Enzymatic approaches for diversifying bioproducts from cellulosic biomass.

Cellulosic biomass is the most abundantly available natural carbon-based renewable resource on Earth. Its widespread availability, combined with rising awareness, evolving policies, and changing regulations supporting sustainable practices, has propelled its role as a crucial renewable feedstock to meet the escalating demand for eco-friendly and renewable materials, chemicals, and fuels. Initially, biorefinery models using cellulosic biomass had focused on single-product platform, primarily monomeric sugars for biofuel. However, since the launch of the first pioneering cellulosic plants in 2014, these models have undergone significant revisions to adapt their biomass upgrading strategy. These changes aim to diversify the bioproduct portfolio and improve the revenue streams of cellulosic biomass biorefineries. Within this area of research and development, enzyme-based technologies can play a significant role by contributing to eco-design in producing and creating innovative bioproducts. This Feature Article highlights our strategies and recent progress in utilizing the biological diversity and inherent selectivity of enzymes to develop and continuously optimize sustainable enzyme-based technologies with distinct application approaches. We have advanced technologies for standalone platforms, which produce various forms of cellulose nanomaterials engineered with customized and enhanced properties and high yields. Additionally, we have tailored technologies for integration within a biorefinery concept. This biorefinery approach prioritizes designing tailored processes to establish bionanomaterials, such as cellulose and lignin nanoparticles, and bioactive molecules as part of a new multi-bioproduct platform for cellulosic biomass biorefineries. These innovations expand the range of bioproducts that can be produced from cellulosic biomass, transcending the conventional focus on monomeric sugars for biofuel production to include biomaterials biorefinery. This shift thereby contributes to strengthening the Bioeconomy strategy and supporting the achievement of several Sustainable Development Goals (SDGs) of the 2030 Agenda for Sustainable Development.

High yield biorefinery products from sugarcane bagasse: Prebiotic xylooligosaccharides, cellulosic ethanol, cellulose nanofibrils and lignin nanoparticles.

An integrated biorefining strategy was applied to fractionate Sugarcane bagasse (SCB) into its major constituents, enabling high-yield conversion of the fractionated materials into high-value coproducts alongside cellulosic ethanol. Pilot-scale steam explosion produced a hydrolysate rich in low molecular weight xylooligosaccharides that had a high in vitro efficacy as a prebiotic towards different bifidobacteria. Lignin recovered after alkaline treatment of the steam-exploded SCB was converted into uniform spherical lignin nanoparticles (11.3 nm in diameter) by a green mechanical method. The resulting cellulose was hydrolyzed at 17.5% (w/v) consistency and low enzyme loading (17.5 mg/g) to yield a pure glucose hydrolysate at a high concentration (100 g/L) and a cellulosic solid residue that was defibrillated by disc ultra-refining into homogeneous cellulose nanofibrils (20.5 nm in diameter). Statistical optimization of the cellulosic hydrolysate fermentation led to ethanol production of 67.1 g/L, with a conversion yield of 0.48 g/g and productivity of 1.40 g/L.h.

Co-production of xylo-oligosaccharides, xylose and cellulose nanofibrils from sugarcane bagasse.

This work investigated the integration of a two-stage hydrothermal treatment for production of xylooligosaccharides (XOS), a high-value product, into the isolation process of cellulose nanofibrils (CNF) from sugarcane bagasse. Under optimized conditions, the first stage yielded a XOS-rich, high purity hydrolysate and in the second stage only a xylose-rich hydrolysate could be obtained at high purity. The resulting solid cellulosic fraction was delignified and bleached to obtain a cellulose-rich pulp, which was mechanically defibrillated by disc ultra-refining to CNF. Except for the viscosity, the sugarcane CNF showed properties (i.e., thermal stability, crystallinity and diameter size) comparable or superior to the CNF prepared from commercial bleached eucalyptus Kraft pulp. In conclusion, the integration of the two-stage hydrothermal treatment is an efficient and promising strategy to obtain hemicellulose-derived high-value co-products in the process of isolating CNF. In addition, lignin was also recovered as a co-product with yield comparable to other biomass fractionation approaches.

Can we use short rotation coppice poplar for sugar based biorefinery feedstock? Bioconversion of 2-year-old poplar grown as short rotation coppice.

BACKGROUND: Feedstock cost is a substantial barrier to the commercialization of lignocellulosic biorefineries. Poplar grown using a short rotation coppice (SRC) system has the potential to provide a low-cost feedstock and economically viable sugar yields for fuels and chemicals production. In the coppice management regime, poplars are harvested after 2 years' growth to develop the root system and establish the trees. The biomass from these 2-year-old trees is very heterogeneous, and includes components of leaf, bark, branch, and wood chip. This material is quite different than the samples that have been used in most poplar bioconversion research, which come from mature trees of short rotation forestry (SRF) plantations. If the coppice management regime is to be used, it is important that feedstock growers maximize their revenue from this initial harvest, but the heterogeneous nature of the biomass may be challenging for bioconversion. This work evaluates bioconversion of 2-year-old poplar coppice and compares its performance to whitewood chips from 12-year-old poplar. RESULTS: The 2-year-old whole tree coppice (WTC) is comprised of 37% leaf, 9% bark, 12% branch, and 42% wood chip. As expected, the chemical compositions of each component were markedly different. The leaf has a low sugar content but is high in phenolics, ash, and extractives. By removing the leaves, the sugar content of the biomass increased significantly, while the phenolic, ash, and extractives contents decreased. Leaf removal improved monomeric sugar yield by 147 kg/tonne of biomass following steam pretreatment and enzymatic hydrolysis. Bioconversion of the no-leaf coppice (NLC) achieved a 67% overall sugar recovery, showing no significant difference to mature whitewood from forestry plantation (WWF, 71%). The overall sugar yield of NLC was 135 kg/tonne less than that of WWF, due to the low inherent sugar content in original biomass. An economic analysis shows the minimum ethanol selling price required to cover the operating cost of NLC bioconversion was $1.69/gallon. CONCLUSIONS: Leaf removal resulted in significant improvement in overall monomeric sugar production from SRC biomass. Leaf removal is essential to achieve good yields in bioconversion of poplar. Economic analysis suggests the NLC could be a reasonable feedstock provided it can be obtained at a discounted price.