Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei

Vesa Harjunpää, Anita Teleman, Anu Koivula, Laura Ruohonen, Tuula Teeri, Olle Teleman, Torbjörn Drakenberg

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

The hydrolysis of soluble cello‐oligosaccharides, with a degree of polymerisation of 4–6, catalysed by cellobiohydrolase II from Trichoderma reesei was studied using 1H‐NMR spectroscopy and HPLC. The experimental progress curves were analysed by fitting numerically integrated kinetic equations, which provided cleavage patterns and kinetic constants for each oligosaccharide. This analysis procedure accounts for product inhibition and avoids the initial slope approximation. No glucose was detected at the beginning of the reaction indicating that only the internal glycosidic linkages are attacked. For cellotetraose only the second glycosidic linkage was cleaved. For cellopentaose and cellohexaose the second and the third glycosidic linkage from the non‐reducing end were cleaved with approximately equal probability. The degradation rates of these cello‐oligosaccharides, 1–12 s−1 at 27°C, are about 10–100 times faster than for the 4‐methylumbelliferyl substituted analogs or for cellotriose. No intermediate products larger than cellotriose were released. The degradation rate for cellotetraose were higher than its off‐rate, which accounts for the processive degradation of cellohexaose. A high cellohexaose/enzyme ratio caused slow reversible inactivation of the enzyme.
Original languageEnglish
Pages (from-to)584-591
Number of pages8
JournalEuropean Journal of Biochemistry
Volume240
Issue number3
DOIs
Publication statusPublished - 1996
MoE publication typeA1 Journal article-refereed

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Cellulose 1,4-beta-Cellobiosidase
Trichoderma
Oligosaccharides
Hydrolysis
Degradation
Kinetics
Enzymes
Polymerization
Spectrum Analysis
High Pressure Liquid Chromatography
Spectroscopy
Glucose
cellohexaose
cellotetraose

Cite this

Harjunpää, Vesa ; Teleman, Anita ; Koivula, Anu ; Ruohonen, Laura ; Teeri, Tuula ; Teleman, Olle ; Drakenberg, Torbjörn. / Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei. In: European Journal of Biochemistry. 1996 ; Vol. 240, No. 3. pp. 584-591.
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title = "Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei",
abstract = "The hydrolysis of soluble cello‐oligosaccharides, with a degree of polymerisation of 4–6, catalysed by cellobiohydrolase II from Trichoderma reesei was studied using 1H‐NMR spectroscopy and HPLC. The experimental progress curves were analysed by fitting numerically integrated kinetic equations, which provided cleavage patterns and kinetic constants for each oligosaccharide. This analysis procedure accounts for product inhibition and avoids the initial slope approximation. No glucose was detected at the beginning of the reaction indicating that only the internal glycosidic linkages are attacked. For cellotetraose only the second glycosidic linkage was cleaved. For cellopentaose and cellohexaose the second and the third glycosidic linkage from the non‐reducing end were cleaved with approximately equal probability. The degradation rates of these cello‐oligosaccharides, 1–12 s−1 at 27°C, are about 10–100 times faster than for the 4‐methylumbelliferyl substituted analogs or for cellotriose. No intermediate products larger than cellotriose were released. The degradation rate for cellotetraose were higher than its off‐rate, which accounts for the processive degradation of cellohexaose. A high cellohexaose/enzyme ratio caused slow reversible inactivation of the enzyme.",
author = "Vesa Harjunp{\"a}{\"a} and Anita Teleman and Anu Koivula and Laura Ruohonen and Tuula Teeri and Olle Teleman and Torbj{\"o}rn Drakenberg",
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Harjunpää, V, Teleman, A, Koivula, A, Ruohonen, L, Teeri, T, Teleman, O & Drakenberg, T 1996, 'Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei', European Journal of Biochemistry, vol. 240, no. 3, pp. 584-591. https://doi.org/10.1111/j.1432-1033.1996.0584h.x

Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei. / Harjunpää, Vesa; Teleman, Anita; Koivula, Anu; Ruohonen, Laura; Teeri, Tuula; Teleman, Olle; Drakenberg, Torbjörn.

In: European Journal of Biochemistry, Vol. 240, No. 3, 1996, p. 584-591.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Cello-oligosaccharide hydrolysis by cellobiohydrolase II from Trichoderma reesei

AU - Harjunpää, Vesa

AU - Teleman, Anita

AU - Koivula, Anu

AU - Ruohonen, Laura

AU - Teeri, Tuula

AU - Teleman, Olle

AU - Drakenberg, Torbjörn

N1 - Project code: BEL4319

PY - 1996

Y1 - 1996

N2 - The hydrolysis of soluble cello‐oligosaccharides, with a degree of polymerisation of 4–6, catalysed by cellobiohydrolase II from Trichoderma reesei was studied using 1H‐NMR spectroscopy and HPLC. The experimental progress curves were analysed by fitting numerically integrated kinetic equations, which provided cleavage patterns and kinetic constants for each oligosaccharide. This analysis procedure accounts for product inhibition and avoids the initial slope approximation. No glucose was detected at the beginning of the reaction indicating that only the internal glycosidic linkages are attacked. For cellotetraose only the second glycosidic linkage was cleaved. For cellopentaose and cellohexaose the second and the third glycosidic linkage from the non‐reducing end were cleaved with approximately equal probability. The degradation rates of these cello‐oligosaccharides, 1–12 s−1 at 27°C, are about 10–100 times faster than for the 4‐methylumbelliferyl substituted analogs or for cellotriose. No intermediate products larger than cellotriose were released. The degradation rate for cellotetraose were higher than its off‐rate, which accounts for the processive degradation of cellohexaose. A high cellohexaose/enzyme ratio caused slow reversible inactivation of the enzyme.

AB - The hydrolysis of soluble cello‐oligosaccharides, with a degree of polymerisation of 4–6, catalysed by cellobiohydrolase II from Trichoderma reesei was studied using 1H‐NMR spectroscopy and HPLC. The experimental progress curves were analysed by fitting numerically integrated kinetic equations, which provided cleavage patterns and kinetic constants for each oligosaccharide. This analysis procedure accounts for product inhibition and avoids the initial slope approximation. No glucose was detected at the beginning of the reaction indicating that only the internal glycosidic linkages are attacked. For cellotetraose only the second glycosidic linkage was cleaved. For cellopentaose and cellohexaose the second and the third glycosidic linkage from the non‐reducing end were cleaved with approximately equal probability. The degradation rates of these cello‐oligosaccharides, 1–12 s−1 at 27°C, are about 10–100 times faster than for the 4‐methylumbelliferyl substituted analogs or for cellotriose. No intermediate products larger than cellotriose were released. The degradation rate for cellotetraose were higher than its off‐rate, which accounts for the processive degradation of cellohexaose. A high cellohexaose/enzyme ratio caused slow reversible inactivation of the enzyme.

U2 - 10.1111/j.1432-1033.1996.0584h.x

DO - 10.1111/j.1432-1033.1996.0584h.x

M3 - Article

VL - 240

SP - 584

EP - 591

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 3

ER -