A comparative study of two retaining enzymes of Trichoderma reesei

Transglycosylation of oligosaccharides catalysed by the cellobiohydrolase I, Cel7A, and the β-mannanase, Man5A

Vesa Harjunpää (Corresponding Author), Jari Helin, Anu Koivula, Matti Siika-aho, Torbjörn Drakenberg

Research output: Contribution to journalArticleScientificpeer-review

34 Citations (Scopus)

Abstract

HPLC, MALDI-TOF MS and NMR spectroscopy were used to investigate the hydrolysis of cello- and manno-oligosaccharides by Cel7A and Man5A from Trichoderma reesei. The experimental progress curves were analysed by fitting the numerically integrated kinetic equations, which provided cleavage patterns for oligosaccharides. This data evaluation procedure accounts for product inhibition and avoids the initial slope approximation. In addition, a transglycosylation step had to be included in the model to reproduce the experimental progress curves. For the hydrolysis of manno-oligosaccharides, Man4–6, by Man5A no mannose was detected at the beginning of the reaction showing that only the internal linkages are hydrolysed. For cellotriose and cellotetraose hydrolysis by Cel7A, the main product is cellobiose and glucose is released from the non-reducing end of the substrate. Intermediary products longer than the substrates were detected by MALDI-TOF MS when oligosaccharides (Glc4–6 or Man4–6) were hydrolysed by either Cel7A or Man5A. Interestingly, two distinct transglycosylation pathways could be observed. Cel7A produced intermediates that are one unit longer than the substrate, whereas Man5A produced intermediates that are two units longer than the substrate.
Original languageEnglish
Pages (from-to)149-153
Number of pages5
JournalFEBS Letters
Volume443
Issue number2
DOIs
Publication statusPublished - 1999
MoE publication typeA1 Journal article-refereed

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Cellulose 1,4-beta-Cellobiosidase
Trichoderma
Oligosaccharides
Enzymes
Hydrolysis
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Substrates
Cellobiose
Mannose
Nuclear magnetic resonance spectroscopy
Magnetic Resonance Spectroscopy
High Pressure Liquid Chromatography
Glucose
Kinetics

Keywords

  • cellulace
  • Mannanase
  • Oligosaccharide
  • Transglycosylation
  • Trichoderma reesei

Cite this

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title = "A comparative study of two retaining enzymes of Trichoderma reesei: Transglycosylation of oligosaccharides catalysed by the cellobiohydrolase I, Cel7A, and the β-mannanase, Man5A",
abstract = "HPLC, MALDI-TOF MS and NMR spectroscopy were used to investigate the hydrolysis of cello- and manno-oligosaccharides by Cel7A and Man5A from Trichoderma reesei. The experimental progress curves were analysed by fitting the numerically integrated kinetic equations, which provided cleavage patterns for oligosaccharides. This data evaluation procedure accounts for product inhibition and avoids the initial slope approximation. In addition, a transglycosylation step had to be included in the model to reproduce the experimental progress curves. For the hydrolysis of manno-oligosaccharides, Man4–6, by Man5A no mannose was detected at the beginning of the reaction showing that only the internal linkages are hydrolysed. For cellotriose and cellotetraose hydrolysis by Cel7A, the main product is cellobiose and glucose is released from the non-reducing end of the substrate. Intermediary products longer than the substrates were detected by MALDI-TOF MS when oligosaccharides (Glc4–6 or Man4–6) were hydrolysed by either Cel7A or Man5A. Interestingly, two distinct transglycosylation pathways could be observed. Cel7A produced intermediates that are one unit longer than the substrate, whereas Man5A produced intermediates that are two units longer than the substrate.",
keywords = "cellulace, Mannanase, Oligosaccharide, Transglycosylation, Trichoderma reesei",
author = "Vesa Harjunp{\"a}{\"a} and Jari Helin and Anu Koivula and Matti Siika-aho and Torbj{\"o}rn Drakenberg",
year = "1999",
doi = "10.1016/S0014-5793(98)01692-5",
language = "English",
volume = "443",
pages = "149--153",
journal = "FEBS Letters",
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}

A comparative study of two retaining enzymes of Trichoderma reesei : Transglycosylation of oligosaccharides catalysed by the cellobiohydrolase I, Cel7A, and the β-mannanase, Man5A. / Harjunpää, Vesa (Corresponding Author); Helin, Jari; Koivula, Anu; Siika-aho, Matti; Drakenberg, Torbjörn.

In: FEBS Letters, Vol. 443, No. 2, 1999, p. 149-153.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - A comparative study of two retaining enzymes of Trichoderma reesei

T2 - Transglycosylation of oligosaccharides catalysed by the cellobiohydrolase I, Cel7A, and the β-mannanase, Man5A

AU - Harjunpää, Vesa

AU - Helin, Jari

AU - Koivula, Anu

AU - Siika-aho, Matti

AU - Drakenberg, Torbjörn

PY - 1999

Y1 - 1999

N2 - HPLC, MALDI-TOF MS and NMR spectroscopy were used to investigate the hydrolysis of cello- and manno-oligosaccharides by Cel7A and Man5A from Trichoderma reesei. The experimental progress curves were analysed by fitting the numerically integrated kinetic equations, which provided cleavage patterns for oligosaccharides. This data evaluation procedure accounts for product inhibition and avoids the initial slope approximation. In addition, a transglycosylation step had to be included in the model to reproduce the experimental progress curves. For the hydrolysis of manno-oligosaccharides, Man4–6, by Man5A no mannose was detected at the beginning of the reaction showing that only the internal linkages are hydrolysed. For cellotriose and cellotetraose hydrolysis by Cel7A, the main product is cellobiose and glucose is released from the non-reducing end of the substrate. Intermediary products longer than the substrates were detected by MALDI-TOF MS when oligosaccharides (Glc4–6 or Man4–6) were hydrolysed by either Cel7A or Man5A. Interestingly, two distinct transglycosylation pathways could be observed. Cel7A produced intermediates that are one unit longer than the substrate, whereas Man5A produced intermediates that are two units longer than the substrate.

AB - HPLC, MALDI-TOF MS and NMR spectroscopy were used to investigate the hydrolysis of cello- and manno-oligosaccharides by Cel7A and Man5A from Trichoderma reesei. The experimental progress curves were analysed by fitting the numerically integrated kinetic equations, which provided cleavage patterns for oligosaccharides. This data evaluation procedure accounts for product inhibition and avoids the initial slope approximation. In addition, a transglycosylation step had to be included in the model to reproduce the experimental progress curves. For the hydrolysis of manno-oligosaccharides, Man4–6, by Man5A no mannose was detected at the beginning of the reaction showing that only the internal linkages are hydrolysed. For cellotriose and cellotetraose hydrolysis by Cel7A, the main product is cellobiose and glucose is released from the non-reducing end of the substrate. Intermediary products longer than the substrates were detected by MALDI-TOF MS when oligosaccharides (Glc4–6 or Man4–6) were hydrolysed by either Cel7A or Man5A. Interestingly, two distinct transglycosylation pathways could be observed. Cel7A produced intermediates that are one unit longer than the substrate, whereas Man5A produced intermediates that are two units longer than the substrate.

KW - cellulace

KW - Mannanase

KW - Oligosaccharide

KW - Transglycosylation

KW - Trichoderma reesei

U2 - 10.1016/S0014-5793(98)01692-5

DO - 10.1016/S0014-5793(98)01692-5

M3 - Article

VL - 443

SP - 149

EP - 153

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 2

ER -