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At least 37 records · Page 2

Determining the Conformation of Supported Complexes Using an 17 O TEDOR-like NMR Experiment

Dynamic nuclear polarization surface-enhanced nuclear magnetic resonance (NMR) spectroscopy has enabled the determination of the three-dimensional configuration of surface sites, in particular supported metal complexes of relevance to single-site heterogeneous catalysis. These approaches have chiefly leveraged the application of NMR double-resonance experiments that either reveal the complex conformation via point-to-point intramolecular distances between spin-labeled atoms or the complex-surface orientation via distances between the spins and the surface plane. Either method typically requires expensive isotope labeling and each reports on different structural features. The application of an experiment that simultaneously reveals both types of distances with chemical resolution would be ideal. Here, in this article, we describe an 17 O{ 1 H} pseudo-3D correlation experiment that achieves this goal. Specifically, Si–O–Si and Si–O–M oxygens are well-resolved by 17 O NMR; therefore, distances can be simultaneously measured radially, between Si– 17 O–M and the 1 H’s of the ligands, and vertically to the Si– 17 O–Si linkages of the silica support. We demonstrate the experiment using supported yttrium and zirconium complexes. Good agreement is obtained when comparing the experimental results to theoretical predictions from density functional theory calculations, highlighting the reliability of this relatively simple experiment.

alkyls

Compartmental Control of UDP-Araf Supply: Golgi Lumen UDP-Arabinopyranose Mutase Depletes Plant Cell Wall Araf Linkages

L-Arabinofuranose (Araf) is a major constituent of plant cell wall polysaccharides. UDP-Araf is generated from UDP-L-arabinopyranose (UDP-Arap) by a UDP-Arap mutase (UAM) on the cytosolic face of the Golgi and then imported into the Golgi lumen for utilization by arabinosyltransferases (AraTs). Yet, a substantial fraction of UDP-Arap is synthesized in the Golgi lumen by the UDP-xylose 4-epimerase (UXE), creating a topological paradox where luminal UDP-Arap must be exported to the cytosol for conversion to UDP-Araf by UAM and then re-imported into the Golgi lumen. To test the functional significance of this compartmentation, we mislocalized Arabidopsis (Arabidopsis thaliana) UAM1 to the Golgi lumen by fusing it to the UXE1 transmembrane domain. The fusion protein was present in the Golgi, and protease-protection assays on microsomes supported its luminal orientation. In wild-type and uam1 plants, Golgi-targeted UAM1 reduced total cell wall Arabinose without consistent changes in other monosaccharides. Linkage analyses showed that this decrease reflects selective loss of Araf residues, whereas Arap remain largely unchanged, consistent with reduced UDP-Araf availability for luminal AraTs. Under salt stress, expression of Golgi-targeted UAM1 in uam1 caused root-growth inhibition and maturation-zone swelling that exogenous L-Ara failed to rescue. These results support a hypothesis that the strong thermodynamic bias of the UAM reaction toward UDP-Arap requires spatial separation of UDP-Arap production from UDP-Araf formation to sustain UDP-Araf flux into Golgi arabinosylation and avoid futile intraluminal back-conversion.

59 BASIC BIOLOGICAL SCIENCES

A multifunctional sesquiterpene synthase integrates with cytochrome P450s to reinforce the terpenoid defense network in maize

Terpenoids, the largest and most structurally diverse class of plant natural products, play essential roles in maize defense and ecological interactions. In this study, we identified and functionally characterized a sesquiterpenoid-based defense pathway in maize centered on α-santalenoic acid, a pathogen-inducible sesquiterpenoid antibiotic. Using a combination of metabolite-based genome-wide association studies (mGWAS), linkage mapping, and heterologous expression assays, we identified ZmTPS9 as a multiproduct terpene synthase that primarily produces α-santalene and β-bisabolene. Sequence analysis and site-directed mutagenesis revealed that threonine at position 413 is critical for enzyme activity, with its deletion resulting in a complete loss of enzyme activity. The sesquiterpene hydrocarbons produced by ZmTPS9 are further oxidized by three cytochrome P450 monooxygenases, ZmCYP71Z16, ZmCYP71Z18, and ZmCYP71Z19, to yield antimicrobial metabolites including α-santalenoic acid, zealexin D1 (ZD1), and zealexin D2 (ZD2). Together, these findings demonstrate a convergent biosynthetic strategy in maize, where multiproduct terpene synthases and promiscuous P450s collaboratively generate a flexible and robust terpenoid defense network.

a-santalenoic acid

Enzymatic Biocontrol of Fire Blight ( Erwinia amylovora ) Using an Engineered Glycosyl Hydrolase

Current management of fire blight, caused by Erwinia amylovora , relies heavily on streptomycin a practice that has contributed to the emergence of antibiotic-resistant strains and raised environmental and regulatory concerns. Enzyme-based biocontrol agents offer a promising antibiotic-free approach that combines target specificity with environmental compatibility. This study evaluates CAase, a bacteriophage-derived glycosyl hydrolase, for its ability to disrupt E. amylovora biofilms and reduce disease severity. Biochemical assays and scanning electron microscopy confirmed that CAase efficiently degraded the extracellular polysaccharide (EPS) matrix, releasing cells from biofilms. Gas chromatography–mass spectrometry (GC–MS) linkage analysis of EPS isolated from two Erwinia amylovora strains demonstrated that CAase preferentially cleaves galactan-rich amylovoran produced by strain EA273, while exhibiting only limited activity toward the levan-rich EPS from strain EA1430. Functional assays revealed that CAase reduced bacterial viability by nearly 2 orders of magnitude at higher enzyme concentrations, strongly suppressed surface motility, and induced ultrastructural damage visible by transmission electron microscopy. Importantly, field trials showed that CAase significantly lowered blossom and shoot blight incidence under orchard conditions. These results highlight CAase as a potent enzyme-based strategy for reducing the virulence of E. amylovora and demonstrate its potential as a sustainable alternative to antibiotics in fire blight management.

antibiotic resistance

Stable single-site organonickel catalyst preferentially hydrogenolyses branched polyolefin C–C bonds

Current methods of processing accumulated polyolefin waste typically require harsh conditions, precious metals or high metal loadings to achieve appreciable activities. Here, in this work, we examined supported, single-site organonickel catalysts for polyolefin upcycling. Chemisorption of Ni(COD) 2 (COD, 1,5-cyclooctadiene) onto Brønsted acidic sulfated alumina (AlS) yields a highly electrophilic Ni(I) precatalyst, AlS/Ni(COD) 2 , which is converted under H 2 to the active AlS/Ni II H catalyst. This single-site system exhibits unique hydrogenolysis selectivity that favours cleaving branched polyolefin C–C linkages, enabling the hydrogenolytic separation of polyethylene and isotactic polypropylene (iPP) mixtures. Moreover, AlS/Ni II H remains highly selective and active for hydrogenolysis of iPP admixed with polyvinyl chloride, and the spent catalyst can be repeatedly regenerated by AlEt3 treatment. Experimental mechanistic analysis and density functional theory modelling reveal a turnover-limiting C–C scission pathway featuring β-alkyl transfer and strong olefin binding. These results highlight the potential of nickel-based systems for the selective upcycling of complex plastic waste streams.

green chemistry

Hempseed cell wall polysaccharides are dominated by linear xylans and cellulose: Comprehensive structural profiling of ten cultivars of industrial hemp, Cannabis sativa L .

Hempseed is a rich source of dietary fiber; however, there has been limited research on the variability of carbohydrate composition in hempseed cell walls. The primary aim of this study was to conduct a comprehensive chemical and structural analysis of the cell wall polysaccharides in ten hempseed cultivars. Water-soluble polysaccharides (WSP) and water-insoluble residues (WIR) were isolated and subsequently analyzed for their monosaccharide composition using HPAEC-PAD, glycosyl linkage analysis using GC–MS, and structural characterization via NMR spectroscopy. All hempseed cultivars contained a high proportion of insoluble fibers and smaller amounts of soluble polysaccharides. Glucose and xylose were the most abundant components of the WIR fractions, while the WSP fractions contained abundant amounts of galactose, galacturonic acid, arabinose, rhamnose, and mannose. The results of linkage and spectroscopic analysis were consistent with the compositional analysis, identifying cellulose and acetylated linear xylans as primary components of WIR, and arabinogalactans, rhamnogalacturonans, heteromannans, xyloglucans, and arabinan as predominant in WSP. Altogether, the study revealed a comparable cell wall structure among the analyzed hemp seed varieties. The high fiber content of whole hempseed-based ingredients presents significant potential for food manufacturers seeking to develop products with enhanced dietary fiber content, offering both functional and nutritional benefits for consumers.

59 BASIC BIOLOGICAL SCIENCES

Catalytic Activation of Pyran Ethers in Self-Solvating Saccharides with Alkaline Earth Metals (Final Report)

Catalytic activation of cellulose ethers occurs via hydroxyl-stabilized cleavage of inter-monomer glycosidic bonds. Cooperativity between two hydroxyl groups lowers the ether cleavage transition state necessary to break apart long carbohydrate chains to initiate small molecule formation as fuel precursors. Metals existing with lignocellulosic materials including alkaline earth metals (Ca 2+ , Mg 2+ ) or alkali (Na + , K + ) also can catalyze ether scission cooperatively. By bonding with carbohydrate hydroxyl groups, a metal cation disrupt the hydrogen bonding network and free carbohydrate functional groups to react; a second metal cation stabilizes the carbohydrate ether transition state and enhances the rate of polymer scission. In this work, we expanded our initial understanding of metal-catalyzed glycoside ether scission. Within polysaccharides such as cellulose, two ether groups exist: one between monomers (glycosidic linkages) and one within a pyran ring. The hydroxyl-group hydrogen bonding network and bound metal ions form low energy binding states that interact with both ether oxygens simultaneously. Based on this interaction, our primary hypothesis was that the competition between stabilizing the two ether scission transition states determines the extent of the major pathways. Metals that disrupt the hydrogen bonding network of the C6 of sugars and stabilize the glycosidic ether promote transglycosylation to levoglucosan; in contrast, metals that stabilize the pyran ether oxygen promote sugar ring fragmentation to furans such as furfural. Varying characteristics of metal cation catalysts and configurations of bound polysaccharide chains dictate the relative rates of ether scission.

09 BIOMASS FUELS

Nitrogen–Rich Conjugated Macrocycles: Synthesis, Conductivity, and Application in Electrochemical CO 2 Capture

Here we report a series of nitrogen-rich conjugated macrocycles that mimic the structure and function of semiconducting 2D metal–organic and covalent organic frameworks while providing greater solution processability and surface tunability. Using a new tetraaminotriphenylene building block that is compatible with both coordination chemistry and dynamic covalent chemistry reactions, we have synthesized two distinct macrocyclic cores containing Ni–N and phenazine-based linkages, respectively. The fully conjugated macrocycle cores support strong interlayer stacking and accessible nanochannels. For the metal–organic macrocycles, good out-of-plane charge transport is preserved, with pressed pellet conductivities of 10 –3 S/cm for the nickel variants. Lastly, using electrochemically mediated CO2 capture as an example, we illustrate how colloidal phenazine-based organic macrocycles improve electrical contact and active site electrochemical accessibility relative to bulk covalent organic framework powders. Together, these results highlight how simple macrocycles can enable new synthetic directions as well as new applications by combining the properties of crystalline porous frameworks, the processability of nanomaterials, and the precision of molecular synthesis.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH

High-Energy Hybridized States Enable Long-Lived Hot Electrons in Cobaloxime-Silicon Nanocrystal System

Strong electronic coupling is achieved between the molecular catalyst cobaloxime ([Co]) and silicon nanocrystals (Si NCs) bridged by an ethylenepyridine group derived from vinylpyridine (vpy) covalently bound to the Si NC surface (Si-vpy-[Co]). The ethylenepyridine tether in Si-vpy-[Co] is key to dramatic changes to the system’s physical properties which are not observed in the corresponding formylpyridine (fpy) system (Si-fpy-[Co]) consistent with strong electronic coupling previously observed only in dark electrochemical systems. UV−vis absorption spectroscopy reveals new [Co]-centered electronic states in Si-vpy-[Co], and transient absorption spectroscopy finds a strong absorption feature appearing within 250 fs and persisting for at least 5 ns. Astoundingly, spectroelectrochemical measurements reveal that this absorption feature is consistent with both the singly reduced [Co] − and doubly reduced [Co] 2− complexes, leading to the conclusion that these long-lived charges are derived from high-energy “hot” electrons residing in [Co]-centered states. Detailed analysis using cyclic voltammetry, spectroelectrochemistry, electron paramagnetic resonance spectroscopy, and density functional theory (DFT) calculations provides insight into the unique electronic structure created in Si-vpy-[Co]. DFT reveals that the new electronic states arise from hybridization between deep Si NC band states and high-energy molecular orbitals of the ethylenepyridine tether and the [Co] catalyst and are facilitated by σ-bonding character at the ethylenepyridine linkage. This study demonstrates that strong electronic coupling achieved through precise molecular chemistry can change the paradigm of otherwise fixed energy levels in hybrid photoelectrochemical systems for artificial photosynthesis and related applications.

14 SOLAR ENERGY

Allylation and Thermosetting of Acetosolv Wheat Straw Lignin

Abstract The acetosolv extraction, allylation and subsequent cross‐linking of wheat straw lignin to thermoset biomaterials is herein described. The extraction temperature proved to be of great importance for the quality of the resulting lignin, with moderate temperature being key for preservation of β‐O‐4’ linkages. The allylation of the acetosolv lignin was carried out using three different synthetic strategies, resulting in selective installation of either benzylic or phenolic allyl ethers, or unselective allylation of various hydroxyl groups via etherification and carboxyallylation. The different allylation protocols employed either allyl alcohol, allyl chloride, or diallylcarbonate as allyl precursors, with the latter resulting in the highest degree of functionalization. Selected allylated acetosolv lignins were cross‐linked using a thiol‐ene approach and the lignin with the highest density of allyl groups was found to form a cross‐linked thermoset material with properties comparable to kraft lignin‐based analogues.

Chemistry

Manufacturing of Fabric Electrodes using a High-Throughput Screening Platform for Redox Flow Batteries

The objective of this project is to establish a new manufacturing methodology with machine learning- based high-throughput screening for the design and development of hierarchical structured, high-performance fabric electrodes for redox flow batteries (RFBs). The end goal of the project is to design and manufacture fabric electrodes for RFB applications that can provide 250 mA/cm2 current density operation for 100-cycles with 80% average energy efficiency. This was accomplished by first examining the structure-performance-property linkages of the electrodes provided by our partner, AvCarb. The electrodes’ microstructure was characterized by determining their pore size distribution, tortuosity, specific surface area, and porosity. The ohmic, charge transfer and mass transfer resistances were then calculated using electrochemical impedance spectroscopy. Carbon cloth electrodes showed the greatest resistance, which was dominated by charge transfer resistance, which we believe is related to the surface functionalization. Full cell cycling was used in order to determine the area specific resistance and energy efficiency of the cells. All of this experimental data and the results of the mathematical model (to increase the amount of inputs with parametric sweeping) were used to develop a machine learning-based model for the design of high-performance fabric electrodes. Using the results from the machine learning tool, optimized electrodes were fabricated by AvCarb. The ohmic, charge transfer and mass transfer resistances for these new electrodes were measured, and both performed better than any of the initial samples which had been provided by AvCarb.

25 ENERGY STORAGE

An RNA ligase partner for the prokaryotic protein-only RNase P: insights into the functional diversity of RNase P from genome mining

RNase P can use either an RNA- or a protein-based active site to catalyze 5'-maturation of transfer RNAs (tRNAs). This distinctive attribute in the biocatalytic repertoire raises questions about the underlying evolutionary driving forces, especially if each variant somehow affords a selective advantage under certain conditions. Upon mining all publicly available prokaryotic genomes and examining gene co-occurrence, we discovered that an RNA ligase with circularization activity was significantly overrepresented in genomes that contain the protein form of RNase P. This unexpected linkage inspires testable ideas to understand the bases for scenarios that might favor RNase P variants of different architectures/make-up.

HARP

Plant Cell Wall Polysaccharide O-Acetyltransferases

Plant cell walls are largely composed of polysaccharide polymers, including cellulose, hemicelluloses (xyloglucan, xylan, mannan, and mixed-linkage β-1,3/1,4-glucan), and pectins. Among these cell wall polysaccharides, xyloglucan, xylan, mannan, and pectins are often O-acetylated, and polysaccharide O-acetylation plays important roles in cell wall assembly and disease resistance. Genetic and biochemical analyses have implicated the involvement of three groups of proteins in plant cell wall polysaccharide O-acetylation: trichome birefringence-like (TBL)/domain of unknown function 231 (DUF231), reduced wall acetylation (RWA), and altered xyloglucan 9 (AXY9). Although the exact roles of RWAs and AXY9 are yet to be identified, members of the TBL/DUF231 family have been found to be O-acetyltransferases responsible for the O-acetylation of xyloglucan, xylan, mannan, and pectins. Here, we provide a comprehensive overview of the occurrence of O-acetylated cell wall polysaccharides, the biochemical properties, structural features, and evolution of cell wall polysaccharide O-acetyltransferases, and the potential biotechnological applications of manipulations of cell wall polysaccharide acetylation. Further in-depth studies of the biochemical mechanisms of cell wall polysaccharide O-acetylation will not only enrich our understanding of cell wall biology, but also have important implications in engineering plants with increased disease resistance and reduced recalcitrance for biofuel production.

Plant Sciences

PSInet: a new global water potential network

Abstract Given the pressing challenges posed by climate change, it is crucial to develop a deeper understanding of the impacts of escalating drought and heat stress on terrestrial ecosystems and the vital services they offer. Soil and plant water potential play a pivotal role in governing the dynamics of water within ecosystems and exert direct control over plant function and mortality risk during periods of ecological stress. However, existing observations of water potential suffer from significant limitations, including their sporadic and discontinuous nature, inconsistent representation of relevant spatio-temporal scales and numerous methodological challenges. These limitations hinder the comprehensive and synthetic research needed to enhance our conceptual understanding and predictive models of plant function and survival under limited moisture availability. In this article, we present PSInet (PSI—for the Greek letter Ψ used to denote water potential), a novel collaborative network of researchers and data, designed to bridge the current critical information gap in water potential data. The primary objectives of PSInet are as follows. (i) Establishing the first openly accessible global database for time series of plant and soil water potential measurements, while providing important linkages with other relevant observation networks. (ii) Fostering an inclusive and diverse collaborative environment for all scientists studying water potential in various stages of their careers. (iii) Standardizing methodologies, processing and interpretation of water potential data through the engagement of a global community of scientists, facilitated by the dissemination of standardized protocols, best practices and early career training opportunities. (iv) Facilitating the use of the PSInet database for synthesizing knowledge and addressing prominent gaps in our understanding of plants’ physiological responses to various environmental stressors. The PSInet initiative is integral to meeting the fundamental research challenge of discerning which plant species will thrive and which will be vulnerable in a world undergoing rapid warming and increasing aridification.

Forestry

Beyond Solanaceae: incorporation of feruloyltyramine and feruloyloctopamine into Cannabaceae lignins

The ferulic acid amides, feruloyltyramine and feruloyloctopamine, have been widely reported as integral constituents in the lignins in several species of Solanaceae in which they function as authentic lignin monomers. In the present study, we demonstrate that these ferulic acid amides are likewise incorporated into the lignins of species within Cannabaceae, including hemp (Cannabis sativa), hops (Humulus lupulus), and European nettle tree (Celtis australis). Structural analyses using derivatization followed by reductive cleavage (DFRC) and two-dimensional nuclear magnetic resonance (2D-NMR) spectroscopy revealed that these ferulic acid amides are incorporated via 4−O- and 8−O-ether linkages, as well as through 8−5′ linkages forming phenylcoumaran structures. Examination of a broad phylogenetic range of plant families demonstrated the absence of these ferulic acid amides from the lignins of all families studied except Solanaceae and Cannabaceae. Given the distant phylogenetic relationship between Solanaceae and Cannabaceae, the recruitment of these ferulic acid amides as lignin monomers in both lineages likely constitutes a case for convergent evolution at the level of lignin biosynthetic pathways. The significance of these ferulic acid amides lies in their unique role as the sole nitrogen-containing phenolic compounds known to participate in lignin formation.

Cannabaceae

QTL Mapping of Seed Fatty Acid Contents in Camelina sativa Under Heat Stress

Heat stress alters oil quality in oilseed crops, yet its genetic underpinnings in Camelina sativa remain unclear. This study investigated the genetic basis of heat-induced changes in seed fatty acids using a recombinant inbred line (RIL) population derived from a cross between two camelina varieties, Suneson and Pryzeth. Exposure to high temperature during reproductive growth led to increased proportions of saturated (C16:0, C18:0) and monounsaturated (C18:1) fatty acids, whereas polyunsaturated C18:3, total unsaturated fatty acids (UFA) and the PUFA/MUFA ratio were decreased, suggesting an inhibition of the C18:1 → C18:2 → C18:3 desaturation pathway. A high-density linkage map (4981 bins across 20 chromosomes) was built, and 25 QTLs for fatty acids were detected, with hotspots on chromosomes 1, 9, 12, 13, 16, and 20. A major QTL on chromosome 1 (~ 80 cM) explained the largest variance component for PUFA/MUFA under heat. Three desaturase genes (FAD2, FAD7, FAD8) were located within key QTL intervals, nominating them as candidates for modulating unsaturation under elevated temperature. These results provide a genetic basis for fine mapping and functional validation, supporting future molecular and breeding efforts to stabilize oil quality under warming conditions.

Camelina

Soil microbial ecology and microbiome-metabolite linkages improve understanding of ecosystem states along terrestrial-aquatic interfaces

Coastal soils are dynamic systems where unique microbial niches are shaped by the intensity and duration of flooding between the terrestrial and aquatic boundaries of the terrestrial-aquatic interface (TAI). We aimed to understand the soil microbial community (16S rRNA gene) along the TAIs of a freshwater versus estuarine region and how it relates to organic matter (OM, via Fourier Transform Ion Cyclotron Resonance Mass Spectrometry). We studied the TAI gradients along a transect from upland (forested), transition (stressed forest), to wetland at three sites in each of the Lake Erie (freshwater) and Chesapeake Bay (estuarine) regions. Microbial communities differed significantly by region, transect position, and site. Contrary to expectations, given their dynamic hydrologies, transitions represented midpoints in microbial richness and diversity. We identified a core microbiome conserved across all transect positions within a region, highlighting potential microbial functions most resilient to environmental change. Indicator taxa unique to each transect position defined specific niches shaped by soil biogeochemistry. Co-expression networks of feature-level β-nearest-taxon indices revealed positive relationships in bacterial and OM feature contributions to community assembly. Our study provides critical insights into microbial communities at the forefront of hydrological changes in coastal areas that connect the land to lakes and oceans and remain vulnerable to changing weather patterns.

coastal ecosystems

A Post-translational Histidine–Histidine Cross-Link Enhances Enzymatic Oxygen Reduction Activity with Greater pH Adaptability

Cross-linked protein residues exist as enzyme cofactors to enable or enhance catalytic activities. Despite their importance in nature, the chemical identity of the cross-links is limited to certain amino acid combinations, whose function and the formation mechanism remain insufficiently understood due to the difficulty in isolating native enzymes without the cross-links. Herein, we report the formation and characterization of both His-Tyr and His-His cross-links under oxidative enzymatic turnover conditions in L29H/F33Y/F43H Mb, a structural and functional model of heme-copper oxidase (HCO). The connectivity of the cross-link was characterized as Ne ε2 (His29)-Cd δ2 (His43) by mass spectrometry (LC-MS/MS) and nuclear magnetic resonance (NMR). Interestingly, formation of the cross-link significantly enhances the oxygen reduction activity of the enzyme at neutral or basic pH with higher product specificity. X-ray crystallography has identified a novel Tyr-His cross-link through a Tyr-O-His linkage. Our mechanistic studies indicate the involvement of high-valent heme-iron and the neighboring tyrosine in an oxidative self-processing pathway to generate the cross-link. In conclusion, this work serves as a new example while providing insights into the enzyme cross-link formation, allowing the design of artificial biocatalysts containing these novel cross-links with higher activity and pH adaptability.

Liu, Yiwei [Univ. of Texas, Austin, TX (United Sta