Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A and Cel7B from H. jecorina.

Vanessa de Oliveira Arnoldi Pellegrini, Nina Lei, Madhuri Kysaram, Johan Pelck Olsen, Silke Flindt Badino, Michael Skovbo Windahl, Francieli Colussi, Nicolaj Cruys-Bagger, Kim Borch, Peter Westh

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Adsorption of cellulases on the cellulose surface is an integral part of the catalytic mechanism, and a detailed description of the adsorption process is therefore required for a fundamental understanding of this industrially important class of enzymes. However, the mode of adsorption has proven intricate, and several key questions remain open. Perhaps most notably it is not clear whether the adsorbed enzyme is in dynamic equilibrium with the free population or irreversibly associated with no or slow dissociation. To address this, we have systematically investigated adsorption reversibility for two cellobiohydrolases (Cel7A and Cel6A) and one endoglucanase (Cel7B) on four types of pure cellulose substrates. Specifically, we monitored dilution-induced release of adsorbed enzyme in samples that had previously been brought to a steady state (constant concentration of free enzyme). In simple dilution experiments (without centrifugation), the results consistently showed full reversibility. In contrast to this, resuspension of enzyme–substrate pellets separated by centrifugation showed extensive irreversibility. We conclude that these enzymes are in a dynamic equilibrium between free and adsorbed states but suggest that changes in the physical properties of cellulose caused by compaction of the pellet hampers subsequent release of adsorbed enzyme. This latter effect may be pertinent to both previous controversies in the literature on adsorption reversibility and the development of enzyme recycling protocols in the biomass industry
OriginalsprogEngelsk
TidsskriftLangmuir
Vol/bind30
Udgave nummer42
Sider (fra-til)12602-12609
ISSN0743-7463
DOI
StatusUdgivet - 2014

Citer dette

Pellegrini, Vanessa de Oliveira Arnoldi ; Lei, Nina ; Kysaram, Madhuri ; Olsen, Johan Pelck ; Badino, Silke Flindt ; Windahl, Michael Skovbo ; Colussi, Francieli ; Cruys-Bagger, Nicolaj ; Borch, Kim ; Westh, Peter. / Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A and Cel7B from H. jecorina. I: Langmuir. 2014 ; Bind 30, Nr. 42. s. 12602-12609.
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title = "Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A and Cel7B from H. jecorina.",
abstract = "Adsorption of cellulases on the cellulose surface is an integral part of the catalytic mechanism, and a detailed description of the adsorption process is therefore required for a fundamental understanding of this industrially important class of enzymes. However, the mode of adsorption has proven intricate, and several key questions remain open. Perhaps most notably it is not clear whether the adsorbed enzyme is in dynamic equilibrium with the free population or irreversibly associated with no or slow dissociation. To address this, we have systematically investigated adsorption reversibility for two cellobiohydrolases (Cel7A and Cel6A) and one endoglucanase (Cel7B) on four types of pure cellulose substrates. Specifically, we monitored dilution-induced release of adsorbed enzyme in samples that had previously been brought to a steady state (constant concentration of free enzyme). In simple dilution experiments (without centrifugation), the results consistently showed full reversibility. In contrast to this, resuspension of enzyme–substrate pellets separated by centrifugation showed extensive irreversibility. We conclude that these enzymes are in a dynamic equilibrium between free and adsorbed states but suggest that changes in the physical properties of cellulose caused by compaction of the pellet hampers subsequent release of adsorbed enzyme. This latter effect may be pertinent to both previous controversies in the literature on adsorption reversibility and the development of enzyme recycling protocols in the biomass industry",
author = "Pellegrini, {Vanessa de Oliveira Arnoldi} and Nina Lei and Madhuri Kysaram and Olsen, {Johan Pelck} and Badino, {Silke Flindt} and Windahl, {Michael Skovbo} and Francieli Colussi and Nicolaj Cruys-Bagger and Kim Borch and Peter Westh",
year = "2014",
doi = "10.1021/la5024423",
language = "English",
volume = "30",
pages = "12602--12609",
journal = "Langmuir",
issn = "0743-7463",
publisher = "American Chemical Society",
number = "42",

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Pellegrini, VDOA, Lei, N, Kysaram, M, Olsen, JP, Badino, SF, Windahl, MS, Colussi, F, Cruys-Bagger, N, Borch, K & Westh, P 2014, 'Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A and Cel7B from H. jecorina.', Langmuir, bind 30, nr. 42, s. 12602-12609. https://doi.org/10.1021/la5024423

Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A and Cel7B from H. jecorina. / Pellegrini, Vanessa de Oliveira Arnoldi; Lei, Nina; Kysaram, Madhuri; Olsen, Johan Pelck; Badino, Silke Flindt; Windahl, Michael Skovbo; Colussi, Francieli; Cruys-Bagger, Nicolaj; Borch, Kim; Westh, Peter.

I: Langmuir, Bind 30, Nr. 42, 2014, s. 12602-12609.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Reversibility of substrate adsorption for the cellulases Cel7A, Cel6A and Cel7B from H. jecorina.

AU - Pellegrini, Vanessa de Oliveira Arnoldi

AU - Lei, Nina

AU - Kysaram, Madhuri

AU - Olsen, Johan Pelck

AU - Badino, Silke Flindt

AU - Windahl, Michael Skovbo

AU - Colussi, Francieli

AU - Cruys-Bagger, Nicolaj

AU - Borch, Kim

AU - Westh, Peter

PY - 2014

Y1 - 2014

N2 - Adsorption of cellulases on the cellulose surface is an integral part of the catalytic mechanism, and a detailed description of the adsorption process is therefore required for a fundamental understanding of this industrially important class of enzymes. However, the mode of adsorption has proven intricate, and several key questions remain open. Perhaps most notably it is not clear whether the adsorbed enzyme is in dynamic equilibrium with the free population or irreversibly associated with no or slow dissociation. To address this, we have systematically investigated adsorption reversibility for two cellobiohydrolases (Cel7A and Cel6A) and one endoglucanase (Cel7B) on four types of pure cellulose substrates. Specifically, we monitored dilution-induced release of adsorbed enzyme in samples that had previously been brought to a steady state (constant concentration of free enzyme). In simple dilution experiments (without centrifugation), the results consistently showed full reversibility. In contrast to this, resuspension of enzyme–substrate pellets separated by centrifugation showed extensive irreversibility. We conclude that these enzymes are in a dynamic equilibrium between free and adsorbed states but suggest that changes in the physical properties of cellulose caused by compaction of the pellet hampers subsequent release of adsorbed enzyme. This latter effect may be pertinent to both previous controversies in the literature on adsorption reversibility and the development of enzyme recycling protocols in the biomass industry

AB - Adsorption of cellulases on the cellulose surface is an integral part of the catalytic mechanism, and a detailed description of the adsorption process is therefore required for a fundamental understanding of this industrially important class of enzymes. However, the mode of adsorption has proven intricate, and several key questions remain open. Perhaps most notably it is not clear whether the adsorbed enzyme is in dynamic equilibrium with the free population or irreversibly associated with no or slow dissociation. To address this, we have systematically investigated adsorption reversibility for two cellobiohydrolases (Cel7A and Cel6A) and one endoglucanase (Cel7B) on four types of pure cellulose substrates. Specifically, we monitored dilution-induced release of adsorbed enzyme in samples that had previously been brought to a steady state (constant concentration of free enzyme). In simple dilution experiments (without centrifugation), the results consistently showed full reversibility. In contrast to this, resuspension of enzyme–substrate pellets separated by centrifugation showed extensive irreversibility. We conclude that these enzymes are in a dynamic equilibrium between free and adsorbed states but suggest that changes in the physical properties of cellulose caused by compaction of the pellet hampers subsequent release of adsorbed enzyme. This latter effect may be pertinent to both previous controversies in the literature on adsorption reversibility and the development of enzyme recycling protocols in the biomass industry

U2 - 10.1021/la5024423

DO - 10.1021/la5024423

M3 - Journal article

VL - 30

SP - 12602

EP - 12609

JO - Langmuir

JF - Langmuir

SN - 0743-7463

IS - 42

ER -