Advancing continuous enzymatic hydrolysis for improved biomass saccharification
Abstract Background A deployable, continuous enzymatic hydrolysis (CEH) process can address cost and commercialization risks associated with second-generation (Gen2) biorefinery sugar/lignin/ethanol production while contributing to energy supply and security. Developments in commercial enzymatic hyd...
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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/s13068-025-02680-z |
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| author | Roman Brunecky Yudong Li Stephen R. Decker Michael E. Himmel |
| author_facet | Roman Brunecky Yudong Li Stephen R. Decker Michael E. Himmel |
| author_sort | Roman Brunecky |
| collection | DOAJ |
| description | Abstract Background A deployable, continuous enzymatic hydrolysis (CEH) process can address cost and commercialization risks associated with second-generation (Gen2) biorefinery sugar/lignin/ethanol production while contributing to energy supply and security. Developments in commercial enzymatic hydrolysis formulations targeting Gen2 pretreated biomass such as deacetylated mechanically refined (DMR) biomass necessitate a reassessment of the existing hybrid simultaneous saccharification and fermentation (SSF) approach. Notably, the practice of "finishing hydrolysis" in SSF has become problematic with the introduction of oxidative enzymes, such as lytic polysaccharide monooxygenases (LPMOs), into commercial cellulase formulations as these require specific redox conditions and cofactor. Moreover, continuous SSF has not been demonstrated at commercial scale, limiting deployment and the associated economic benefits to farmers, producers, and support industries. Results Continuous enzymatic hydrolysis (CEH) was demonstrated at bench scale to enable optimal saccharification performance of deacetylated mechanically refined (DMR) pretreated biomass. Diafiltration was demonstrated to retain pretreated biomass solids and enzymes for continuous reaction while removing solubilized product sugars in situ. A significant breakthrough afforded by the CEH process is its ability to achieve equivalent endpoint conversions with approximately 50% lower enzyme loading. Yields of glucose and xylose were increased ~ 15% and ~ 4%, respectively, over batch hydrolysis. Unlike SSF using yeast or Zymomonas, CEH allows precise optimization of pH, temperature, oxygen tension, LPMO mediator concentration, and removal of end-product inhibitors. Conclusions Advanced CEH holds promise as a transformational, process-intensified, and cost-effective method for producing soluble clarified biomass sugars and insoluble lignin-rich streams. Enhancing saccharification performance, optimizing operating parameters, and employing membrane filtration will help overcome existing challenges and enable the efficient production of valuable biomaterials from lignocellulosic biomass. |
| format | Article |
| id | doaj-art-902db3e6e5314ab185ff9dad1b3c821e |
| institution | Kabale University |
| issn | 2731-3654 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
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| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj-art-902db3e6e5314ab185ff9dad1b3c821e2025-08-20T03:46:07ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542025-07-0118111110.1186/s13068-025-02680-zAdvancing continuous enzymatic hydrolysis for improved biomass saccharificationRoman Brunecky0Yudong Li1Stephen R. Decker2Michael E. Himmel3Biosciences Center, National Renewable Energy LaboratoryCatalytic Carbon Transformation and Scale-Up Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryBiosciences Center, National Renewable Energy LaboratoryAbstract Background A deployable, continuous enzymatic hydrolysis (CEH) process can address cost and commercialization risks associated with second-generation (Gen2) biorefinery sugar/lignin/ethanol production while contributing to energy supply and security. Developments in commercial enzymatic hydrolysis formulations targeting Gen2 pretreated biomass such as deacetylated mechanically refined (DMR) biomass necessitate a reassessment of the existing hybrid simultaneous saccharification and fermentation (SSF) approach. Notably, the practice of "finishing hydrolysis" in SSF has become problematic with the introduction of oxidative enzymes, such as lytic polysaccharide monooxygenases (LPMOs), into commercial cellulase formulations as these require specific redox conditions and cofactor. Moreover, continuous SSF has not been demonstrated at commercial scale, limiting deployment and the associated economic benefits to farmers, producers, and support industries. Results Continuous enzymatic hydrolysis (CEH) was demonstrated at bench scale to enable optimal saccharification performance of deacetylated mechanically refined (DMR) pretreated biomass. Diafiltration was demonstrated to retain pretreated biomass solids and enzymes for continuous reaction while removing solubilized product sugars in situ. A significant breakthrough afforded by the CEH process is its ability to achieve equivalent endpoint conversions with approximately 50% lower enzyme loading. Yields of glucose and xylose were increased ~ 15% and ~ 4%, respectively, over batch hydrolysis. Unlike SSF using yeast or Zymomonas, CEH allows precise optimization of pH, temperature, oxygen tension, LPMO mediator concentration, and removal of end-product inhibitors. Conclusions Advanced CEH holds promise as a transformational, process-intensified, and cost-effective method for producing soluble clarified biomass sugars and insoluble lignin-rich streams. Enhancing saccharification performance, optimizing operating parameters, and employing membrane filtration will help overcome existing challenges and enable the efficient production of valuable biomaterials from lignocellulosic biomass.https://doi.org/10.1186/s13068-025-02680-zBiomassCellulaseEnzyme hydrolysisSaccharificationBiomass sugarsLignocellulose |
| spellingShingle | Roman Brunecky Yudong Li Stephen R. Decker Michael E. Himmel Advancing continuous enzymatic hydrolysis for improved biomass saccharification Biotechnology for Biofuels and Bioproducts Biomass Cellulase Enzyme hydrolysis Saccharification Biomass sugars Lignocellulose |
| title | Advancing continuous enzymatic hydrolysis for improved biomass saccharification |
| title_full | Advancing continuous enzymatic hydrolysis for improved biomass saccharification |
| title_fullStr | Advancing continuous enzymatic hydrolysis for improved biomass saccharification |
| title_full_unstemmed | Advancing continuous enzymatic hydrolysis for improved biomass saccharification |
| title_short | Advancing continuous enzymatic hydrolysis for improved biomass saccharification |
| title_sort | advancing continuous enzymatic hydrolysis for improved biomass saccharification |
| topic | Biomass Cellulase Enzyme hydrolysis Saccharification Biomass sugars Lignocellulose |
| url | https://doi.org/10.1186/s13068-025-02680-z |
| work_keys_str_mv | AT romanbrunecky advancingcontinuousenzymatichydrolysisforimprovedbiomasssaccharification AT yudongli advancingcontinuousenzymatichydrolysisforimprovedbiomasssaccharification AT stephenrdecker advancingcontinuousenzymatichydrolysisforimprovedbiomasssaccharification AT michaelehimmel advancingcontinuousenzymatichydrolysisforimprovedbiomasssaccharification |