Redesigning plant cell walls for the biomass-based bioeconomy
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59 BASIC BIOLOGICAL SCIENCES↗
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Results for “saccharification”
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An Arabinoxylan Extracted from Corn Grain Is Inhibitory to Cellulase Activity
The maize seed expression system is a powerful platform for recombinant protein production. It has been utilized for high-yield production of industrial cellulases such as endo-1,4-b-Dglucanase (E1) and 1,4-b-D-glucan cellobiohydrolases (CBHI & CBHII). However, inhibitory compounds that could significantly reduce the cellulolytic efficiency were detected even in purified E1 and CBHII samples. After several steps of purification and analysis, the inhibitory compound found in the purified E1 sample had characteristics consistent with a thermostable polysaccharide with a molecular size over 10 kDa. Further polysaccharide analysis confirmed the molecular size of the detected polysaccharide and suggested that it is an arabinoxylan with a high arabinose/xylose ratio. Even though the strict column purification process should have removed water soluble non-protein molecules during protein purification, the inhibitor remained. Here, a possible explanation is that recombinant cellulase can bind to the inhibitor, carry it through all purification steps and finally release it when denatured by heat.
59 BASIC BIOLOGICAL SCIENCES↗
Conversion of poplar biomass into high-energy density tricyclic sesquiterpene jet fuel blendstocks
In an effort to ensure future energy security, reduce greenhouse gas emissions and create domestic jobs, the US has invested in technologies to develop sustainable biofuels and bioproducts from renewable carbon sources such as lignocellulosic biomass. Bio-derived jet fuel is of particular interest as aviation is less amenable to electrification compared to other modes of transportation and synthetic biology provides the ability to tailor fuel properties to enhance performance. Specific energy and energy density are important properties in determining the attractiveness of potential bio-derived jet fuels. For example, increased energy content can give the industry options such as longer range, higher load or reduced takeoff weight. Energy-dense sesquiterpenes have been identified as potential next-generation jet fuels that can be renewably produced from lignocellulosic biomass. We developed a biomass deconstruction and conversion process that enabled the production of two tricyclic sesquiterpenes, epi-isozizaene and prespatane, from the woody biomass poplar using the versatile basidiomycete Rhodosporidium toruloides. We demonstrated terpene production at both bench and bioreactor scales, with prespatane titers reaching 1173.6 mg/L when grown in poplar hydrolysate in a 2 L bioreactor. Additionally, we examined the theoretical fuel properties of prespatane and epi-isozizaene in their hydrogenated states as blending options for jet fuel, and compared them to aviation fuel, Jet A. Our findings indicate that prespatane and epi-isozizaene in their hydrogenated states would be attractive blending options in Jet A or other lower density renewable jet fuels as they would improve viscosity and increase their energy density. Saturated epi-isozizaene and saturated prespatane have energy densities that are 16.6 and 18.8% higher than Jet A, respectively. These results highlight the potential of R. toruloides as a production host for the sustainable and scalable production of bio-derived jet fuel blends, and this is the first report of prespatane as an alternative jet fuel.
09 BIOMASS FUELS↗
Feedstock/pretreatment screening for bioconversion of sugar and lignin streams via deacetylated disc-refining
Recent publications have shown the benefits of deacetylation disc-refining (DDR) as a pretreatment process to deconstruct biomass into sugars and lignin residues. Major advantages of DDR pretreatment over steam and dilute acid pretreatment are the removal of acetyl and lignin during deacetylation. DDR does not generate hydroxymethylfurfural (HMF) and furfural which are commonly produced from steam and dilute acid pretreatments. Acetate, lignin, HMF, and furfural are known inhibitors during enzymatic hydrolysis and fermentation. Another advantage of deacetylation is the production of lignin-rich black liquor, which can be upgraded to other bioproducts. Furthermore, due to the lack of sugar degradation during deacetylation, DDR has significantly less sugar loss than other pretreatment methods. Previous studies for DDR have primarily focused on corn stover, but lacked the investigative studies of other feedstocks. This study was designed to screen various DDR process conditions at pilot scale using three different feedstocks, including corn stover, poplar, and switchgrass. The impact of the pretreatment conditions was evaluated by testing hydrolysates for bioconversion to 2,3-butanediol. Pretreatment of biomass by DDR showed high-conversion-yields and 2,3-BDO fermentation production yields. Techno-economic analysis (TEA) of the pretreatment for biomass to sugar was also developed based on NREL’s Aspen Model. This study shows that the cellulose and hemicellulose in poplar was more recalcitrant than herbaceous feedstocks which ultimately drove up the sugar cost. Switchgrass was also more recalcitrant than corn stover but less than poplar.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Using Incremental Changes to Convert Lignocellulosic Feedstocks to Cellulosic Ethanol
One billion tons of biomass feedstocks have been identified for the production of renewable biofuels and biochemicals. This is one of the key carbon feedstocks to supply energy to the transportation sector for light duty, heavy duty and aviation fuels. Utilization of lignocellulosic feedstocks supports an improved energy security by reducing demand of petroleum imports, agricultural development, job creation, and reducing greenhouse gas emissions. To date, however, operational challenges have stymied the industrial production of large volumes of lignocellulosic-based fuels and chemicals. As a result, significant research investment has been led by the United States Department of Energy to understand and improve operational reliability at pioneer cellulosic biorefineries. In this perspective article lignocellulosic conversion technologies are described that have been adopted from the starch ethanol process. The developed process culminated in successful demonstration of 1,000-h integrated runs using several feedstocks, including switchgrass, energy sorghum, and two types of corn kernel fiber. This report highlights process development that solved several of the issues that plagued—and continue to plague—many in the cellulosic sugars space such as biomass feeding into equipment, high ash content, diversified co-product value, and others.
09 BIOMASS FUELS↗
Solid-State Nuclear Magnetic Resonance as a Tool to Probe the Impact of Mechanical Preprocessing on the Structure and Arrangement of Plant Cell Wall Polymers
Efficient separation of the plant cell wall polymers during lignocellulose processing has been historically challenging due to insolubility of the polymers and their propensity for recalcitrant reassembly. Methods, such as “lignin first” extraction techniques, have advanced efficient biomass use, but the molecular mechanisms for recalcitrance remain enigmatic. Here, we discuss how solid-state Nuclear Magnetic Resonance (NMR) approaches report on the 3D organization of cellulose, xylan, and lignin in the plant cell wall. Recent results illustrate that the organization of these polymers varies across biomass sources and sample preparation methods, with even minimal physical processing causing significant effects. These structural differences contribute to variable extraction efficiencies for bioproducts after downstream processing. We propose that solid-state NMR methods can be applied to follow biomass processing, providing an understanding of the polymer rearrangements that can lead to poor yields for the desired bioproducts. The utility of the technique is illustrated for mechanical processing using lab-scale vibratory ball milling of Sorghum bicolor .
59 BASIC BIOLOGICAL SCIENCES↗
Downregulation of barley ferulate 5-hydroxylase dramatically alters straw lignin structure without impact on mechanical properties
Barley is a major cereal crop for temperate climates, and a diploid genetic model for polyploid wheat. Cereal straw biomass is an attractive source of feedstock for green technologies but lignin, a key determinant of feedstock recalcitrance, complicates bio-conversion processes. However, manipulating lignin content to improve the conversion process could negatively affect agronomic traits. An alternative approach is to manipulate lignin composition which influences the physical and chemical properties of straw. This study validates the function of a barley ferulate 5-hydroxylase gene and demonstrates that its downregulation using the RNA-interference approach substantially impacts lignin composition. We identified five barley genes having putative ferulate 5-hydroxylase activity. Downregulation of HvF5H1 substantially reduced the lignin syringyl/guaiacyl (S/G) ratio in straw while the lignin content, straw mechanical properties, plant growth habit, and grain characteristics all remained unaffected. Metabolic profiling revealed significant changes in the abundance of 173 features in the HvF5H1-RNAi lines. The drastic changes in the lignin polymer of transgenic lines highlight the plasticity of barley lignification processes and the associated potential for manipulating and improving lignocellulosic biomass as a feedstock for green technologies. On the other hand, our results highlight some differences between the lignin biosynthetic pathway in barley, a temperate climate grass, and the warm climate grass, rice, and underscore potential diversity in the lignin biosynthetic pathways in grasses.
59 BASIC BIOLOGICAL SCIENCES↗