Thermoactivation of a cellobiohydrolase

Peter Westh, Kim Borch, Trine Holst Sørensen, Radina Tokin, Jeppe Kari, Silke Flindt Badino, Mafalda A. Cavaleiro, Nanna Sandager Røjel, Stefan Jarl Christensen, Cynthia Segura Vesterager, Corinna Schiano di Cola

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

We have measured activity and substrate affinity of the thermostable cellobiohydrolase, Cel7A, from Rasamsonia emersonii over a broad range of temperatures. For the wild type enzyme, which does not have a Carbohydrate Binding Module (CBM), higher temperature only led to moderately increased activity against cellulose, and we ascribed this to a pronounced, temperature induced desorption of enzyme from the substrate surface. We also tested a "high affinity" variant of R. emersonii Cel7A with a linker and CBM from a related enzyme. At room temperature, the activity of the variant was similar to the wild type, but the variant was more accelerated by temperature and about two-fold faster around 70 °C. This better thermoactivation of the high-affinity variant could not be linked to differences in stability or the catalytic process, but coincided with less desorption as temperature increased. Based on these observations and earlier reports on moderate thermoactivation of cellulases, we suggest that better cellulolytic activity at industrially relevant temperatures may be attained by engineering improved substrate affinity into enzymes that already possess good thermostability.
OriginalsprogEngelsk
TidsskriftBiotechnology and Bioengineering (Print)
Vol/bind115
Udgave nummer4
Sider (fra-til)831-838
Antal sider8
ISSN0006-3592
DOI
StatusUdgivet - jan. 2018

Bibliografisk note

This article has been found as a ’Free Version’ from the Publisher on January 4th 2019. When access to the article closes, please notify rucforsk@ruc.dk

Citer dette

Westh, Peter ; Borch, Kim ; Sørensen, Trine Holst ; Tokin, Radina ; Kari, Jeppe ; Badino, Silke Flindt ; Cavaleiro, Mafalda A. ; Røjel, Nanna Sandager ; Christensen, Stefan Jarl ; Vesterager, Cynthia Segura ; Cola, Corinna Schiano di. / Thermoactivation of a cellobiohydrolase. I: Biotechnology and Bioengineering (Print). 2018 ; Bind 115, Nr. 4. s. 831-838.
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abstract = "We have measured activity and substrate affinity of the thermostable cellobiohydrolase, Cel7A, from Rasamsonia emersonii over a broad range of temperatures. For the wild type enzyme, which does not have a Carbohydrate Binding Module (CBM), higher temperature only led to moderately increased activity against cellulose, and we ascribed this to a pronounced, temperature induced desorption of enzyme from the substrate surface. We also tested a {"}high affinity{"} variant of R. emersonii Cel7A with a linker and CBM from a related enzyme. At room temperature, the activity of the variant was similar to the wild type, but the variant was more accelerated by temperature and about two-fold faster around 70 °C. This better thermoactivation of the high-affinity variant could not be linked to differences in stability or the catalytic process, but coincided with less desorption as temperature increased. Based on these observations and earlier reports on moderate thermoactivation of cellulases, we suggest that better cellulolytic activity at industrially relevant temperatures may be attained by engineering improved substrate affinity into enzymes that already possess good thermostability.",
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Thermoactivation of a cellobiohydrolase. / Westh, Peter; Borch, Kim; Sørensen, Trine Holst; Tokin, Radina ; Kari, Jeppe; Badino, Silke Flindt; Cavaleiro, Mafalda A. ; Røjel, Nanna Sandager; Christensen, Stefan Jarl; Vesterager, Cynthia Segura; Cola, Corinna Schiano di.

I: Biotechnology and Bioengineering (Print), Bind 115, Nr. 4, 01.2018, s. 831-838.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Thermoactivation of a cellobiohydrolase

AU - Westh, Peter

AU - Borch, Kim

AU - Sørensen, Trine Holst

AU - Tokin, Radina

AU - Kari, Jeppe

AU - Badino, Silke Flindt

AU - Cavaleiro, Mafalda A.

AU - Røjel, Nanna Sandager

AU - Christensen, Stefan Jarl

AU - Vesterager, Cynthia Segura

AU - Cola, Corinna Schiano di

N1 - This article has been found as a ’Free Version’ from the Publisher on January 4th 2019. When access to the article closes, please notify rucforsk@ruc.dk

PY - 2018/1

Y1 - 2018/1

N2 - We have measured activity and substrate affinity of the thermostable cellobiohydrolase, Cel7A, from Rasamsonia emersonii over a broad range of temperatures. For the wild type enzyme, which does not have a Carbohydrate Binding Module (CBM), higher temperature only led to moderately increased activity against cellulose, and we ascribed this to a pronounced, temperature induced desorption of enzyme from the substrate surface. We also tested a "high affinity" variant of R. emersonii Cel7A with a linker and CBM from a related enzyme. At room temperature, the activity of the variant was similar to the wild type, but the variant was more accelerated by temperature and about two-fold faster around 70 °C. This better thermoactivation of the high-affinity variant could not be linked to differences in stability or the catalytic process, but coincided with less desorption as temperature increased. Based on these observations and earlier reports on moderate thermoactivation of cellulases, we suggest that better cellulolytic activity at industrially relevant temperatures may be attained by engineering improved substrate affinity into enzymes that already possess good thermostability.

AB - We have measured activity and substrate affinity of the thermostable cellobiohydrolase, Cel7A, from Rasamsonia emersonii over a broad range of temperatures. For the wild type enzyme, which does not have a Carbohydrate Binding Module (CBM), higher temperature only led to moderately increased activity against cellulose, and we ascribed this to a pronounced, temperature induced desorption of enzyme from the substrate surface. We also tested a "high affinity" variant of R. emersonii Cel7A with a linker and CBM from a related enzyme. At room temperature, the activity of the variant was similar to the wild type, but the variant was more accelerated by temperature and about two-fold faster around 70 °C. This better thermoactivation of the high-affinity variant could not be linked to differences in stability or the catalytic process, but coincided with less desorption as temperature increased. Based on these observations and earlier reports on moderate thermoactivation of cellulases, we suggest that better cellulolytic activity at industrially relevant temperatures may be attained by engineering improved substrate affinity into enzymes that already possess good thermostability.

UR - https://onlinelibrary.wiley.com/doi/full/10.1002/bit.26513

U2 - 10.1002/bit.26513

DO - 10.1002/bit.26513

M3 - Journal article

VL - 115

SP - 831

EP - 838

JO - Biotechnology and Bioengineering (Print)

JF - Biotechnology and Bioengineering (Print)

SN - 0006-3592

IS - 4

ER -