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At least 307 records · Page 17

Rapid monitoring of fermentations: a feasibility study on biological 2,3-butanediol production

2,3-butanediol (2,3-BDO) is an economically important platform chemical that can be produced by the fermentation of sugars using an engineered strain of Zymomonas mobilis . These fermentations require continuous monitoring and modification of fermentation conditions to maximize 2,3-BDO yields and minimize the production of the undesired coproducts glycerol and acetoin. Because of the time required for sampling and off-line chromatographic measurement of fermentation samples, the ability of fermentation scientists to modify fermentation conditions in a timely manner is limited. The goal of this study was to test if near-infrared spectroscopy (NIRS) along with multivariate statistics could reduce the time needed for this analysis and enable real-time monitoring and control of the fermentation. In this work we developed partial least squares (PLS) calibration models to predict the concentrations of glucose, xylose, 2,3-BDO, acetoin, and glycerol in fermentations via NIRS using two different spectrometers and two different spectroscopy modalities. We first evaluated the feasibility of rapid NIRS monitoring through experiments where we measured the signals from each analyte of interest and built NIRS-based PLS models using spectra from synthetic samples containing uncorrelated concentrations of these analytes. All analytes showed unique spectral signatures, and this initial modeling showed that all analytes could be detected simultaneously. We then began work with samples from laboratory fermentation experiments and tested the feasibility of regression model development across two spectral collection modalities (at-line and on-line) and two instruments: a laboratory-grade instrument and a low-cost instrument with a more limited spectral range. All modalities showed promise in the ability to monitor Z. mobilis fermentations of glucose and xylose to 2,3-BDO. The low-cost instrument displayed a lower signal-to-noise ratio than the laboratory-grade instrument, which led to comparatively lower performance overall, but still provided sufficient accuracy to monitor fermentation trends. While the ease of use of on-line monitoring systems was favored as compared to at-line systems due to the lack of sampling required and potential for automated process control, we observed some decrease in performance due to the additional complexity of the sample matrix. We have demonstrated that NIRS combined with multivariate analysis can be used for at-line and on-line monitoring of the concentrations of glucose, xylose, 2,3-BDO, acetoin, and glycerol during Z. mobilis fermentations. The decrease in signal-to-noise ratio when using a low-cost spectrometer led to greater prediction error than the laboratory-grade spectrometer for at-line monitoring. The on-line monitoring modality showed great promise for real time process control via NIRS.

09 BIOMASS FUELS↗

Phenotypical modifications of immune cells are enhanced by extracellular matrix

Immune cells not only constitute tumour microenvironment but they may even affect disease prognosis as a result of dual functional roles that they may play in tumour tissues. Two frequently used established immune cell lines (lymphocytic Jurkat and monocytic THP-1) were used to test whether microenvironmental factors, especially molecular components of extracellular matrix, can shape the phenotype of immune cells. Proliferation, morphological and phenotypical analyses were applied to compare behaviour of the immune cells, typically cultured as suspensions in culture medium, with their behaviour in collagen type I-based and Matrigel-based 3D cultures. Density of both immune cell types in routine suspension cultures affected their subsequent proliferation in extracellular matrices. THP-1 cells appeared to be more sensitive to their surrounding microenvironment as judged from extracellular matrix type-dependent changes in their cell doubling times and from slight increase in their diameters in both extracellular matrix-containing cell cultures. Moreover, even chemically uninduced monocytic THP-1 cells were present in a minor fraction as CD68 positive cell population in collagen type I matrix indicating their partial differentiation to macrophages. Observed modifications of immune cells by microenvironmental factors may have profound implications for their roles in healthy and pathological tissues.

60 APPLIED LIFE SCIENCES↗

Detailed Method for the Purification of Rhamnogalacturonan-I (RG-I) in Arabidopsis thaliana

The plant cell wall is a dynamic and complex extracellular matrix that not only provides structural integrity and determines cell shape but also mediates intercellular communication. Among its major components, pectins play essential roles in cell adhesion, wall porosity, hydration, and flexibility. Rhamnogalacturonan-I (RG-I), a structurally diverse pectic polysaccharide, remains one of the least understood components of the plant cell wall. Its backbone is substituted with arabinan, galactan, and arabinogalactan side chains that vary in length, branching, and composition across tissues, species, and developmental stages. In addition, RG-I can undergo modifications such as backbone acetylation, further contributing to its structural complexity and functional diversity. To advance understanding of RG-I, we present a detailed method for isolating RG-I from the model plant Arabidopsis thaliana . Leveraging Arabidopsis as a model system provides major advantages owing to its well-characterized genome and powerful molecular toolkit, enabling deeper investigation into the roles of RG-I in plant development and responses to environmental stress. Our method consists of two major steps: an initial chemical extraction using oxalate, followed by endo-polygalacturonase (EPG) digestion to fragment the pectic domains. An advantage of this approach is that it produces a dry material that can be stored at room temperature without special handling and does not introduce chemicals that may interfere with downstream analyses. The purified RG-I can be used for detailed compositional and structural analyses, as well as for functional studies of enzymes involved in pectin biosynthesis, modification, and degradation. Although this protocol was developed for isolating RG-I from Arabidopsis rosette leaves, it is also applicable to other Arabidopsis organs and other plant species.

25 ENERGY STORAGE↗

Experimental and kinetic modeling study of tetralin: A naphtheno-aromatic fuel for gasoline, jet and diesel surrogates

Distillate fuels contain significant proportions of naphtheno-aromatic components and tetralin is a suitable surrogate component to represent this molecular moiety. The presence of aromatic and naphthyl rings makes kinetic modeling of tetralin very challenging. Primary radicals formed during the oxidation of tetralin can be aryl, benzylic or paraffinic in nature. Using available information on reaction paths and rate constants of naphthenes and alkyl-aromatics, in this study a kinetic model of tetralin has been developed with emphasis on low-temperature chemistry and high-pressure conditions. Due to the lack of high-level quantum chemical calculations on reaction pathways of tetralin, analogous rates from ab-initio studies on benzylic and paraffinic radicals have been adopted here. Some modifications to the reaction rate rules are incorporated to account for the unique characteristics of tetralin's molecular structure. Important reaction channels have been identified using reaction path and brute force sensitivity analyses. In order to investigate the model performance at low temperatures, new experiments are carried out in a rapid compression machine on blends of tetralin and 3-methylpentane. Blending of low-reactivity tetralin with a high-reactivity alkane allowed the investigation of tetralin ignition at very low temperatures (665 – 856 K). The kinetic model developed in the current study is found to predict the current experiments and literature data adequately. The new model will aid in high-fidelity surrogate predictions at engine-relevant conditions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Surface Modification of Backsheets Using Coupling Agents for Roll-To-Roll Processed Thin-Film Solar Photovoltaic (PV) Module Packaging Application

For many flexible electronic and photonic devices, moisture stability is one of the most important factors that affects its short- and long-term performance. To maintain the performance, the device should be packaged in such a way that it hermetically blocks moisture from the device; however, in practice, it is rather difficult to achieve. The more practical solution is to impede the moisture ingress to the device. In optoelectronic devices that will be outdoors like solar cells, the interfacial adhesion strength between the encapsulant layer (adhesive) and a moisture barrier layer is also a critical parameter. This paper presents surface modifications of poly(ethylene terephthalate) (PET) carrier films, one of the layers in the trilayer barrier film that directly adheres to an encapsulant, using chemical, UV/ozone, and both treatments to improve adhesion with the thermoset encapsulant polymer material. Whereas previous studies also utilized treatment methods to increase the wettability characteristics, in this paper, we not only present the results of the adhesion strength upon various techniques to achieve good adhesion but also screen their behavior upon exposure to a damp–heat (60 °C, 90% RH) environment. Here, we found that the combined treatment method increases the adhesion by up to 12.1-fold and demonstrates up to a 200% increase in adhesion strength even upon our severe damp–heat environmental condition.

36 MATERIALS SCIENCE↗

Decarboxylation of the Catalytic Lysine Residue by the C5α-Methyl-Substituted Carbapenem NA-1-157 Leads to Potent Inhibition of the OXA-58 Carbapenemase

Antibiotic resistance in bacteria is a major global health concern. The wide spread of carbapenemases, bacterial enzymes that degrade the last-resort carbapenem antibiotics, is responsible for multidrug resistance in bacterial pathogens and has further significantly exacerbated this problem. Acinetobacter baumannii is one of the leading nosocomial pathogens due to the acquisition and wide dissemination of carbapenem-hydrolyzing class D β-lactamases, which have dramatically diminished available therapeutic options. Thus, new antibiotics that are active against multidrug-resistantA. baumannii and carbapenemase inhibitors are urgently needed. Here we report characterization of the interaction of the C5α-methyl-substituted carbapenem NA-1-157 with one of the clinically important class D carbapenemases, OXA-58. Antibiotic susceptibility testing shows that the compound is more potent than commercial carbapenems against OXA-58-producingA. baumannii, with a clinically sensitive MIC value of 1 μg/mL. Kinetic studies demonstrate that NA-1-157 is a very poor substrate of the enzyme due mainly to a significantly reduced deacylation rate. Mass spectrometry analysis shows that inhibition of OXA-58 by NA-1-157 proceeds through both the classical acyl-enzyme intermediate and a reversible covalent species. Time-resolved X-ray crystallographic studies reveal that upon acylation of the enzyme, the compound causes progressive decarboxylation of the catalytic lysine residue, thus severely impairing deacylation. Overall, this study demonstrates that the carbapenem NA-1-157 is highly resistant to degradation by the OXA-58 carbapenemase.

Acinetobactor↗

Interpreting inorganic compositional depth profiles to understand the rate-limiting step in vapor phase infiltration processes

Vapor phase infiltration (VPI) is a post-polymerization modification technique that infuses inorganics into polymers to create organic–inorganic hybrid materials with new properties. Much is yet to be understood about the chemical kinetics underlying the VPI process. The aim of this study is to create a greater understanding of the process kinetics that govern the infiltration of trimethyl aluminum (TMA) and TiCl 4 into PMMA to form inorganic-PMMA hybrid materials. To gain insight, this paper initially examines the predicted results for the spatiotemporal concentrations of inorganics computed from a recently posited reaction–diffusion model for VPI. This model provides insight on how the Damköhler number (reaction versus diffusion rates) and non-Fickian diffusional processes (hindering) that result from the material transforming from a polymer to a hybrid can affect the evolution of inorganic concentration depth profiles with time. Subsequently, experimental XPS depth profiles are collected for TMA and TiCl 4 infiltrated PMMA films at 90 °C and 135 °C. The functional behavior of these depth profiles at varying infiltration times are qualitatively compared to various computed predictions and conclusions are drawn about the mechanisms of each of these processes. TMA infiltration into PMMA appears to transition from a diffusion-limited process at low temperatures (90 °C) to a reaction-limited process at high temperatures (135 °C) for the film thicknesses investigated here (200 nm). While TMA appears to fully infiltrate these 200 nm PMMA films within a few hours, TiCl 4 infiltration into PMMA is considerably slower, with full saturation not occurring even after 2 days of precursor exposure. Infiltration at 90 °C is so slow that no clear conclusions about mechanism can be drawn; however, at 135 °C, the TiCl 4 infiltration into PMMA is clearly a reaction-limited process, with TiCl 4 permeating the entire thickness (at low concentrations) within only a few minutes, but inorganic loading continuously increasing in a uniform manner over a course of 2 days. Near-surface deviations from the uniform-loading expected for a reaction-limited process also suggest that diffusional hindering is high for TiCl 4 infiltration into PMMA. In conclusion, these results demonstrate a new, ex situ analysis approach for investigating the rate-limiting process mechanisms for vapor phase infiltration.

36 MATERIALS SCIENCE↗

Cavity molecular dynamics simulations of vibrational polariton-enhanced molecular nonlinear absorption

We report recent experiments have observed that the chemical and photophysical properties of molecules can be modified inside an optical Fabry– Pérot microcavity under collective vibrational strong coupling (VSC) conditions, and such modification is currently not well understood by theory. In an effort to understand the origin of such cavity-induced phenomena, some recent studies have focused on the effect of the cavity environment on the nonlinear optical response of the molecular subsystem. Here, we use a recently proposed protocol for classical cavity molecular dynamics simulations to numerically investigate the linear and the nonlinear response of liquid carbon dioxide under such VSC conditions following an optical pulse excitation. We find that applying a strong pulse of excitation to the lower hybrid light–matter state, i.e., the lower polariton (LP), can lead to an overall molecular nonlinear absorption that is enhanced by up to two orders of magnitude relative to the excitation outside the cavity. This polariton-enhanced multiphoton absorption also causes an ultrashort LP lifetime (0.2 ps) under strong illumination. Unlike usual polariton relaxation processes—whereby polaritonic energy transfers directly to the manifold of singly excited vibrational dark states—under the present mechanism, the LP transfers energy directly to the manifold of higher vibrationally excited dark states; these highly excited dark states subsequently relax to the manifold of singly excited states with a lifetime of tens of ps. Because the present mechanism is generic in nature, we expect these numerical predictions to be experimentally observed in different molecular systems and in cavities with different volumes.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Regulation of translation by ribosomal RNA pseudouridylation

Pseudouridine is enriched in ribosomal, spliceosomal, transfer, and messenger RNA and thus integral to the central dogma. The chemical basis for how pseudouridine affects the molecular apparatus such as ribosome, however, remains elusive owing to the lack of structures without this natural modification. Here, we studied the translation of a hypopseudouridylated ribosome initiated by the internal ribosome entry site (IRES) elements. We analyzed eight cryo–electron microscopy structures of the ribosome bound with the Taura syndrome virus IRES in multiple functional states. We found widespread loss of pseudouridine-mediated interactions through water and long-range base pairings. In the presence of the translocase, eukaryotic elongation factor 2, and guanosine 5'-triphosphate hydrolysis, the hypopseudouridylated ribosome favors a rare unconducive conformation for decoding that is partially recouped in the ribosome population that remains modified at the P-site uridine. The structural principles learned establish the link between functional defects and modification loss and are likely applicable to other pseudouridine-associated processes.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluation of High Level Waste Sludge Processing Behavior

The U.S. Department of Energy’s (DOE) Hanford Site has 177 underground storage tanks that contain wastes from past nuclear fuel reprocessing and waste-management operations. Over 20% of this waste is in the form of an insoluble sludge that will require slurry modification before its transfer to the Waste Treatment and Immobilization Plant (WTP). Specific WTP acceptance criteria for waste feed delivery describe the physical and chemical characteristics of the waste that must be met before the waste is transferred to the WTP. One challenging requirement relates to the undissolved solids (UDS) composition in a waste feed because the waste contains solid particles that settle, and their concentration and relative proportion can change during the transfer of the waste in individual batches. A key uncertainty is the ability to transfer and mix wastes with large variations in UDS concentrations and resulting settling rates. To address this uncertainty, a number of small scale mixing and settling tests have been conducted to determine the mobilization performance of variable chemistry simulants. Comparison of the size and density of the particulate for each simulant to that of southeast area Hanford sludge was made using metrics for particle mobilization, suspension, settling, and pipeline transfer where dependance on particle size and density may be different, including: 1. Settling velocity, 2. Critical shear stress for erosion, 3. Just-suspended impeller speed, and 4. Pipeline critical transport velocity. Existing high-level waste sludge data has shown the effect that increasing Al concentration has on resulting settled solids. This differential settling of particles in the sludge has the possibility of resulting in solids segregation during feed preparation and uneven particle distribution during pipeline transportation or mixer jet pump operations. Understanding the predictive capabilities of HLW solids settling and transport as well as potential remedies for addressing disparate sludge behaviors can help provide technical guidance during HLW flowsheet planning.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Fluoropolymer Composites from Partially Perfluoroalkylated Waste Polyethylene

Chemically modified plastics have emerged as practical solutions to plastic waste increases. Here, the inherent novelty of decorating polymer chains with chemical functionality results in distinct properties that expand the available application space. Nevertheless, developing designer materials for specific applications beyond compatibilization or mild property enhancement is difficult due to the synergistic effects of both the polar functionality imparted and the parent materials' intrinsic properties. By incorporating perfluoro-alkyl side-chains onto the backbone of dehydrogenated waste HDPE, unique surface properties intermediate between polytetrafluoroethylene (PTFE, the model fluoropolymer) and HDPE become apparent, while the overall material mechanical and thermal properties result in more LLDPE-like materials. This is demonstrated through moderate decreases in the surface free energy of the perfluoroalkylated polyolefin surface (increase in H 2 O contact angle of ~ 6°) and increased ordering under shear when blended with PTFE nanoparticles where the crossover point occurred at higher strains. Critically, perfluoroalkylated HDPE possesses improved rheological modification properties at elevated temperatures with PTFE nanoparticles, resulting in more thermally robust and stable composite materials.

fluoropolymer↗

Valley splittings in Si/SiGe quantum dots with a germanium spike in the silicon well

Silicon-germanium heterostructures have successfully hosted quantum dot qubits, but the intrinsic near-degeneracy of the two lowest valley states poses an obstacle to high-fidelity quantum computing. Here we present a modification to the Si/SiGe heterostructure by the inclusion of a spike in germanium concentration within the quantum well in order to increase the valley splitting. The heterostructure is grown by chemical vapor deposition and magnetospectroscopy is performed on gate-defined quantum dots to measure the excited state spectrum. We demonstrate a large and widely tunable valley splitting as a function of applied vertical electric field and lateral dot confinement. We further investigate the role of the germanium spike by means of tight-binding simulations in single-electron dots and show a robust doubling of the valley splitting when the spike is present, as compared to a standard (spike-free) heterostructure. This doubling effect is nearly independent of the electric field, germanium content of the spike, and spike location. This experimental evidence of a stable, tunable quantum dot, despite a drastic change to the heterostructure, provides a foundation for future heterostructure modifications.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Modeling Approach for the Aluminum-clad Dry Storage Pilot using HFIR Fuel

To confirm that the dry storage of aluminum-clad research reactor spent nuclear fuel (ASNF) will remain within the safety envelope after applied drying schemes and that the resulting evolution of the gas space composition, temperature, and pressure conditions are understood, a dry storage pilot project is being established. The pilot will incorporate an instrumented lid for discrete interval or for on-demand gas composition and temperature monitoring of two DOE Standard Canisters (DSCs) loaded with three High Flux Isotope Reactor (HFIR) inner cores per DSC. Each DSC would be subjected to a separate alternative candidate drying scheme. Canisters will undergo 1 to 5 years of monitoring, including internal temperature and gas sampling to track pressure and composition changes. This report outlines the approach for modeling the ASNF-in-canister behavior in terms of evolving gas space conditions for the ASNF dry storage pilot using HFIR fuel. The ASNF has an adherent surface oxyhydroxide layer comprised of boehmite/bayerite that generates hydrogen when subjected to irradiation. Three-dimensional multi-physics computational fluid dynamics simulations will be executed to compute the thermal field within the DSC and provide inputs to a chemical model employed to compute pressure buildup as hydrogen is generated in the system. Implemented in Cantera, the chemical model solves gas phase and aluminum oxyhydroxide surface-mediated radiolysis reactions. Gas phase reactions are sourced from Wittman and Hanson (2015), whereas surface-mediated reactions are incorporated by fitting experimental data using an optimization algorithm (Abboud, 2023). Water radiolysis reactions from Wren and Ball (2001) are adopted with modifications as described in Abboud (2023c). Understanding the effect of the hydrogen buildup over time is important for long-term storage safety considerations. Modeling results will include the canister pressure, temperature, and composition evolution from the initial helium backfill with the addition of radiolytically-evolved chemical species (e.g., hydrogen and oxygen). The specific HFIR cores for the pilot program have not yet been selected, and the overall design is still in development. The CFD-chemical model used for this work will be based on prior models with necessary updates to allow for improved accuracy and efficiency. The experimental data obtained from the HFIR demonstration will be used to improve and validate the computational models to predict the ASNF-in-canister behavior.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Chemical processes for recovery and purification of high-purity uranium-234 from aged plutonium-238

This work describes a streamlined approach to the recovery and purification of 234 U from aged 238 Pu. Key modifications to conventional techniques are described that result in an efficient uranium and plutonium separation process that successfully produced tens of grams of ultrahigh purity 234 U at Oak Ridge National Laboratory. A new processing scheme was developed using an anion exchange column run in a cycling mode with HNO 3 media as the primary uranium purification process. This was followed with a uranyl peroxide precipitation step to purify and solidify the uranium before conversion to U 3 O 8 . Both the traditional uranyl peroxide precipitation method and a new cascade uranyl peroxide precipitation process were used for final purification of the 234 U.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

Phase-sensitive γ-encoded recoupling of heteronuclear dipolar interactions and 1 H chemical shift anisotropy

γ-encoded recoupling sequences are known to produce strong amplitude modulations that lead to sharp doublets when Fourier transformed. These doublets depend very little on the recoupled tensor asymmetry and thus enable for the straightforward determination of dynamic order parameters. It can, however, be difficult to measure small anisotropies, or small order parameters, using such sequences; the resonances from the doublet may overlap with each other, or with the zero-frequency glitch. This limitation has prevented the widespread use of 1 H chemical shift anisotropy (CSA) for the measurement of dynamics, particularly for CH protons which typically have CSAs of only a few ppm when immobile. In this work, we introduce a simple modification to the traditional 1 H CSA and proton-detected local field pulse sequences that enables the acquisition of a hypercomplex dataset and the removal of the uncorrelated magnetization that results in the zero-frequency glitch. These new sequences then yield a frequency shift in the indirect dimension, rather than a splitting, which is easily identifiable even in cases of weak interactions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Advanced Non-Tread Materials for Fuel-Efficient Tires

PPG Industries, Inc. proposes to develop a new silica filler that can increase fuel efficiency by 2% while maximizing key performance properties in non-tread tire components compared to current carbon black/silica filler blends. In this project PPG focused on a sidewall compound. The developed compounds/components will reduce energy losses by approximately 25%, with no more than a 5% loss of resistance to degradative forces (targeting better performance). PPG plans to achieve this goal by characterizing the impact the silica filler can have on rubber properties and then tuning the silica morphology and surface chemistry for this end use. PPG will also provide guidance in optimizing the compound formulation to make the best use of this new filler. A key aspect is to understand and maintain, if not improve, the aging performance as evaluated by crack growth resistance under ozone environment. Two key strategies were used to meet the rolling resistance goal while maintaining other performance properties. The first factor was understanding the surface treatments and chemical makeup of the silica and its effects on compound performance. The second factor was using the best performing silicas and determining the optimum compound modifications to achieve the final sidewall compound performance. Using statistical analysis throughout the optimization process, it was determined that changing the silica morphology played a role in improving the reduction of tear strength while reducing rolling resistance and the improvements observed varied depending on the CB type.

36 MATERIALS SCIENCE↗

Monolignol Benzoates Incorporate into the Lignin of Transgenic Populus trichocarpa Depleted in C3H and C4H

Lignin, an abundant renewable aromatic biopolymer, is an essential macromolecule in terrestrial vascular plants. Genetic modifications affecting monolignol biosynthesis, which produces lignin monomers, alter the metabolic flux through the pathway and are becoming increasingly explored for improving the lignin and biomass quality and for producing high-value commodity chemical feedstocks. Benzoate (BA) conjugates are important metabolites in plants that have not been adequately characterized as components of the lignin structure. In addition to finding trace levels of BA conjugates in wild-type (WT) Populus trichocarpa, these become significantly augmented in strategic triple transgenics. Genes for three key cytochrome P450 enzymes, two 4-hydroxylases (PtrC4H1 and PtrC4H2), and one 3-hydroxylase (PtrC3H3), were downregulated to produce the transgenic plants in which monolignol benzoate (ML-BA) conjugates were incorporated into their lignin at a level some 16-fold higher than in WT. The co-downregulation of PtrC4H1/PtrC4H2/PtrC3H3 genes may cause a rerouting or modification of the pathway due to the interaction of the benzoate biosynthetic pathway with the conventional pathway for monolignol biosynthesis. The total lignin content in the transgenics was decreased by 50%, and the lignin H-unit level was elevated some 70-fold. The wood from the transgenic trees showed reduced recalcitrance toward enzymatic saccharification, which has value for the conversion of plant biomass to bioenergy.

2D NMR↗