Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls
Abstract Background Overcoming lignocellulose recalcitrance to enzymatic or chemical processing is a prerequisite for biorefinery applications. Expansins and loosenins are non-lytic proteins that could assist reducing this recalcitrance by disrupting the intermolecular contacts between plant cell wa...
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| Format: | Article |
| Language: | English |
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BMC
2025-07-01
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| Series: | Biotechnology for Biofuels and Bioproducts |
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| Online Access: | https://doi.org/10.1186/s13068-025-02674-x |
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| author | Pramod Sivan Deepika Dahiya Ylenia Jabalera Taru Koitto Raul Perez-Jimenez Ewa J. Mellerowicz Emma Master Francisco Vilaplana |
| author_facet | Pramod Sivan Deepika Dahiya Ylenia Jabalera Taru Koitto Raul Perez-Jimenez Ewa J. Mellerowicz Emma Master Francisco Vilaplana |
| author_sort | Pramod Sivan |
| collection | DOAJ |
| description | Abstract Background Overcoming lignocellulose recalcitrance to enzymatic or chemical processing is a prerequisite for biorefinery applications. Expansins and loosenins are non-lytic proteins that could assist reducing this recalcitrance by disrupting the intermolecular contacts between plant cell wall components. Here, immunolocalization with fluorescence and transmission electron microscopy (TEM) were used to study the ability of a Bacillus subtilis expansin-like protein (BsEXLX1), a Phanerochaete carnosa loosenin protein (PcaLOOL12) and a fusion protein of PcaLOOL12 with the carbohydrate-binding module 63 (CBM63) of BsEXLX1 (i.e., PcaLOOL12-CBM63) to bind secondary cell walls (SCW) of aspen fibres, including fresh aspen wood, milled wood fibres (MWF) and MWF subjected to subcritical water extraction. Results The immunofluorescence labelling of fresh wood samples showed a weak signal for PcaLOOL12 and a strong signal for BsEXLX1 and PcaLOOL12-CBM63, suggesting the importance of CBM63 for protein adsorption to SCW components. TEM analysis after immunogold labelling revealed the presence of BsEXLX1 and PcaLOOL12-CBM63 in all secondary cell wall layers. Pretreatment of wood samples with the proteins reduced the binding of glucomannan- and glucuronoxylan-specific monoclonal antibodies. Similarly, protein adsorption to MWF was higher before subcritical water extraction. Together, these results suggest the adsorption of BsEXLX1 and PcaLOOL12-CBM63 to SCWs was mediated at least in part by their interaction with hemicelluloses. Conclusions Our study demonstrates that microbial expansin-related proteins can bind to the secondary walls of aspen wood through potential interaction of CBM63 with hemicelluloses. |
| format | Article |
| id | doaj-art-77c2082d51cc4b2ba2e92cadc011c328 |
| institution | Kabale University |
| issn | 2731-3654 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj-art-77c2082d51cc4b2ba2e92cadc011c3282025-08-20T04:01:52ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542025-07-0118111010.1186/s13068-025-02674-xCarbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell wallsPramod Sivan0Deepika Dahiya1Ylenia Jabalera2Taru Koitto3Raul Perez-Jimenez4Ewa J. Mellerowicz5Emma Master6Francisco Vilaplana7Division of Glycoscience, Department of Chemistry, KTH Royal Institute of Technology, AlbaNova University CentreDepartment of Bioproducts and Biosystems, School of Chemical Engineering, Aalto UniversityCIC bioGUNE BRTADepartment of Bioproducts and Biosystems, School of Chemical Engineering, Aalto UniversityCIC bioGUNE BRTAUmeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural SciencesDepartment of Bioproducts and Biosystems, School of Chemical Engineering, Aalto UniversityDivision of Glycoscience, Department of Chemistry, KTH Royal Institute of Technology, AlbaNova University CentreAbstract Background Overcoming lignocellulose recalcitrance to enzymatic or chemical processing is a prerequisite for biorefinery applications. Expansins and loosenins are non-lytic proteins that could assist reducing this recalcitrance by disrupting the intermolecular contacts between plant cell wall components. Here, immunolocalization with fluorescence and transmission electron microscopy (TEM) were used to study the ability of a Bacillus subtilis expansin-like protein (BsEXLX1), a Phanerochaete carnosa loosenin protein (PcaLOOL12) and a fusion protein of PcaLOOL12 with the carbohydrate-binding module 63 (CBM63) of BsEXLX1 (i.e., PcaLOOL12-CBM63) to bind secondary cell walls (SCW) of aspen fibres, including fresh aspen wood, milled wood fibres (MWF) and MWF subjected to subcritical water extraction. Results The immunofluorescence labelling of fresh wood samples showed a weak signal for PcaLOOL12 and a strong signal for BsEXLX1 and PcaLOOL12-CBM63, suggesting the importance of CBM63 for protein adsorption to SCW components. TEM analysis after immunogold labelling revealed the presence of BsEXLX1 and PcaLOOL12-CBM63 in all secondary cell wall layers. Pretreatment of wood samples with the proteins reduced the binding of glucomannan- and glucuronoxylan-specific monoclonal antibodies. Similarly, protein adsorption to MWF was higher before subcritical water extraction. Together, these results suggest the adsorption of BsEXLX1 and PcaLOOL12-CBM63 to SCWs was mediated at least in part by their interaction with hemicelluloses. Conclusions Our study demonstrates that microbial expansin-related proteins can bind to the secondary walls of aspen wood through potential interaction of CBM63 with hemicelluloses.https://doi.org/10.1186/s13068-025-02674-xExpansinLooseninSecondary cell wallHemicelluloses |
| spellingShingle | Pramod Sivan Deepika Dahiya Ylenia Jabalera Taru Koitto Raul Perez-Jimenez Ewa J. Mellerowicz Emma Master Francisco Vilaplana Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls Biotechnology for Biofuels and Bioproducts Expansin Loosenin Secondary cell wall Hemicelluloses |
| title | Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls |
| title_full | Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls |
| title_fullStr | Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls |
| title_full_unstemmed | Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls |
| title_short | Carbohydrate-binding domain CBM63 of microbial expansin-like BsEXLX1 facilitates the adsorption of expansin-related proteins to hemicelluloses in plant secondary cell walls |
| title_sort | carbohydrate binding domain cbm63 of microbial expansin like bsexlx1 facilitates the adsorption of expansin related proteins to hemicelluloses in plant secondary cell walls |
| topic | Expansin Loosenin Secondary cell wall Hemicelluloses |
| url | https://doi.org/10.1186/s13068-025-02674-x |
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