A biochemical comparison of fungal GH6 cellobiohydrolases

Stefan Jarl Christensen, Kristian Bertel Rømer Mørkeberg Krogh, Nikolaj Spodsberg, Kim Borch, Peter Westh*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Cellobiohydrolases (CBHs) from glycoside hydrolase family 6 (GH6) make up an important part of the secretome in many cellulolytic fungi. They are also of technical interest, particularly because they are part of the enzyme cocktails that are used for the industrial breakdown of lignocellulosic biomass. Nevertheless, functional studies of GH6 CBHs are scarce and focused on a few model enzymes. To elucidate functional breadth among GH6 CBHs, we conducted a comparative biochemical study of seven GH6 CBHs originating from fungi living in different habitats, in addition to one enzyme variant. The enzyme sequences were investigated by phylogenetic analyses to ensure that they were not closely related phylogenetically. The selected enzymes were all heterologously expressed in Aspergillus oryzae, purified and thoroughly characterized biochemically. This approach allowed direct comparisons of functional data, and the results revealed substantial variability. For example, the adsorption capacity on cellulose spanned two orders of magnitude and kinetic parameters, derived from two independent steady-state methods also varied significantly. While the different functional parameters covered wide ranges, they were not independent since they changed in parallel between two poles. One pole was characterized by strong substrate interactions, high adsorption capacity and low turnover number while the other showed weak substrate interactions, poor adsorption and high turnover. The investigated enzymes essentially defined a continuum between these two opposites, and this scaling of functional parameters raises interesting questions regarding functional plasticity and evolution of GH6 CBHs.

Original languageEnglish
JournalBiochemical Journal
Volume476
Issue number15
Pages (from-to)2157-2172
Number of pages16
ISSN0264-6021
DOIs
Publication statusPublished - 2019

Cite this

Christensen, S. J., Krogh, K. B. R. M., Spodsberg, N., Borch, K., & Westh, P. (2019). A biochemical comparison of fungal GH6 cellobiohydrolases. Biochemical Journal, 476(15), 2157-2172. https://doi.org/10.1042/BCJ20190185
Christensen, Stefan Jarl ; Krogh, Kristian Bertel Rømer Mørkeberg ; Spodsberg, Nikolaj ; Borch, Kim ; Westh, Peter. / A biochemical comparison of fungal GH6 cellobiohydrolases. In: Biochemical Journal. 2019 ; Vol. 476, No. 15. pp. 2157-2172.
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abstract = "Cellobiohydrolases (CBHs) from glycoside hydrolase family 6 (GH6) make up an important part of the secretome in many cellulolytic fungi. They are also of technical interest, particularly because they are part of the enzyme cocktails that are used for the industrial breakdown of lignocellulosic biomass. Nevertheless, functional studies of GH6 CBHs are scarce and focused on a few model enzymes. To elucidate functional breadth among GH6 CBHs, we conducted a comparative biochemical study of seven GH6 CBHs originating from fungi living in different habitats, in addition to one enzyme variant. The enzyme sequences were investigated by phylogenetic analyses to ensure that they were not closely related phylogenetically. The selected enzymes were all heterologously expressed in Aspergillus oryzae, purified and thoroughly characterized biochemically. This approach allowed direct comparisons of functional data, and the results revealed substantial variability. For example, the adsorption capacity on cellulose spanned two orders of magnitude and kinetic parameters, derived from two independent steady-state methods also varied significantly. While the different functional parameters covered wide ranges, they were not independent since they changed in parallel between two poles. One pole was characterized by strong substrate interactions, high adsorption capacity and low turnover number while the other showed weak substrate interactions, poor adsorption and high turnover. The investigated enzymes essentially defined a continuum between these two opposites, and this scaling of functional parameters raises interesting questions regarding functional plasticity and evolution of GH6 CBHs.",
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Christensen, SJ, Krogh, KBRM, Spodsberg, N, Borch, K & Westh, P 2019, 'A biochemical comparison of fungal GH6 cellobiohydrolases', Biochemical Journal, vol. 476, no. 15, pp. 2157-2172. https://doi.org/10.1042/BCJ20190185

A biochemical comparison of fungal GH6 cellobiohydrolases. / Christensen, Stefan Jarl; Krogh, Kristian Bertel Rømer Mørkeberg; Spodsberg, Nikolaj; Borch, Kim; Westh, Peter.

In: Biochemical Journal, Vol. 476, No. 15, 2019, p. 2157-2172.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - A biochemical comparison of fungal GH6 cellobiohydrolases

AU - Christensen, Stefan Jarl

AU - Krogh, Kristian Bertel Rømer Mørkeberg

AU - Spodsberg, Nikolaj

AU - Borch, Kim

AU - Westh, Peter

PY - 2019

Y1 - 2019

N2 - Cellobiohydrolases (CBHs) from glycoside hydrolase family 6 (GH6) make up an important part of the secretome in many cellulolytic fungi. They are also of technical interest, particularly because they are part of the enzyme cocktails that are used for the industrial breakdown of lignocellulosic biomass. Nevertheless, functional studies of GH6 CBHs are scarce and focused on a few model enzymes. To elucidate functional breadth among GH6 CBHs, we conducted a comparative biochemical study of seven GH6 CBHs originating from fungi living in different habitats, in addition to one enzyme variant. The enzyme sequences were investigated by phylogenetic analyses to ensure that they were not closely related phylogenetically. The selected enzymes were all heterologously expressed in Aspergillus oryzae, purified and thoroughly characterized biochemically. This approach allowed direct comparisons of functional data, and the results revealed substantial variability. For example, the adsorption capacity on cellulose spanned two orders of magnitude and kinetic parameters, derived from two independent steady-state methods also varied significantly. While the different functional parameters covered wide ranges, they were not independent since they changed in parallel between two poles. One pole was characterized by strong substrate interactions, high adsorption capacity and low turnover number while the other showed weak substrate interactions, poor adsorption and high turnover. The investigated enzymes essentially defined a continuum between these two opposites, and this scaling of functional parameters raises interesting questions regarding functional plasticity and evolution of GH6 CBHs.

AB - Cellobiohydrolases (CBHs) from glycoside hydrolase family 6 (GH6) make up an important part of the secretome in many cellulolytic fungi. They are also of technical interest, particularly because they are part of the enzyme cocktails that are used for the industrial breakdown of lignocellulosic biomass. Nevertheless, functional studies of GH6 CBHs are scarce and focused on a few model enzymes. To elucidate functional breadth among GH6 CBHs, we conducted a comparative biochemical study of seven GH6 CBHs originating from fungi living in different habitats, in addition to one enzyme variant. The enzyme sequences were investigated by phylogenetic analyses to ensure that they were not closely related phylogenetically. The selected enzymes were all heterologously expressed in Aspergillus oryzae, purified and thoroughly characterized biochemically. This approach allowed direct comparisons of functional data, and the results revealed substantial variability. For example, the adsorption capacity on cellulose spanned two orders of magnitude and kinetic parameters, derived from two independent steady-state methods also varied significantly. While the different functional parameters covered wide ranges, they were not independent since they changed in parallel between two poles. One pole was characterized by strong substrate interactions, high adsorption capacity and low turnover number while the other showed weak substrate interactions, poor adsorption and high turnover. The investigated enzymes essentially defined a continuum between these two opposites, and this scaling of functional parameters raises interesting questions regarding functional plasticity and evolution of GH6 CBHs.

U2 - 10.1042/BCJ20190185

DO - 10.1042/BCJ20190185

M3 - Journal article

VL - 476

SP - 2157

EP - 2172

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 15

ER -