The binding, synergistic and structural characteristics of BsEXLX1 for loosening the main components of lignocellulose: Lignin, xylan, and cellulose

Wang, Qun; Liang, Chen; Lin, Hui; Yu, Daobing; Shen, Qi; Wan, Li; Zhao, Yanfang

doi:10.1016/j.enzmictec.2016.06.017

Cited by 6 publications

(4 citation statements)

References 22 publications

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“…Our results with E191K, with its fourfold increase in affinity to arabinoxylan and its complete loss of activity in the coleoptile wall assay (but not the filter paper assay), provides a striking example of the potential importance of protein navigation within the heterogeneous and complex cell wall. This mutant may also be interesting to test in binding and enzymatic deconstruction of lignocellulose for potential biofuel production , or as a probe for NMR‐based studies of the plant cell wall.…”

Section: Discussionmentioning

confidence: 99%

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

Hepler

Cosgrove

2019

FEBS Letters

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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 ~ 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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Section: Discussionmentioning

confidence: 99%

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

Hepler

Cosgrove

2019

FEBS Letters

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“…Bs EXLX1 is a member of the EXLX family of bacterial expansin and produced by Bacillus subtilis . The D2 domain is essential to the binding of Bs EXLX1 to cellulose, while mutating residue-125, 126, or 171 indicates their importance for binding to lignin and cellulose . Unlike the previous Escherichia coli system, we have expressed soluble recombinant expansins in the Pichia pastoris system, , which have not only markedly improved the yield of protein but also modified the thermostability of proteins by glycosylation.…”

Section: Introductionmentioning

confidence: 99%

“…Considering the binding ability of expansin, an understanding of the expansin adsorption will greatly contribute to the understanding of the action mechanism. Current research on expansin adsorption mainly concentrates on the adsorption site, adsorbed amount after equilibrium, and real-time adsorption behavior on cellulose. ,− For example, we recently found that cellobiose and xylose can improve the adsorption of Bs EXLX1 on cellulose at low concentrations but disrupt adsorption at high concentrations . However, the adsorption of expansin on lignin is unknown and remains one of the important unanswered questions in the field.…”

Section: Introductionmentioning

confidence: 99%

“…11 The D2 domain is essential to the binding of BsEXLX1 to cellulose, 12 while mutating residue-125, 126, or 171 indicates their importance for binding to lignin and cellulose. 13 Unlike the previous Escherichia coli system, 14 we have expressed soluble recombinant expansins in the Pichia pastoris system, 15,16 which have not only markedly improved the yield of protein but also modified the thermostability of proteins by glycosylation. BsEXLX1 has attracted the attention in the field of lignocellulose biorefinery because of its high binding ability and synergistic action with cellulase.…”

Section: ■ Introductionmentioning

confidence: 99%

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Real-Time QCM-D Monitoring of the Adsorption–Desorption of Expansin on Lignin

Cui

Duan

et al. 2020

Langmuir

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Expansin has nonhydrolytic disruptive activity and synergistically acts with cellulases to enhance the hydrolysis of cellulose. The adsorption–desorption of expansin on noncellulosic lignin can greatly affect the action of expansin on lignocellulose. In this study, three lignins with different sources (kraft lignin (KL), sodium lignin sulfonate (SLS), and enzymatic hydrolysis lignin (EHL)) were selected as the substrates. The real-time adsorption–desorption of Bacillus subtilis expansin (BsEXLX1) on lignins was monitored using quartz crystal microgravimetry with dissipation (QCM-D). The effects of temperature and Tween 80 on the adsorption–desorption behaviors were also investigated. The results show that BsEXLX1 exhibited high binding ability on lignin and achieved maximum adsorption of 283.2, 273.8, and 266.9 ng cm–2 at 25 °C on KL, SLS, and EHL, respectively. The maximum adsorption decreased to 148.2–192.8 ng cm–2 when the temperature increased from 25 to 45 °C. Moreover, Tween 80 competitively bound to lignin and significantly prevented expansin adsorption. After irreversible adsorption of Tween 80, the maximum adsorption of BsEXLX1 greatly decreased to 33.3, 37.2, and 10.3 ng cm–2 at 25 °C on KL, SLS, and EHL, respectively. Finally, a kinetic model was developed to analyze the adsorption–desorption process of BsEXLX1. BsEXLX1 has a higher adsorption rate constant (k A) and a lower desorption rate constant (k D) on KL than on SLS and EHL. The findings of this study provide useful insights into the adsorption–desorption of expansin on lignin.

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Bio‐Tribology and Bio‐Lubrication of Plant Cell Walls

Dolan

Yakubov

Stokes

2018

Annual Plant Reviews Online

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This article considers the underpinning role of interactive forces at the cell wall and cellulose–fibre interfaces in the physiological and biophysical functionality of the various non‐cellulosic components that are present within the plant cell walls. We specifically introduce the use of tribological science (friction and lubrication) to obtain new insights into relative movements occurring between cells and between cellulose fibrils within the cell walls, and the potential influence on friction forces by pectin, hemicelluloses, and expansins. This approach may provide a basis for describing physical phenomena which define the system's behaviour during growth, morphogenesis, and mechanical deformation of plants.

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The binding, synergistic and structural characteristics of BsEXLX1 for loosening the main components of lignocellulose: Lignin, xylan, and cellulose

Cited by 6 publications

References 22 publications

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

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

Real-Time QCM-D Monitoring of the Adsorption–Desorption of Expansin on Lignin

Bio‐Tribology and Bio‐Lubrication of Plant Cell Walls

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