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Title: Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure

Abstract

Glycoside hydrolase (GH) family 48 is an understudied and increasingly important exoglucanase family found in the majority of bacterial cellulase systems. Moreover, many thermophilic enzyme systems contain GH48 enzymes. Deletion of GH48 enzymes in these microorganisms results in drastic reduction in biomass deconstruction. Surprisingly, given their importance for these microorganisms, GH48s have intrinsically low cellulolytic activity but even in low ratios synergize greatly with GH9 endoglucanases. In this study, we explore the structural and enzymatic diversity of these enzymes across a wide range of temperature optima. We have crystallized one new GH48 module from Bacillus pumilus in a complex with cellobiose and cellohexaose (BpumGH48). We compare this structure to other known GH48 enzymes in an attempt to understand GH48 structure/function relationships and draw general rules correlating amino acid sequences and secondary structures to thermostability in this GH family.

Authors:
; ; ; ; ; ; ; ; ORCiD logo;
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1618693
Alternate Identifier(s):
OSTI ID: 1416714
Report Number(s):
NREL/JA-2700-70442
Journal ID: ISSN 1754-6834; 274; PII: 951
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Published Article
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Name: Biotechnology for Biofuels Journal Volume: 10 Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
Springer Science + Business Media
Country of Publication:
Netherlands
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; GH48; circular dichroism; x-ray crystallography; cellulase; molecular modeling

Citation Formats

Brunecky, Roman, Alahuhta, Markus, Sammond, Deanne W., Xu, Qi, Chen, Mo, Wilson, David B., Brady, John W., Himmel, Michael E., Bomble, Yannick J., and Lunin, Vladimir V. Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure. Netherlands: N. p., 2017. Web. doi:10.1186/s13068-017-0951-5.
Brunecky, Roman, Alahuhta, Markus, Sammond, Deanne W., Xu, Qi, Chen, Mo, Wilson, David B., Brady, John W., Himmel, Michael E., Bomble, Yannick J., & Lunin, Vladimir V. Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure. Netherlands. doi:10.1186/s13068-017-0951-5.
Brunecky, Roman, Alahuhta, Markus, Sammond, Deanne W., Xu, Qi, Chen, Mo, Wilson, David B., Brady, John W., Himmel, Michael E., Bomble, Yannick J., and Lunin, Vladimir V. Thu . "Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure". Netherlands. doi:10.1186/s13068-017-0951-5.
@article{osti_1618693,
title = {Natural diversity of glycoside hydrolase family 48 exoglucanases: insights from structure},
author = {Brunecky, Roman and Alahuhta, Markus and Sammond, Deanne W. and Xu, Qi and Chen, Mo and Wilson, David B. and Brady, John W. and Himmel, Michael E. and Bomble, Yannick J. and Lunin, Vladimir V.},
abstractNote = {Glycoside hydrolase (GH) family 48 is an understudied and increasingly important exoglucanase family found in the majority of bacterial cellulase systems. Moreover, many thermophilic enzyme systems contain GH48 enzymes. Deletion of GH48 enzymes in these microorganisms results in drastic reduction in biomass deconstruction. Surprisingly, given their importance for these microorganisms, GH48s have intrinsically low cellulolytic activity but even in low ratios synergize greatly with GH9 endoglucanases. In this study, we explore the structural and enzymatic diversity of these enzymes across a wide range of temperature optima. We have crystallized one new GH48 module from Bacillus pumilus in a complex with cellobiose and cellohexaose (BpumGH48). We compare this structure to other known GH48 enzymes in an attempt to understand GH48 structure/function relationships and draw general rules correlating amino acid sequences and secondary structures to thermostability in this GH family.},
doi = {10.1186/s13068-017-0951-5},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 10,
place = {Netherlands},
year = {2017},
month = {11}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1186/s13068-017-0951-5

Figures / Tables:

Fig. 1 Fig. 1: Circular Dichroism melt curves of three different GH48 catalytic domains. a C. bescii, b T. fusca, c B. pumilus

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    Works referencing / citing this record:

    Aromatic residues surrounding the active site tunnel of TfCel48A influence activity, processivity, and synergistic interactions with other cellulases
    journal, July 2019

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    Engineering Geobacillus thermoglucosidasius for direct utilisation of holocellulose from wheat straw
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      Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.