Journal
FEBS LETTERS
Volume 593, Issue 18, Pages 2545-2555Publisher
WILEY
DOI: 10.1002/1873-3468.13528
Keywords
bacterial expansin; cell wall loosening; cellulose binding; directed evolution; phage display
Funding
- US Department of Energy, Office of Science, Basic Energy Sciences [DE-FG02-84ER13179]
- Graduate Research Fellowship from the US National Science Foundation [DGE1255832]
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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 similar to 40% higher than wild-type. Mutant E191K exhibited stronger cellulose binding but failed to induce creep, evidently due to protein mistargeting. These results reveal the complexity of interactions between plant cell walls and wall-modifying proteins, an important consideration when engineering proteins for applications in biofuel production and plant pathogen resistance.
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