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At least 55 records · Page 3
All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

Final Technical Report

The Department of Energy is interested in technologies that support the sustainable production of fuels, chemicals, and other bioproducts from plant biomass, to offset the nation’s reliance on fossil resources. The plant cell wall of energy crops provides the largest reservoir of raw materials for bioproducts. However, the widespread use of plant cell walls is hampered by their complexity and resistance to breakdown. To improve the productivity and cost-effectiveness of using energy crops to generate bioproducts, the fundamental problem of deconstructing plant cell walls must be addressed. This project developed and evaluated an innovative genetic modification technology to produce strategically designed enzymes that specifically accumulate in the plant cell wall. The resulting enzyme-engineered energy crops are expected to grow normally under natural conditions but break down more quickly and easily under high temperature during the production of biobased products. As such, this plant cell wall targeting enzyme engineering effort will reduce the cost of plant cell wall deconstruction and ultimately improve the economics of bioproducts. The overall objective of this project is to develop and evaluate the in-planta enzyme engineering technology to reduce lignocellulose deconstruction cost. The concept was first validated using tobacco plant, a model plant system that is typically used in lab testing for initial concept validation. Then the enzyme optimization was validated using switchgrass, the energy crop to be used to produce bioproducts. There are three specific objectives in this Phase I project: (1) validate the enzyme optimization concept using tobacco plant, a model plant system. (2) validate the enzyme optimization concept using switchgrass. (3) techno-economic analysis (TEA) for further scale-up application. By the end of this project, in-planta enzyme engineering was validated in both tobacco and switchgrass plants, with improved enzyme activity and saccharification efficiency. The in-planta enzyme engineering in Tabacco didn’t have a significant impact on plant growth and development. Transgenic tobacco plants with in-planta cellulose degrading enzymes showed higher biomass digestibility than wild type. Gene construction and transformation in switchgrass was much longer than expected, which delayed the research progress. Besides, in-planta engineering of lignin degrading enzyme is more challenging than cellulose degrading enzyme, in terms of expression detection. Expression of lignin degrading enzyme and cellulose degrading enzyme improved biomass yield and saccharification efficiency of switchgrass, respectively. It is promising to express both genes in switchgrass for optimized overall performance. According to the results of TEA, switchgrass biomass production cost is mainly attributed to by fertility and harvesting. Biomass production profit can increase up to 10-fold depending on biomass price. The PHA production profit is also sensitive to the biomass price. The proposed technology could potentially reduce the biomass deconstruction cost from 33% to 9% of PHA revenue, making the biomass-based PHA competitive to petroleum-based polymers even in case of relatively high biomass price of biomass. Therefore, cultivation of the genetically engineered self-deconstruction switchgrass for Polyhydroxyalkanoate (PHA) production could benefit switchgrass grower and PHA producer with attractive profits for both sectors. This new enzyme optimization approach will be beneficial for bioindustries that use energy crops as feedstocks. It will improve the economic viability of converting energy crops to renewable products that support a sustainable society and helps address the Nation’s long-term strategic needs for renewable products and reduction of reliance on fossil resources.

42 ENGINEERING↗
DOE OSTI2022

Engineering Curcumin Biosynthesis in Poplar Affects Lignification and Biomass Yield

Lignocellulosic biomass is recalcitrant toward deconstruction into simple sugars mainly due to the presence of lignin. By engineering plants to partially replace traditional lignin monomers with alternative ones, lignin degradability and extractability can be enhanced. Previously, the alternative monomer curcumin has been successfully produced and incorporated into lignified cell walls of Arabidopsis by the heterologous expression of DIKETIDE-CoA SYNTHASE (DCS) and CURCUMIN SYNTHASE2 (CURS2). The resulting transgenic plants did not suffer from yield penalties and had an increased saccharification yield after alkaline pretreatment. Here, we translated this strategy into the bio-energy crop poplar. Via the heterologous expression of DCS and CURS2 under the control of the secondary cell wall CELLULOSE SYNTHASE A8-B promoter (ProCesA8-B), curcumin was also produced and incorporated into the lignified cell walls of poplar. ProCesA8-B:DCS_CURS2 transgenic poplars, however, suffered from shoot-tip necrosis and yield penalties. Compared to that of the wild-type (WT), the wood of transgenic poplars had 21% less cellulose, 28% more matrix polysaccharides, 23% more lignin and a significantly altered lignin composition. More specifically, ProCesA8-B:DCS_CURS2 lignin had a reduced syringyl/guaiacyl unit (S/G) ratio, an increased frequency of p-hydroxyphenyl (H) units, a decreased frequency of p-hydroxybenzoates and a higher fraction of phenylcoumaran units. Without, or with alkaline or hot water pretreatment, the saccharification efficiency of the transgenic lines was equal to that of the WT. These differences in (growth) phenotype illustrate that translational research in crops is essential to assess the value of an engineering strategy for applications. Further fine-tuning of this research strategy (e.g., by using more specific promoters or by translating this strategy to other crops such as maize) might lead to transgenic bio-energy crops with cell walls more amenable to deconstruction without settling in yield.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2000

Ozonation and alkaline-peroxide pretreatment of wheat straw for Cryptococcus curvatus fermentation

Crop residues in an Advanced Life Support System (ALS) contain many valuable components that could be recovered and used. Wheat is 60% inedible, with approximately 90% of the total sugars in the residue cellulose and hemicellulose. To release these sugars requires pretreatment followed by enzymatic hydrolysis. Cryptococcus curvatus, an oleaginous yeast, uses the sugars in cellulose and hemicellulose for growth and production of storage triglycerides. In this investigation, alkaline-peroxide and ozonation pretreatment methods were compared for their efficiency to release glucose and xylose to be used in the cultivation of C. curvatus. Leaching the biomass with water at 65 degrees C for 4 h prior to pretreatment facilitated saccharification. Alkaline-peroxide and ozone pretreatment were almost 100% and 80% saccharification efficient, respectively. The sugars derived from the hydrolysis of alkaline-peroxide-treated wheat straw supported the growth of C. curvatus and the production of edible single-cell oil.

Non-NASA Center↗
DOE OSTI2022

Assessing the availability of two bamboo species for fermentable sugars by alkaline hydrogen peroxide pretreatment

We report this study comprehensively investigated two bamboo species (i.e. Neosinocalamus affinis and Phyllostachys edulis) in terms of their cell wall ultrastructure, chemical compositions, enzymatic saccharification, and lignin structure before and after alkaline hydrogen peroxide pretreatment (AHP). During AHP, Neosinocalamus affinis (NAB) had higher delignification than Phyllostachys edulis (PEB), and thus showed better enzymatic digestibility (93.05% vs 53.57% for glucan). The fundamental chemical behavior of the bamboo lignins was analyzed by fluorescence microscope (FM), confocal Raman microscope (CRM), molecular weight analysis, and 2D HSQC-NMR. Results indicated that the PEB has thicker cell wall and more concentrated lignin in its compound middle lamella and cell corner middle lamella than NAB. Moreover, PEB lignin contains more G units (S/G of 0.95), in evident contrast to that of NAB lignin (S/G of 1.30), which favor the formation of C–C linkages, thus impeding its degradation during the AHP.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Manipulation of Lignin Monomer Composition Combined with the Introduction of Monolignol Conjugate Biosynthesis Leads to Synergistic Changes in Lignin Structure

Abstract The complexity of lignin structure impedes efficient cell wall digestibility. Native lignin is composed of a mixture of three dominant monomers, coupled together through a variety of linkages. Work over the past few decades has demonstrated that lignin composition can be altered through a variety of mutational and transgenic approaches such that the polymer is derived almost entirely from a single monomer. In this study, we investigated changes to lignin structure and digestibility in Arabidopsis thaliana in near-single-monolignol transgenics and mutants and determined whether novel monolignol conjugates, produced by a FERULOYL-CoA MONOLIGNOL TRANSFERASE (FMT) or a p-COUMAROYL-CoA MONOLIGNOL TRANSFERASE (PMT), could be integrated into these novel polymers to further improve saccharification efficiency. Monolignol conjugates, including a new conjugate of interest, p-coumaryl p-coumarate, were successfully integrated into high-H, high-G and high-S lignins in A. thaliana. Regardless of lignin composition, FMT- and PMT-expressing plants produced monolignol ferulates and monolignol p-coumarates, respectively, and incorporated them into their lignin. Through the production and incorporation of monolignol conjugates into near-single-monolignol lignins, we demonstrated that substrate availability, rather than monolignol transferase substrate preference, is the most important determining factor in the production of monolignol conjugates, and lignin composition helps dictate cell wall digestibility.

2D-HSQC-NMR↗
DOE OSTI2023

Simultaneous suppression of lignin, tricin and wall‐bound phenolic biosynthesis via the expression of monolignol 4‐ O ‐methyltransferases in rice

Summary Grass lignocelluloses feature complex compositions and structures. In addition to the presence of conventional lignin units from monolignols, acylated monolignols and flavonoid tricin also incorporate into lignin polymer; moreover, hydroxycinnamates, particularly ferulate, cross‐link arabinoxylan chains with each other and/or with lignin polymers. These structural complexities make grass lignocellulosics difficult to optimize for effective agro‐industrial applications. In the present study, we assess the applications of two engineered monolignol 4‐ O ‐methyltransferases (MOMTs) in modifying rice lignocellulosic properties. Two MOMTs confer regiospecific para ‐methylation of monolignols but with different catalytic preferences. The expression of MOMTs in rice resulted in differential but drastic suppression of lignin deposition, showing more than 50% decrease in guaiacyl lignin and up to an 90% reduction in syringyl lignin in transgenic lines. Moreover, the levels of arabinoxylan‐bound ferulate were reduced by up to 50%, and the levels of tricin in lignin fraction were also substantially reduced. Concomitantly, up to 11 μmol/g of the methanol‐extractable 4‐ O ‐methylated ferulic acid and 5–7 μmol/g 4‐ O ‐methylated sinapic acid were accumulated in MOMT transgenic lines. Both MOMTs in vitro displayed discernible substrate promiscuity towards a range of phenolics in addition to the dominant substrate monolignols, which partially explains their broad effects on grass phenolic biosynthesis. The cell wall structural and compositional changes resulted in up to 30% increase in saccharification yield of the de‐starched rice straw biomass after diluted acid‐pretreatment. These results demonstrate an effective strategy to tailor complex grass cell walls to generate improved cellulosic feedstocks for the fermentable sugar‐based production of biofuel and bio‐chemicals.

4-O-methylated↗
DOE OSTI2025

Engineering 2‐Pyrone‐4,6‐Dicarboxylic Acid Production Reveals Metabolic Plasticity of Poplar

Woody biomass is a promising source of fermentable sugars for biofuels and bio-based chemicals, but its industrial use is limited by the costly biorefinery process. A viable strategy to reduce costs involves enhancing both biomass processability and the generation of high-value co-products. Here, we report the implementation of a synthetic metabolic pathway in Populus tremula × P. alba to produce 2-pyrone-4,6-dicarboxylic acid (PDC), a key building block for biodegradable plastics and high-performance materials. This artificial pathway—comprising microbial genes AroG, QsuB, PmdA, PmdB, and PmdC—enabled de novo PDC production in the stems of transgenic poplar. Pathway expression also induced substantial metabolic reprogramming and altered cell wall composition. These include the hyperaccumulation of simple phenolics like protocatechuic acid (PCA) and vanillic acid (VA), alongside reduced levels of p-hydroxybenzoic acid. A large portion of VA was ester-linked to cell wall lignin, while PCA was incorporated into the lignin backbone, forming novel benzodioxane units; concurrently, lignin in transgenic plants exhibited a drastic reduction in guaiacyl- and syringyl-units, with a notable increase in p-hydroxyphenyl-units. Hemicellulose content, particularly xylan, was also significantly increased. Moreover, expression of the PDC-pathway led to the formation of novel VA-derived suberin aromatics, enhancing suberization in bark and roots and improving salt stress tolerance. These changes led to improved saccharification efficiency, with up to 25% more glucose and 2.5 times xylose released from woody biomass. These results demonstrate the metabolic flexibility of poplar and highlight its potential for engineering cost-effective, stress-resilient bioenergy crops with enhanced biorefinery traits.

2-pyrone-4↗
DOE OSTI2022

Rerouting of the lignin biosynthetic pathway by inhibition of cytosolic shikimate recycling in transgenic hybrid aspen

Lignin is a phenolic polymer deposited in the plant cell wall and is mainly polymerized from three canonical monomers (monolignols), i.e., p-coumaryl, coniferyl, and sinapyl alcohols. After polymerization, these alcohols form different lignin substructures. In dicotyledons, monolignols are biosynthesized from phenylalanine, an aromatic amino acid. Shikimate acts at two positions in the route to the lignin building blocks. It is part of the shikimate pathway which provides the precursor for the biosynthesis of phenylalanine, and is involved in the transesterification of pcoumaroyl- CoA to p-coumaroyl shikimate, one of key steps in the biosynthesis of coniferyl and sinapyl alcohols. The shikimate residue in p-coumaroyl shikimate is released in later steps, and the resulting shikimate becomes available again for the biosynthesis of new p-coumaroyl shikimate molecules. In this study, we inhibited cytosolic shikimate recycling in transgenic hybrid aspen by accelerated phosphorylation of shikimate in the cytosol through expression of a bacterial shikimate kinase. Here, this expression elicited an increase in p-hydroxyphenyl units of lignin and, by contrast, a decrease in guaiacyl and syringyl units. Transgenic plants with high shikimate kinase activity produced a lignin content comparable to that in wild-type plants and had an increased processability via enzymatic saccharification. Although expression of many genes was altered in the transgenic plants, elevated shikimate kinase activity did not exert a significant effect on the expression of the majority of genes responsible for lignin biosynthesis. The present results indicate that cytosolic shikimate recycling is crucial to the monomeric composition of lignin rather than for lignin content.

shikimate pathway↗
DOE OSTI2021

A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield

Lignification of secondary cell walls is a major factor conferring recalcitrance of lignocellulosic biomass to deconstruction for fuels and chemicals. Genetic modification can reduce lignin content and enhance saccharification efficiency, but usually at the cost of moderate-to-severe growth penalties. We have developed a method, using a single DNA construct that uses CRISPR–Cas9 gene editing to knock-out expression of an endogenous gene of lignin monomer biosynthesis while at the same time expressing a modified version of the gene’s open reading frame that escapes cutting by the Cas9 system and complements the introduced mutation in a tissue-specific manner. Expressing the complementing open reading frame in vessels allows for the regeneration of Arabidopsis plants with reduced lignin, wild-type biomass yield, and up to fourfold enhancement of cell wall sugar yield per plant. The above phenotypes are seen in both homozygous and bi-allelic heterozygous T1 lines, and are stable over at least four generations. The method provides a rapid approach for generating reduced lignin trees or crops with one single transformation event, and, paired with a range of tissue-specific promoters, provides a general strategy for optimizing loss-of-function traits that are associated with growth penalties. This method should be applicable to any plant species in which transformation and gene editing are feasible and validated vessel-specific promoters are available.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Field evaluation of transgenic hybrid poplars with desirable wood properties and enhanced growth for biofuel production by bicistronic expression of PdGA20ox1 and PtrMYB3 in wood-forming tissue

Background: To create an ideotype woody bioenergy crop with desirable growth and biomass properties, we utilized the viral 2A-meidated bicistronic expression strategy to express both PtrMYB3 ( MYB46 ortholog of Populus trichocarpa , a master regulator of secondary wall biosynthesis) and PdGA20ox1 (a GA20-oxidase from Pinus densiflora that produces gibberellins) in wood-forming tissue (i.e., developing xylem). Results: Transgenic Arabidopsis plants expressing the gene construct DX15::PdGA20ox1-2A-PtrMYB3 showed a significant increase in both stem fresh weight (threefold) and secondary wall thickening (1.27-fold) relative to wild-type (WT) plants. Transgenic poplars harboring the same gene construct grown in a greenhouse for 60 days had a stem fresh weight up to 2.6-fold greater than that of WT plants. In a living modified organism (LMO) field test conducted for 3 months of active growing season, the stem height and diameter growth of the transgenic poplars were 1.7- and 1.6-fold higher than those of WT plants, respectively, with minimal adverse growth defects. Although no significant changes in secondary wall thickening of the stem tissue of the transgenic poplars were observed, cellulose content was increased up to 14.4 wt% compared to WT, resulting in improved saccharification efficiency of the transgenic poplars. Moreover, enhanced woody biomass production by the transgenic poplars was further validated by re-planting in the same LMO field for additional two growing seasons. Conclusions: Taken together, these results show considerably enhanced wood formation of our transgenic poplars, with improved wood quality for biofuel production.

09 BIOMASS FUELS↗
DOE OSTI2022

Evaluation of engineered low-lignin poplar for conversion into advanced bioproducts

Lignocellulosic resources are promising feedstocks for the manufacture of bio-based products and bioenergy. However, the inherent recalcitrance of biomass to conversion into simple sugars currently hinders the deployment of advanced bioproducts at large scale. Lignin is a primary contributor to biomass recalcitrance as it protects cell wall polysaccharides from degradation and can inhibit hydrolytic enzymes via non-productive adsorption. Several engineering strategies have been designed to reduce lignin or modify its monomeric composition. For example, expression of bacterial 3-dehydroshikimate dehydratase (QsuB) in poplar trees resulted in a reduction in lignin due to redirection of metabolic flux toward 3,4-dihydroxybenzoate at the expense of lignin. This reduction was accompanied with remarkable changes in the pools of aromatic compounds that accumulate in the biomass. The impact of these modifications on downstream biomass deconstruction and conversion into advanced bioproducts was evaluated in the current study. Using ionic liquid pretreatment followed by enzymatic saccharification, biomass from engineered trees released more glucose and xylose compared to wild-type control trees under optimum conditions. Fermentation of the resulting hydrolysates using Rhodosporidium toruloides strains engineered to produce α-bisabolene, epi-isozizaene, and fatty alcohols showed no negative impact on cell growth and yielded higher titers of bioproducts (as much as + 58%) in the case of QsuB transgenics trees. Our data show that low-recalcitrant poplar biomass obtained with the QsuB technology has the potential to improve the production of advanced bioproducts.

09 BIOMASS FUELS↗
DOE OSTI2025

Integrated Green Biorefinery for the Production of Anthocyanins, Fermentable Sugars, and High Pure Lignin from Miscanthus × giganteus

Miscanthus x giganteus (Mxg) is a promising perennial crop for producing natural colorants, renewable fuels, and bioproducts. However, natural recalcitrance and high pretreatment cost are major barriers to their complete conversion. In this study, a green processing method has been investigated for efficient recovery of natural pigments (anthocyanins), fermentable sugars, and pure lignin from Mxg genotypes using choline chloride-based natural deep eutectic solvents (NADES) systems. Interestingly, choline chloride: lactic acid (ChCl: LA) NADES-processed biomass resulted in 67.8 ± 2.1 μg g –1 of anthocyanins from dry biomass. A maximum of 87.4%–94.1% glucose yield was achieved after enzymatic saccharification. The effective extraction of lignin with high purity with higher β-aryl ether (β—O—4) bonds from advanced crops is crucial for lignin valorization. Notably, highly pure lignin (≈93.4% ± 1.4%) is achieved after low-temperature NADES pretreatment while retaining lignin's native structure. 31 P nuclear magnetic resonance demonstrated that total phenolics for ChCl: LA-lignin resulted in 1.20 mmol g –1 hydroxyls. The relative monolignol composition of syringyl (S), guaiacyl (G), and p-hydroxyphenyl (H) is 19.0, 65.7, and 14.3%, respectively, as evidenced by heteronuclear single quantum coherence analysis. This study provides a novel approach for obtaining high-purity lignin for catalytic depolymerization for oligomers and bifunctional monoaromatics production and leverages current cellulosic biorefinery technologies.

Raj, Tirath [University of Illinois at Urbana-Cham↗
DOE OSTI2025

An overview of switchgrass phenotypes variability across diverse populations and their implications for conversion to fuels

There have been substantial changes to the human lifestyle over the past two centuries, which are reflected in the amount of fuel we consume to power our day-to-day needs. The way we use these resources has indeed manifested in an overdependence on non-renewable energy sources, such as coal and petroleum, for generating electricity and powering our transportation needs. There is a pressing need to explore alternative ways of fueling our current lifestyle without impacting the environment. Biofuels have long been touted as a sustainable solution for use as drop-in fuels in aviation and maritime applications. Still, they have yet to establish themselves as a competitive commercial alternative, necessitating further research and development. Lignocellulosic biomass is an underutilized resource that is widely accessible for the commercial processing of renewable biofuels. Bioenergy crops, such as switchgrass (Panicum virgatum L.), which can be cultivated on marginal lands with minimal competition for agricultural land, are an ideal and promising candidate for bulk-scale biofuel synthesis. Over the past 30 years, significant progress has been made in breeding and genetically modifying these grasses to enhance their drought resilience and subsequent yields. However, discrepancies in biomass composition can lead to irregular feedstocks for downstream operations, which in turn affect overall production targets for biofuels. Here, this review examines the variability in switchgrass (P. virgatum L.) biomass phenotypes across diverse populations and plant components, and their implications for biofuel conversion. The study highlights significant variations in biomass yield, composition, and cell wall chemistry both between switchgrass genotypes and within individual cultivars. Key findings include differences in cellulose, hemicellulose, and lignin content between leaves and stems, which affect biomass digestibility and ethanol yield. The review also discusses the impact of lignin chemistry, particularly the syringyl/guaicyl (S/G) ratio, on the efficiency of biomass saccharification. Furthermore, it explores how these variations respond differently to various pretreatment techniques, affecting overall biofuel production. We conclude that understanding and quantifying this variability is crucial for optimizing switchgrass as a feedstock for commercial biofuel production, thereby potentially addressing the pressing need for sustainable energy sources in sectors such as aviation.

Kousika, Rohit [Univ. of Tennessee, Knoxville, TN ↗
DOE OSTI2026

Harnessing plant‐based platform for low‐cost cellulosic sugar recovery from bioenergy crops

The price of pure cellulase enzyme for the recovery of fermentable cellulosic sugars is one of the major challenges that limit the commercialization of second-generation biofuels and bio-based products. This work shows a means to greatly reduce the cost of cellulases. The abundant capacity of plants to synthesize and hyperaccumulate transgenic proteins in their plastids has been demonstrated for two gene constructs using either NPT II or Tet C to regulate the expression of bacterial cellulase Cel6A. Previously, the expression of Cel6A to 20–35% of total soluble leaf protein in tobacco was shown in two consecutive field trials. Moving forward, the key challenge was the extraction of active enzymes from the transgenic tobacco leaves hyperaccumulating bacterial cellulases in a cost-effective manner. The study showed that unpurified crude extracts of NPT II and Tet C transgenic tobacco leaves recovered ~23% and ~29% w/w cellulosic sugars from energycane bagasse, respectively. A supplementation of as low as 25% of purified commercial cellulase improved the glucose recovery by 2.9 times (~85% w/w) compared to 100% crude extract, which is comparable to the glucose recovery obtained by commercial cellulases, thereby suggesting a reduced requirement of commercial cellulases leading to a cost reduction of 75% for biorefineries. Assuming a stable 40% cellulase yield in total soluble protein under field conditions and multiple harvests (one to three) a year, the study estimates that the potential cost for saccharification of 1 t of lignocellulosic biomass can be reduced to 67–200 USD by using crude leaf extracts of transgenic tobacco.

biofuel↗
DOE OSTI2025

Genetically engineered poplar wood effectively enhances the efficiency of deep eutectic solvent-mediated one-pot processing

Although lignocellulosic biomass is a renewable resource with the potential to replace fossil-derived fuels and chemicals, its recalcitrance, largely due to lignin, limits its utilization. Recent advancements in genetic engineering have produced transgenic trees with reduced lignin content and/or modified lignin structure without compromising growth traits. Here, three engineered poplar varieties are evaluated as feedstocks using a biocompatible one-pot deep eutectic solvent-mediated process that integrates biomass fractionation and enzymatic saccharification within a single reactor, eliminating water washing and reconditioning. All transgenic poplars exhibit higher fermentable sugar yields than wild-type (WT) trees. Notably, QsuB poplar, incorporating 3,4-dihydroxybenzoate in lignin, achieves the highest glucose conversion yield of 91.3% (vs. 73.0% from WT). AT5 and MdCHS3 poplars, incorporating ferulate esters and naringenin, also demonstrate improved glucose yields (86.7 and 84.7%, respectively), confirming reduced biomass recalcitrance. Additionally, residual lignins are valorized via hydrogenolysis into phenolic compounds, with comparable alkylphenol production across all lines. These findings demonstrate that the transgenic poplar lines not only serve as superior feedstocks for sugar conversion but also provide a rich resource for phenolic compound production, enhancing the operational and economic viability of integrated biorefinery processes.

biomasses↗
DOE OSTI2021

High-Biomass Sorghums as a Feedstock for Renewable Fuels and Chemicals

High-biomass sorghums are intended for use in biorefineries that convert vegetative biomass into renewable fuels and chemicals. The majority of plant biomass consists of cell walls, a complex matrix of cellulose, hemicellulosic polysaccharides and lignin. In the biorefinery, the biomass is ground and then subjected to a thermo-chemical pretreatment (elevated temperature, high pressure, chemical catalysts) that disrupts the cell wall matrix and solubilizes some of the cell wall polymers, followed by enzymatic saccharification of the cellulose, which produces D-glucose. The glucose and sometimes also the xylose derived from the hydrolysis of the hemicellulosic polysaccharides, are subsequently converted to fuels or other useful chemicals by microbial biocatalysts. The genetic improvement of high-biomass sorghums has as its ultimate goals to maximize the yield of fermentable sugars on a per-hectare basis, to minimize the inputs of fertilizer, irrigation, fungicides and pesticides and to reduce the environmental footprint. Breeding strategies thus need to target biomass yield, biomass composition, maturity, pest and disease resistance and nutrient use efficiency. Furthermore, this chapter reviews the genetic basis of these traits and their potential application in breeding programs.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2024

Thermophilic biocatalysts for one-step conversion of citrus waste into lactic acid

Agri-food residues offer significant potential as a raw material for the production of L-lactic acid through microbial fermentation. Weizmannia coagulans, previously known as Bacillus coagulans, is a spore-forming, lactic acid-producing, gram-positive, with known probiotic and prebiotic properties. This study aimed to evaluate the feasibility of utilizing untreated citrus waste as a sustainable feedstock for the production of L-lactic acid in a one-step process, by using the strain W. coagulans MA-13. By employing a thermophilic enzymatic cocktail (Cellic CTec2) in conjunction with the hydrolytic capabilities of MA-13, biomass degradation was enhanced by up to 62%. Moreover, batch and fed-batch fermentation experiments demonstrated the complete fermentation of glucose into L-lactic acid, achieving a concentration of up to 44.8 g/L. These results point to MA-13 as a microbial cell factory for one-step production of L-lactic acid, by combining cost-effective saccharification with MA-13 fermentative performance, on agri-food wastes. Moreover, the potential of this approach for sustainable valorization of agricultural waste streams is successfully proven.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2024

Reductive catalytic fractionation of cotton stalks: catalytic strategy for tuning the selectivity of phenolic monomers

Pretreatment of lignocellulosic biomass is a primary step to delink lignin from the lignin-carbohydrate complex for bioethanol production and other value-added products. Presently, these processes yield lignin (technical) with significant structural changes compared to native lignin and make it difficult to valorize. Reductive catalytic fractionation (RCF) is an advanced pretreatment process to valorize native lignin to selective phenolic monomers before capitalizing on carbohydrates. Herein, cotton stalks (CS) were pretreated with ethanol: water mixture catalyzed by bimetallic, Ru-Ni/HY (RNY) and Ru-Fe/HY (RFY), and trimetallic, Ru-Fe-Ni/HY (RFNY), catalysts using the endogenously produced H 2 . The favorable catalytic properties of the RFNY catalyst, including high reducibility, acidity, metal dispersion, the synergistic effect of all the metals, and appropriate pore and particle size resulted in the highest catalytic activity. Maximum 92% delignification efficiency was achieved with 91% holocellulose retention. The extracted lignin after depolymerization in the same step produced a 19 wt.% yield of phenolic monomers. Increasing the duration of the reaction from 2 to 6 h increased the selectivity to saturated alkyl chain phenolics (1 to 16 wt.%) at the expense of hemicellulose (50 to 9%). The fate of carbohydrates left behind after the RCF (CS-RCF) process was analyzed through enzymatic saccharification and compared with raw CS and acid-pretreated CS. In conclusion, CS-RCF yielded maximum glucose yield (45 g/L) within an incubation period of 72 h.

09 BIOMASS FUELS↗