1H, 13C, 15N backbone and IVL methyl group resonance assignment of the fungal β-glucosidase from Trichoderma reesei

Eleni Makraki; Marta G. Carneiro; Alex Heyam; A. B. Eiso; Gregg Siegal; Roderick E. Hubbard

doi:10.1007/s12104-020-09959-2

¹H, ¹³C, ¹⁵N backbone and IVL methyl group resonance assignment of the fungal β-glucosidase from Trichoderma reesei

Eleni Makraki, Marta G. Carneiro, Alex Heyam, A. B. Eiso, Gregg Siegal, Roderick E. Hubbard^*

^*Corresponding author for this work

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

β-glucosidases have received considerable attention due to their essential role in bioethanol production from lignocellulosic biomass. β-glucosidase can hydrolyse cellobiose in cellulose degradation and its low activity has been considered as one of the main limiting steps in the process. Large-scale conversions of cellulose therefore require high enzyme concentration which increases the cost. β-glucosidases with improved activity and thermostability are therefore of great commercial interest. The fungus Trichoderma reseei expresses thermostable cellulolytic enzymes which have been widely studied as attractive targets for industrial applications. Genetically modified β-glucosidases from Trichoderma reseei have been recently commercialised. We have developed an approach in which screening of low molecular weight molecules (fragments) identifies compounds that increase enzyme activity and are currently characterizing fragment-based activators of TrBgl2. A structural analysis of the 55 kDa apo form of TrBgl2 revealed a classical (α/β)₈-TIM barrel fold. In the present study we present a partial assignment of backbone chemical shifts, along with those of the Ile (I)-Val (V)-Leu (L) methyl groups of TrBgl2. These data will be used to characterize the interaction of TrBgl2 with the small molecule activators.

Original language	English
Pages (from-to)	265-268
Number of pages	4
Journal	Biomolecular NMR Assignments
Volume	14
Issue number	2
Early online date	19 Jun 2020
DOIs	https://doi.org/10.1007/s12104-020-09959-2
Publication status	Published - Oct 2020

Funding

This work was supported by the European Union\u2019s Horizon2020 MSCA Programme under Grant agreement 675899 (FRAGNET); research in the group of R.E.H. was additionally supported by Research Grants from the BBSRC (BB/N008332/1) and institutional infrastructure support from funds provided by the Welcome Trust and EPSRC. Acknowledgements EM is very grateful to Michael Plevin (University of York) for his help and support in sample preparation. This work was supported by the European Union?s Horizon2020 MSCA Programme under grant agreement 675899 (FRAGNET); research in the group of R.E.H. was additionally supported by research Grants from the BBSRC (BB/N008332/1) and institutional infrastructure support from funds provided by the Wellcome Trust and EPSRC. EM is very grateful to Michael Plevin (University of York) for his help and support in sample preparation. This work was supported by the European Union\u2019s Horizon2020 MSCA Programme under grant agreement 675899 (FRAGNET); research in the group of R.E.H. was additionally supported by research Grants from the BBSRC (BB/N008332/1) and institutional infrastructure support from funds provided by the Wellcome Trust and EPSRC.

Funders	Funder number
European Union’s Horizon2020 MSCA
European Union?s Horizon2020 MSCA
Wellcome Trust
Engineering and Physical Sciences Research Council
Horizon 2020 Framework Programme	675899
Biotechnology and Biological Sciences Research Council	BB/N008332/1

Keywords

Enzyme activators
Resonance assignment
TrBgl2
β-glucosidase

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Cite this

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title = "1H, 13C, 15N backbone and IVL methyl group resonance assignment of the fungal β-glucosidase from Trichoderma reesei",

abstract = "β-glucosidases have received considerable attention due to their essential role in bioethanol production from lignocellulosic biomass. β-glucosidase can hydrolyse cellobiose in cellulose degradation and its low activity has been considered as one of the main limiting steps in the process. Large-scale conversions of cellulose therefore require high enzyme concentration which increases the cost. β-glucosidases with improved activity and thermostability are therefore of great commercial interest. The fungus Trichoderma reseei expresses thermostable cellulolytic enzymes which have been widely studied as attractive targets for industrial applications. Genetically modified β-glucosidases from Trichoderma reseei have been recently commercialised. We have developed an approach in which screening of low molecular weight molecules (fragments) identifies compounds that increase enzyme activity and are currently characterizing fragment-based activators of TrBgl2. A structural analysis of the 55 kDa apo form of TrBgl2 revealed a classical (α/β)8-TIM barrel fold. In the present study we present a partial assignment of backbone chemical shifts, along with those of the Ile (I)-Val (V)-Leu (L) methyl groups of TrBgl2. These data will be used to characterize the interaction of TrBgl2 with the small molecule activators.",

keywords = "Enzyme activators, Resonance assignment, TrBgl2, β-glucosidase",

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AU - Carneiro, Marta G.

AU - Heyam, Alex

AU - Eiso, A. B.

AU - Siegal, Gregg

AU - Hubbard, Roderick E.

PY - 2020/10

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