Widely different off rates of two closely related cellulose-binding domains from Trichoderma reesei

Geraldine Carrard (Corresponding Author), Markus Linder

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

The filamentous fungus Trichoderma reesei produces two cellobiohydrolases (CBHI and CBHII). These, like most other cellulose‐degrading enzymes, have a modular structure consisting of a catalytic domain linked to a cellulose‐binding domain (CBD). The isolated catalytic domains bind poorly to cellulose and have a much lower activity towards cellulose than the intact enzymes. For the CBDs, no function other than binding to cellulose has been found. We have previously described the reversibility and exchange rate for the binding of the CBD of CBHI to cellulose. In this work, we studied the binding of the CBD of CBHII and showed that it differs markedly from the behaviour of that of CBHI. The apparent binding affinities were similar, but the CBD of CBHII could not be dissociated from cellulose by buffer dilution and did not show a measurable exchange rate. However, desorption could be triggered by shifting the temperature. The CBD of CBHII bound reversibly to chitin. Two variants of the CBHII CBD were made, in which point mutations increased its similarity to the CBD of CBHI. Both variants were found to bind reversibly to cellulose.
Original languageEnglish
Pages (from-to)637-643
Number of pages7
JournalEuropean Journal of Biochemistry
Volume262
Issue number3
DOIs
Publication statusPublished - 1999
MoE publication typeA1 Journal article-refereed

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Trichoderma
Cellulose
Catalytic Domain
Cellulose 1,4-beta-Cellobiosidase
Chitin
Enzymes
Fungi
Point Mutation
Dilution
Desorption
Ion exchange
Buffers
Temperature

Cite this

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title = "Widely different off rates of two closely related cellulose-binding domains from Trichoderma reesei",
abstract = "The filamentous fungus Trichoderma reesei produces two cellobiohydrolases (CBHI and CBHII). These, like most other cellulose‐degrading enzymes, have a modular structure consisting of a catalytic domain linked to a cellulose‐binding domain (CBD). The isolated catalytic domains bind poorly to cellulose and have a much lower activity towards cellulose than the intact enzymes. For the CBDs, no function other than binding to cellulose has been found. We have previously described the reversibility and exchange rate for the binding of the CBD of CBHI to cellulose. In this work, we studied the binding of the CBD of CBHII and showed that it differs markedly from the behaviour of that of CBHI. The apparent binding affinities were similar, but the CBD of CBHII could not be dissociated from cellulose by buffer dilution and did not show a measurable exchange rate. However, desorption could be triggered by shifting the temperature. The CBD of CBHII bound reversibly to chitin. Two variants of the CBHII CBD were made, in which point mutations increased its similarity to the CBD of CBHI. Both variants were found to bind reversibly to cellulose.",
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Widely different off rates of two closely related cellulose-binding domains from Trichoderma reesei. / Carrard, Geraldine (Corresponding Author); Linder, Markus.

In: European Journal of Biochemistry, Vol. 262, No. 3, 1999, p. 637-643.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Widely different off rates of two closely related cellulose-binding domains from Trichoderma reesei

AU - Carrard, Geraldine

AU - Linder, Markus

PY - 1999

Y1 - 1999

N2 - The filamentous fungus Trichoderma reesei produces two cellobiohydrolases (CBHI and CBHII). These, like most other cellulose‐degrading enzymes, have a modular structure consisting of a catalytic domain linked to a cellulose‐binding domain (CBD). The isolated catalytic domains bind poorly to cellulose and have a much lower activity towards cellulose than the intact enzymes. For the CBDs, no function other than binding to cellulose has been found. We have previously described the reversibility and exchange rate for the binding of the CBD of CBHI to cellulose. In this work, we studied the binding of the CBD of CBHII and showed that it differs markedly from the behaviour of that of CBHI. The apparent binding affinities were similar, but the CBD of CBHII could not be dissociated from cellulose by buffer dilution and did not show a measurable exchange rate. However, desorption could be triggered by shifting the temperature. The CBD of CBHII bound reversibly to chitin. Two variants of the CBHII CBD were made, in which point mutations increased its similarity to the CBD of CBHI. Both variants were found to bind reversibly to cellulose.

AB - The filamentous fungus Trichoderma reesei produces two cellobiohydrolases (CBHI and CBHII). These, like most other cellulose‐degrading enzymes, have a modular structure consisting of a catalytic domain linked to a cellulose‐binding domain (CBD). The isolated catalytic domains bind poorly to cellulose and have a much lower activity towards cellulose than the intact enzymes. For the CBDs, no function other than binding to cellulose has been found. We have previously described the reversibility and exchange rate for the binding of the CBD of CBHI to cellulose. In this work, we studied the binding of the CBD of CBHII and showed that it differs markedly from the behaviour of that of CBHI. The apparent binding affinities were similar, but the CBD of CBHII could not be dissociated from cellulose by buffer dilution and did not show a measurable exchange rate. However, desorption could be triggered by shifting the temperature. The CBD of CBHII bound reversibly to chitin. Two variants of the CBHII CBD were made, in which point mutations increased its similarity to the CBD of CBHI. Both variants were found to bind reversibly to cellulose.

U2 - 10.1046/j.1432-1327.1999.00455.x

DO - 10.1046/j.1432-1327.1999.00455.x

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JF - FEBS Journal

SN - 1742-464X

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