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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]
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  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); 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.
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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. https://doi.org/10.1002/1873-3468.13528
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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. https://doi.org/10.1002/1873-3468.13528. https://www.osti.gov/servlets/purl/1598355.
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@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 = {Thu Jul 04 00:00:00 EDT 2019},
month = {Thu Jul 04 00:00:00 EDT 2019}
}
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Journal Article:
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Publisher's Version of Record
https://doi.org/10.1002/1873-3468.13528
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Cited by: 5 works
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Figures / Tables:

Table 1 Table 1: Characterization of EXLX1 Variant Binding on Cellulose and Insoluble Arabinoxylan. Binding kinetics were calculated from three replicates. Mean ± SEM from fits of three independent binding assays (6 ≤ n ≤ 9). † binding below the detection limit.
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    Works referencing / citing this record:

    Expression of Two α-Type Expansins from Ammopiptanthus nanus in Arabidopsis thaliana Enhance Tolerance to Cold and Drought Stresses
    journal, October 2019

    • Liu, Yanping; Zhang, Li; Hao, Wenfang
    • International Journal of Molecular Sciences, Vol. 20, Issue 21
    • DOI: 10.3390/ijms20215255
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      Figures / Tables found in this record:

      Table 1 (p. 14)table
      Supplementary Table 1 (p. 15)figure
      Figure 1 (p. 19)figure
      Figure 2 (p. 20)figure
      Figure 3 (p. 21)figure
      Figure 4 (p. 22)figure
      Figure 5 (p. 23)figure
      Supplemental Figure 1 (p. 24)figure
      Supplemental Figure 2 (p. 25)figure
      Supplemental Figure 3 (p. 26)figure
      Supplemental Figure 4 (p. 27)figure
      Supplemental Figure 5 (p. 28)figure
      Supplemental Figure 6 (p. 29)figure
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