked glucose residues at the reducing end. The isolated blocks of cellotriosyl units had a molecular weight distribution that fits a theoretical model based on random blocks of triosyl units in the mixed-linkage β-glucan.",
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author = "Anu Ajithkumar and Roger Andersson and Matti Siika-aho and Maija Tenkanen and Per {\AA}man",
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language = "English",
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pages = "233--238",
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Isolation of cellotriosyl blocks from barley β-glucan with endo-1,4-β-glucanase from Trichoderma reesei. / Ajithkumar, Anu (Corresponding Author); Andersson, Roger; Siika-aho, Matti; Tenkanen, Maija; Åman, Per.

In: Carbohydrate Polymers, Vol. 64, No. 2, 2006, p. 233-238.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Isolation of cellotriosyl blocks from barley β-glucan with endo-1,4-β-glucanase from Trichoderma reesei

AU - Ajithkumar, Anu

AU - Andersson, Roger

AU - Siika-aho, Matti

AU - Tenkanen, Maija

AU - Åman, Per

PY - 2006

Y1 - 2006

N2 - Mixed-linkage β-glucan from barley, with a cellotriosyl to cellotetraosyl ratio of 2.9, was hydrolysed with two endo-1,4-β-glucanases (cellulases) and one non-cellulolytic β-glucanase isolated from Trichoderma reesei. The hydrolysates were precipitated in 90% ethanol and the fragments obtained were further treated with lichenase, followed by analysis of the oligosaccharides released. One of the endo-1,4-β glucanases, Cel 5A (EG II), selectively degraded cellotetraosyl units in the polymer and left the cellotriosyl units unhydrolysed. Blocks of cellotriosyl units were isolated on a larger scale using this enzyme. The isolated blocks were fractionated into four fractions using gel permeation chromatography on Biogel P-6 and the structures of the blocks were analysed by 1H NMR spectroscopy. The fractions essentially contained cellotriosyl units of different sizes with a 3-linked glucose residue at the non-reducing end and a reducing end linked to two 4-linked glucose residues. The results thus indicated that the enzyme could hydrolyse a 4-linked glucose residue next to the 3-linked residue at the non-reducing terminal but left two 4-linked glucose residues at the reducing end. The isolated blocks of cellotriosyl units had a molecular weight distribution that fits a theoretical model based on random blocks of triosyl units in the mixed-linkage β-glucan.

AB - Mixed-linkage β-glucan from barley, with a cellotriosyl to cellotetraosyl ratio of 2.9, was hydrolysed with two endo-1,4-β-glucanases (cellulases) and one non-cellulolytic β-glucanase isolated from Trichoderma reesei. The hydrolysates were precipitated in 90% ethanol and the fragments obtained were further treated with lichenase, followed by analysis of the oligosaccharides released. One of the endo-1,4-β glucanases, Cel 5A (EG II), selectively degraded cellotetraosyl units in the polymer and left the cellotriosyl units unhydrolysed. Blocks of cellotriosyl units were isolated on a larger scale using this enzyme. The isolated blocks were fractionated into four fractions using gel permeation chromatography on Biogel P-6 and the structures of the blocks were analysed by 1H NMR spectroscopy. The fractions essentially contained cellotriosyl units of different sizes with a 3-linked glucose residue at the non-reducing end and a reducing end linked to two 4-linked glucose residues. The results thus indicated that the enzyme could hydrolyse a 4-linked glucose residue next to the 3-linked residue at the non-reducing terminal but left two 4-linked glucose residues at the reducing end. The isolated blocks of cellotriosyl units had a molecular weight distribution that fits a theoretical model based on random blocks of triosyl units in the mixed-linkage β-glucan.

KW - Barley beta-glucan

KW - Cellotriosyl blocks

KW - Endo-glucanase

U2 - 10.1016/j.carbpol.2005.11.033

DO - 10.1016/j.carbpol.2005.11.033

M3 - Article

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SP - 233

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JO - Carbohydrate Polymers

JF - Carbohydrate Polymers

SN - 0144-8617

IS - 2

ER -