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

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.