Co-Production of Crystalline Cellulose and Biofuel from Low-Lignin Biomass

Co-Production of Crystalline Cellulose and Biofuel from Low-Lignin Biomass | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Co-Production of Crystalline Cellulose and Biofuel from Low-Lignin Biomass Julia Cunniffe, Nathalie Lavoine, Jack Wang, Vanessa Rondon Berrio, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7077301/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 24 Jan, 2026 Read the published version in BioEnergy Research → Version 1 posted 5 You are reading this latest preprint version Abstract Lignocellulosic biorefining has traditionally focused on either converting biomass into sugars for fuels or isolating solid cellulose for bioproducts. However, cost-effective strategies to maximize sugar yields while preserving crystalline cellulose remain underexplored. This study addresses that gap by optimizing cellulase enzymes to the co-production of fermentable sugars and crystalline cellulose. Laboratory-scale results also informed a techno-economic analysis (TEA) to evaluate the feasibility of industrial-scale implementation. To this end, we developed a selective hydrolysis process that targets hemicelluloses and amorphous cellulose, while retaining crystalline regions, using an optimized enzyme cocktail tested on three feedstocks: unbleached hardwood pulp, wild-type poplar, and clustered regularly interspaced short palindromic repeats (CRISPR)-edited poplar. Optimization across varying pH and temperature conditions enabled effective selective hydrolysis. Low-lignin unbleached pulp and CRISPR-edited poplar exhibited improved enzymatic accessibility and required less pretreatment, resulting in higher sugar yields and more efficient downstream processing. An engineered yeast strain co-fermented C5 and C6 sugars into ethanol, leaving behind high-crystallinity cellulose. CRISPR-edited poplar outperformed wild type, with 18% more sugar and 25% more ethanol yield, while enhancing cellulose crystallinity. TEA estimated crystalline cellulose production costs at $4,438 per metric tonne from unbleached pulp and $1,474 from CRISPR-edited biomass, highlighting the economic advantage of engineered feedstocks. This work presents a novel lignocellulosic biorefining approach that, for the first time, prioritizes the co-production of fermentable sugars and crystalline cellulose from low-lignin biomass. Low-lignin biomass Crystalline cellulose Biofuel Selective hydrolysis Fermentation Techno-economic analysis Full Text Supplementary Files CunniffeetalCoverLetter.docx Cite Share Download PDF Status: Published Journal Publication published 24 Jan, 2026 Read the published version in BioEnergy Research → Version 1 posted Editorial decision: Reconsider pending major revisions 30 Aug, 2025 Reviewers agreed at journal 20 Jul, 2025 Reviewers invited by journal 20 Jul, 2025 Editor assigned by journal 09 Jul, 2025 First submitted to journal 08 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7077301","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":488121367,"identity":"06f1d6ff-bd34-4342-af7a-cfb059388db7","order_by":0,"name":"Julia 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