Strategies to reduce end-product inhibition in family 48 glycoside hydrolases

Chen, Mo; Bu, Lintao; Alahuhta, Markus; Brunecky, Roman; Xu, Qi; Lunin, Vladimir V.; Brady, John W.; Crowley, Michael F.; Himmel, Michael E.; Bomble, Yannick J.

doi:10.1002/prot.24965

Strategies to reduce end-product inhibition in family 48 glycoside hydrolases

Journal Article · Sun Jan 31 23:00:00 EST 2016 · Proteins

DOI:https://doi.org/10.1002/prot.24965· OSTI ID:1240085

Chen, Mo ^[1]; Bu, Lintao ^[2]; Alahuhta, Markus ^[2]; Brunecky, Roman ^[2]; Xu, Qi ^[2]; Lunin, Vladimir V. ^[2]; Brady, John W. ^[1]; Crowley, Michael F. ^[2]; Himmel, Michael E. ^[2]; Bomble, Yannick J. ^[2]

Cornell Univ., Ithaca, NY (United States)
National Renewable Energy Lab. (NREL), Golden, CO (United States)

Family 48 cellobiohydrolases are some of the most abundant glycoside hydrolases in nature. They are able to degrade cellulosic biomass and therefore serve as good enzyme candidates for biofuel production. Family 48 cellulases hydrolyze cellulose chains via a processive mechanism, and produce end products composed primarily of cellobiose as well as other cellooligomers (dp ≤ 4). The challenge of utilizing cellulases in biofuel production lies in their extremely slow turnover rate. A factor contributing to the low enzyme activity is suggested to be product binding to enzyme and the resulting performance inhibition. In this study, we quantitatively evaluated the product inhibitory effect of four family 48 glycoside hydrolases using molecular dynamics simulations and product expulsion free-energy calculations. We also suggested a series of single mutants of the four family 48 glycoside hydrolases with theoretically reduced level of product inhibition. As a result, the theoretical calculations provide a guide for future experimental studies designed to produce mutant cellulases with enhanced activity.

View Accepted Manuscript (DOE)

Research Organization:: NREL (National Renewable Energy Laboratory (NREL), Golden, CO (United States))

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1240085

Report Number(s):: NREL/JA--2700-65427

Journal Information:: Proteins, Journal Name: Proteins Journal Issue: 3 Vol. 84; ISSN 0887-3585

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure Brunecky, Roman; Alahuhta, Markus; Sammond, Deanne W. Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0951-5	journal	November 2017
Determination of the native features of the exoglucanase Cel48S from Clostridium thermocellum Liu, Ya-Jun; Liu, Shiyue; Dong, Sheng Biotechnology for Biofuels, Vol. 11, Issue 1 https://doi.org/10.1186/s13068-017-1009-4	journal	January 2018