Progress-curve analysis shows that glucose inhibits the cellotriose hydrolysis catalysed by cellobiohydrolase II from Trichoderma reesei

Anita Teleman, Anu Koivula, Tapani Reinikainen, Anne Valkeajärvi, Tuula Teeri, Torbjörn Drakenberg, Olle Teleman (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

NMR spectroscopy and HPLC were used to investigate the hydrolysis of cellotriose by cellobiohydrolase II from Trichoderma reesei. Substrate and product concentrations were followed as a function of time. Progress curves were calculated by forward numerical integration of the full kinetic equations and were fitted to the experimental data. Binding and rate constants were obtained from this fit, whereby no initial slope or Michaelis‐Menten approximation was used. The progress curves from a single experiment sufficed to produce agreement with the Michaelis‐Menten model (eight experiments). The absence of a kinetic isotope effect was proven.

The progress‐curve analysis showed that a simple degradation model cannot describe the experimental time‐courses at substrate concentrations greater than 1 mM. A model containing competitive inhibition from cellobiose as well as non‐competitive inhibition from glucose was developed.
This four‐parameter model accurately reproduces about 1000 experimental data points covering five orders of magnitude in oligosaccharide concentrations. Glucose binding to the enzyme/cellotriose complex retards, in a non‐competitive fashion, cellotriose hydrolysis by at least a factor of 30.
A structural model for the non‐competitive inhibition is discussed. The NMR experiment also produced individual progress curves for the α and β anomers.
The β anomer of cellotriose was degraded 2.5‐times faster than the α anomer.
Original languageEnglish
Pages (from-to)250-258
JournalEuropean Journal of Biochemistry
Volume231
Issue number1
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

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Cellulose 1,4-beta-Cellobiosidase
Trichoderma
Hydrolysis
Cellobiose
Glucose
Structural Models
Oligosaccharides
Isotopes
Magnetic Resonance Spectroscopy
High Pressure Liquid Chromatography
Enzymes
Kinetics
Experiments
Substrates
Nuclear magnetic resonance spectroscopy
Rate constants
Nuclear magnetic resonance
Degradation

Cite this

Teleman, Anita ; Koivula, Anu ; Reinikainen, Tapani ; Valkeajärvi, Anne ; Teeri, Tuula ; Drakenberg, Torbjörn ; Teleman, Olle. / Progress-curve analysis shows that glucose inhibits the cellotriose hydrolysis catalysed by cellobiohydrolase II from Trichoderma reesei. In: European Journal of Biochemistry. 1995 ; Vol. 231, No. 1. pp. 250-258.
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abstract = "NMR spectroscopy and HPLC were used to investigate the hydrolysis of cellotriose by cellobiohydrolase II from Trichoderma reesei. Substrate and product concentrations were followed as a function of time. Progress curves were calculated by forward numerical integration of the full kinetic equations and were fitted to the experimental data. Binding and rate constants were obtained from this fit, whereby no initial slope or Michaelis‐Menten approximation was used. The progress curves from a single experiment sufficed to produce agreement with the Michaelis‐Menten model (eight experiments). The absence of a kinetic isotope effect was proven.The progress‐curve analysis showed that a simple degradation model cannot describe the experimental time‐courses at substrate concentrations greater than 1 mM. A model containing competitive inhibition from cellobiose as well as non‐competitive inhibition from glucose was developed. This four‐parameter model accurately reproduces about 1000 experimental data points covering five orders of magnitude in oligosaccharide concentrations. Glucose binding to the enzyme/cellotriose complex retards, in a non‐competitive fashion, cellotriose hydrolysis by at least a factor of 30. A structural model for the non‐competitive inhibition is discussed. The NMR experiment also produced individual progress curves for the α and β anomers. The β anomer of cellotriose was degraded 2.5‐times faster than the α anomer.",
author = "Anita Teleman and Anu Koivula and Tapani Reinikainen and Anne Valkeaj{\"a}rvi and Tuula Teeri and Torbj{\"o}rn Drakenberg and Olle Teleman",
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Progress-curve analysis shows that glucose inhibits the cellotriose hydrolysis catalysed by cellobiohydrolase II from Trichoderma reesei. / Teleman, Anita; Koivula, Anu; Reinikainen, Tapani; Valkeajärvi, Anne; Teeri, Tuula; Drakenberg, Torbjörn; Teleman, Olle (Corresponding Author).

In: European Journal of Biochemistry, Vol. 231, No. 1, 1995, p. 250-258.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Progress-curve analysis shows that glucose inhibits the cellotriose hydrolysis catalysed by cellobiohydrolase II from Trichoderma reesei

AU - Teleman, Anita

AU - Koivula, Anu

AU - Reinikainen, Tapani

AU - Valkeajärvi, Anne

AU - Teeri, Tuula

AU - Drakenberg, Torbjörn

AU - Teleman, Olle

N1 - Project code: KET3112 Project code: BIO3015

PY - 1995

Y1 - 1995

N2 - NMR spectroscopy and HPLC were used to investigate the hydrolysis of cellotriose by cellobiohydrolase II from Trichoderma reesei. Substrate and product concentrations were followed as a function of time. Progress curves were calculated by forward numerical integration of the full kinetic equations and were fitted to the experimental data. Binding and rate constants were obtained from this fit, whereby no initial slope or Michaelis‐Menten approximation was used. The progress curves from a single experiment sufficed to produce agreement with the Michaelis‐Menten model (eight experiments). The absence of a kinetic isotope effect was proven.The progress‐curve analysis showed that a simple degradation model cannot describe the experimental time‐courses at substrate concentrations greater than 1 mM. A model containing competitive inhibition from cellobiose as well as non‐competitive inhibition from glucose was developed. This four‐parameter model accurately reproduces about 1000 experimental data points covering five orders of magnitude in oligosaccharide concentrations. Glucose binding to the enzyme/cellotriose complex retards, in a non‐competitive fashion, cellotriose hydrolysis by at least a factor of 30. A structural model for the non‐competitive inhibition is discussed. The NMR experiment also produced individual progress curves for the α and β anomers. The β anomer of cellotriose was degraded 2.5‐times faster than the α anomer.

AB - NMR spectroscopy and HPLC were used to investigate the hydrolysis of cellotriose by cellobiohydrolase II from Trichoderma reesei. Substrate and product concentrations were followed as a function of time. Progress curves were calculated by forward numerical integration of the full kinetic equations and were fitted to the experimental data. Binding and rate constants were obtained from this fit, whereby no initial slope or Michaelis‐Menten approximation was used. The progress curves from a single experiment sufficed to produce agreement with the Michaelis‐Menten model (eight experiments). The absence of a kinetic isotope effect was proven.The progress‐curve analysis showed that a simple degradation model cannot describe the experimental time‐courses at substrate concentrations greater than 1 mM. A model containing competitive inhibition from cellobiose as well as non‐competitive inhibition from glucose was developed. This four‐parameter model accurately reproduces about 1000 experimental data points covering five orders of magnitude in oligosaccharide concentrations. Glucose binding to the enzyme/cellotriose complex retards, in a non‐competitive fashion, cellotriose hydrolysis by at least a factor of 30. A structural model for the non‐competitive inhibition is discussed. The NMR experiment also produced individual progress curves for the α and β anomers. The β anomer of cellotriose was degraded 2.5‐times faster than the α anomer.

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DO - 10.1111/j.1432-1033.1995.0250f.x

M3 - Article

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SP - 250

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