Engineering Enhanced Cellobiohydrolase Activity

Steve Decker; Brandon Knott; Petri Alahuhta; Jeffrey Linger; Vladimir Lunin; Venkataramanan Subramanian; Qi Xu; Todd VanderWall; Michael Crowley; Michael Himmel; Gregg Beckham; Larry Taylor II; Sarah Hobdey; Kara Podkaminer; Logan Schuster; Yogesh Chaudhari; Wiiliam Adney; John Baker; Antonella Amore

doi:10.1038/s41467-018-03501-8

Engineering Enhanced Cellobiohydrolase Activity

Steve Decker, Brandon Knott, Petri Alahuhta, Jeffrey Linger, Vladimir Lunin, Venkataramanan Subramanian, Qi Xu, Todd VanderWall, Michael Crowley, Michael Himmel, Gregg Beckham, Larry Taylor II, Sarah Hobdey, Kara Podkaminer, Logan Schuster, Yogesh Chaudhari, Wiiliam Adney, John Baker, Antonella Amore

Research output: Contribution to journal › Article › peer-review

70 Scopus Citations

Abstract

Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure-activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.

Original language	American English
Article number	1186
Number of pages	10
Journal	Nature Communications
Volume	9
Issue number	1
DOIs	https://doi.org/10.1038/s41467-018-03501-8 https://doi.org/10.1038/s41467-018-03501-8
State	Published - 1 Dec 2018

Bibliographical note

Publisher Copyright:
© 2018 The Author(s).

NREL Publication Number

NREL/JA-2700-70605

Keywords

hydrolases
polysaccharides
protein design
x-ray crystallography

Access to Document

https://www.nrel.gov/docs/fy18osti/70605.pdf

Cite this

Decker, S., Knott, B., Alahuhta, P., Linger, J., Lunin, V., Subramanian, V., Xu, Q., VanderWall, T., Crowley, M., Himmel, M., Beckham, G., Taylor II, L., Hobdey, S., Podkaminer, K., Schuster, L., Chaudhari, Y., Adney, W., Baker, J., & Amore, A. (2018). Engineering Enhanced Cellobiohydrolase Activity. Nature Communications, 9(1), Article 1186. https://doi.org/10.1038/s41467-018-03501-8, https://doi.org/10.1038/s41467-018-03501-8

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abstract = "Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure-activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.",

keywords = "hydrolases, polysaccharides, protein design, x-ray crystallography",

author = "Steve Decker and Brandon Knott and Petri Alahuhta and Jeffrey Linger and Vladimir Lunin and Venkataramanan Subramanian and Qi Xu and Todd VanderWall and Michael Crowley and Michael Himmel and Gregg Beckham and {Taylor II}, Larry and Sarah Hobdey and Kara Podkaminer and Logan Schuster and Yogesh Chaudhari and Wiiliam Adney and John Baker and Antonella Amore",

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doi = "10.1038/s41467-018-03501-8",

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Decker, S , Knott, B , Alahuhta, P , Linger, J, Lunin, V, Subramanian, V , Xu, Q , VanderWall, T , Crowley, M , Himmel, M , Beckham, G, Taylor II, L, Hobdey, S, Podkaminer, K, Schuster, L, Chaudhari, Y, Adney, W, Baker, J & Amore, A 2018, 'Engineering Enhanced Cellobiohydrolase Activity', Nature Communications, vol. 9, no. 1, 1186. https://doi.org/10.1038/s41467-018-03501-8, https://doi.org/10.1038/s41467-018-03501-8

TY - JOUR

T1 - Engineering Enhanced Cellobiohydrolase Activity

AU - Decker, Steve

AU - Knott, Brandon

AU - Alahuhta, Petri

AU - Linger, Jeffrey

AU - Lunin, Vladimir

AU - Subramanian, Venkataramanan

AU - Xu, Qi

AU - VanderWall, Todd

AU - Crowley, Michael

AU - Himmel, Michael

AU - Beckham, Gregg

AU - Taylor II, Larry

AU - Hobdey, Sarah

AU - Podkaminer, Kara

AU - Schuster, Logan

AU - Chaudhari, Yogesh

AU - Adney, Wiiliam

AU - Baker, John

AU - Amore, Antonella

PY - 2018/12/1

Y1 - 2018/12/1

N2 - Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure-activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.

AB - Glycoside Hydrolase Family 7 cellobiohydrolases (GH7 CBHs) catalyze cellulose depolymerization in cellulolytic eukaryotes, making them key discovery and engineering targets. However, there remains a lack of robust structure-activity relationships for these industrially important cellulases. Here, we compare CBHs from Trichoderma reesei (TrCel7A) and Penicillium funiculosum (PfCel7A), which exhibit a multi-modular architecture consisting of catalytic domain (CD), carbohydrate-binding module, and linker. We show that PfCel7A exhibits 60% greater performance on biomass than TrCel7A. To understand the contribution of each domain to this improvement, we measure enzymatic activity for a library of CBH chimeras with swapped subdomains, demonstrating that the enhancement is mainly caused by PfCel7A CD. We solve the crystal structure of PfCel7A CD and use this information to create a second library of TrCel7A CD mutants, identifying a TrCel7A double mutant with near-equivalent activity to wild-type PfCel7A. Overall, these results reveal CBH regions that enable targeted activity improvements.

KW - hydrolases

KW - polysaccharides

KW - protein design

KW - x-ray crystallography

UR - http://www.scopus.com/inward/record.url?scp=85044265772&partnerID=8YFLogxK

U2 - 10.1038/s41467-018-03501-8

DO - 10.1038/s41467-018-03501-8

M3 - Article

C2 - 29567941

AN - SCOPUS:85044265772

SN - 2041-1723

VL - 9

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 1186

ER -

Engineering Enhanced Cellobiohydrolase Activity

Abstract

Bibliographical note

NREL Publication Number

Keywords

Access to Document

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