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Title: Directed in vitro evolution of bacterial expansin BsEXLX1 for higher cellulose binding and its consequences for plant cell wall-loosening activities

Abstract

Expansins are cell wall-loosening proteins found in all land plants and many microbial species. Despite homologous structures, bacterial expansins have much weaker cellulose binding and wall-loosening activity than plant expansins. We hypothesized stronger cellulose binding would result in greater wall-loosening activity and used in vitro evolution of Bacillus subtilis BsEXLX1 to test this hypothesis. Mutants with stronger binding generally had greater wall-loosening activity, but the relationship was nonlinear and plateaued at ~ 40% higher than wild-type. Mutant E191K exhibited stronger cellulose binding but failed to induce creep, evidently due to protein mistargeting. These findings reveal the complexity of interactions between plant cell walls and wall-modifying proteins, a prime consideration when engineering proteins for applications in biofuel production and plant pathogen resistance.

Authors:
 [1];  [1]
  1. Pennsylvania State Univ., University Park, PA (United States)
Publication Date:
Research Org.:
Pennsylvania State Univ., University Park, PA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); National Science Foundation (NSF)
OSTI Identifier:
1598355
Alternate Identifier(s):
OSTI ID: 1543169
Grant/Contract Number:  
[FG02-84ER13179; DGE-1255832]
Resource Type:
Accepted Manuscript
Journal Name:
FEBS Letters
Additional Journal Information:
[ Journal Volume: 593; Journal Issue: 18]; Journal ID: ISSN 0014-5793
Publisher:
Federation of European Biochemical Societies
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; bacterial expansin; cell wall loosening; cellulose binding; directed evolution; phage display

Citation Formats

Hepler, Nathan K., and Cosgrove, Daniel J. Directed in vitro evolution of bacterial expansin BsEXLX1 for higher cellulose binding and its consequences for plant cell wall-loosening activities. United States: N. p., 2019. Web. doi:10.1002/1873-3468.13528.
Hepler, Nathan K., & Cosgrove, Daniel J. Directed in vitro evolution of bacterial expansin BsEXLX1 for higher cellulose binding and its consequences for plant cell wall-loosening activities. United States. doi:10.1002/1873-3468.13528.
Hepler, Nathan K., and Cosgrove, Daniel J. Thu . "Directed in vitro evolution of bacterial expansin BsEXLX1 for higher cellulose binding and its consequences for plant cell wall-loosening activities". United States. doi:10.1002/1873-3468.13528.
@article{osti_1598355,
title = {Directed in vitro evolution of bacterial expansin BsEXLX1 for higher cellulose binding and its consequences for plant cell wall-loosening activities},
author = {Hepler, Nathan K. and Cosgrove, Daniel J.},
abstractNote = {Expansins are cell wall-loosening proteins found in all land plants and many microbial species. Despite homologous structures, bacterial expansins have much weaker cellulose binding and wall-loosening activity than plant expansins. We hypothesized stronger cellulose binding would result in greater wall-loosening activity and used in vitro evolution of Bacillus subtilis BsEXLX1 to test this hypothesis. Mutants with stronger binding generally had greater wall-loosening activity, but the relationship was nonlinear and plateaued at ~ 40% higher than wild-type. Mutant E191K exhibited stronger cellulose binding but failed to induce creep, evidently due to protein mistargeting. These findings reveal the complexity of interactions between plant cell walls and wall-modifying proteins, a prime consideration when engineering proteins for applications in biofuel production and plant pathogen resistance.},
doi = {10.1002/1873-3468.13528},
journal = {FEBS Letters},
number = [18],
volume = [593],
place = {United States},
year = {2019},
month = {7}
}

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