Heteronuclear Correlation Research Articles

Refinement and complete solution NMR analysis of the structure of a 6-branched 1,3-β-D-glucan (OL-2) isolate from Omphalialapidescens

OL-2 is a water-soluble β-glucan produced by Omphalia lapidescens. This versatile glucan has potential applications in various industries, including food, cosmetics, and pharmaceuticals. In addition, OL-2 is known for its promising applications as a biomaterial and drug, owing to its reported antitumor and antiseptic properties. Although the biological activities of β-glucans vary depending on their primary structure, holistic clarification of OL-2 via solution NMR spectroscopy to ascertain its complete and unambiguous structure has not yet been achieved. In this study, a chain of solution NMR techniques, such as correlation spectroscopy, total correlation spectroscopy (TOCSY), nuclear Overhauser effect and exchange spectroscopy, 13C-edited heteronuclear single quantum coherence (HSQC), HSQC-TOCSY, heteronuclear multiple bond correlation, and heteronuclear 2-bond correlation pulse sequences were used to unambiguously assign all 1H and 13C atoms in OL-2. Based on our investigation, OL-2 consists of a 1,3-β-glucan backbone chain decorated with a single 6-branched β-glucosyl side unit on every fourth residue.

Profiling Enzyme Activity of l-Asparaginase II by NMR-Based Methyl Fingerprinting at Natural Abundance.

l-asparaginase II (MW 135 kDa) from E. coli is an FDA-approved protein drug used for the treatment of childhood leukemia. Despite its long history as a chemotherapeutic, the structural basis of enzyme action, in solution, remains widely contested. In this work, methyl-based 2D [1H-13C]-heteronuclear single-quantum correlation (HSQC) NMR, at natural abundance, has been used to profile the enzymatic activity of the commercially available enzyme drug. The [1H-13C]-HSQC NMR spectra of the protein reveal the role of a flexible loop segment in the activity of the enzyme, in solution. Addition of asparagine to the protein results in distinct conformational changes of the loop that could be signatures of intermediates formed in the catalytic reaction. To this end, an isothermal titration calorimetry (ITC)-based assay has been developed to measure the enzymatic reaction enthalpy, as a marker for its activity. Combining both ITC and NMR, it was shown that the disruption of the protein conformation can result in the loss of function. The scope, robustness, and validity of the loop fingerprints in relation to enzyme activity have been tested under different solution conditions. Overall, our results indicate that 2D NMR can be used reliably to gauge the structure-function of this enzyme, bypassing the need to label the protein. Such natural abundant NMR methods can be potentially extended to probe the structure-function aspects of high-molecular-weight protein therapeutics (glycosylated protein drugs, enzymes, therapeutic monoclonal antibodies, antibody-drug conjugates, and Fc-fusion proteins), where (a) flexible loops are required for their function and (b) isotope labeling may not be straightforward.

Optimal control derived sensitivity-enhanced CA-CO mixing sequences for MAS solid-state NMR – Applications in sequential protein backbone assignments

We have recently introduced optimal-control derived pulse sequences for sensitivity-enhanced heteronuclear correlation NMR experiments of solid proteins. Preservation of equivalent coherence transfer pathways using transverse-mixing pulses (TROP) in multidimensional pulse schemes allows to increase the sensitivity of the experiments by more than a factor of 2 per each indirect dimension. In this article, we present homonuclear CA-CO transverse-mixing elements (homoTROP) that are based on dipolar interactions and achieve similar gains as the heteronuclear TROP pulses described previously. Both transfer elements were subsequently implemented in 3D se-hCAcoNH and se-hCOcaNH, that together with the previously introduced 3D se-hCANH and se-hCONH experiments yield a complete set of sensitivity-enhanced protein backbone assignment experiments. In contrast to the J-coupling based methods that are used at fast (60 kHz) and ultrafast MAS (>100 kHz), the homoTROP experiments employ about 10-times shorter mixing times making use of the larger magnitude of the dipolar coupling in comparison to the J couplings. The experiments are demonstrated using a microcrystalline, perdeuterated sample of the chicken alpha-spectrin SH3 domain in which all exchangeable sites are fully back-substituted with protons. We evaluated the gains in efficiency in all experiments site-specifically observing that the se-hCAcoNH and se-hCOcaNH experiments yield an increase in sensitivity by a factor of 1.36±0.09 and at least a factor of 1.8 with respect to the conventional hcoCAcoNH and hCOcaNH J-based experiments.

Mapping Hydrogens in Hydrous Materials

Hydrous materials such as clays, cements, hydroxyapatite, and metal hydroxides have been known and extensively used since antiquity. However, experimental elucidation of hydrogen in hydrous materials remains challenging due to the intrinsic insensitivity, limited accessibility, sample damage, or poor spectral resolution of many characterization methods. 1H solid-state NMR spectroscopy has evolved into an ideal site-specific characterization tool of hydrogen-containing materials, but its current application in hydrous materials is hampered by the low resolution of 1H NMR spectra in the solid state due to the presence of intense dipolar coupling networks and a narrow 1H chemical shift range. Herein, a significant advance in the characterization of hydrogens in hydrous materials is reported. A combination of magic-angle spinning (MAS) NMR, moderate 2H substitution, and high magnetic fields has unlocked the capacity of identifying many chemically similar while crystallographically distinct hydrogen sites in a prototypical hydrous material Y4(OH)10Br2·3H2O (LYH-Br) by 1D 1H solid-state NMR experiments. 1H NMR peaks were first tentatively assigned in accordance with the density functional theory (DFT) calculation results, and the assignment was further refined by 2D 1H–89Y heteronuclear correlation (HETCOR) and 1H–1H double-quantum (DQ) MAS NMR data. The order of 1H chemical shift (δiso) values provides valuable information on the relative strength of hydrogen bond for ten hydroxide hydrogen sites. The power of very high spectral resolution is further described by observing all triple-quantum (TQ) coherences in the 2D 1H–1H TQ MAS NMR spectrum, in which all spatial proximities among three hydrogen sites are resolved. This demonstration of the very high spectral resolution obtained in 1D and 2D 1H solid-state NMR experiments illustrates how scientists in various communities can now study the multiple hydrous species of hydrous materials and obtain previously inaccessible fine structural information.

The study of structure and effects of two new proanthocyanidins from Anogeissus pendula leaves on rumen enzyme activities.

Two novel proanthocyanidins, (2R, 3R)-(+)-Gallocatechin-(4β → 8)4-(2R, 3R)-(+)-gallocatechin (compound 1) and 3-O-galloyl-(2S, 3S)-(-)-epicatechin-(4α → 8)-[3-O-galloyl-(2S, 3S)-(-)-epicatechin (4α → 8)]2-(2S, 3S)-(-)-epicatechin (compound 2), were structurally characterized from leaves of Anogeissus pendula. The structures were determined by ultraviolet spectroscopy (UV), proton nuclear magnetic resonance (1H NMR), 13C NMR, and heteronuclear multiple bond correlation. Molinspiration and Osiris property explorer applications were used to predict bioactivity and drug score. Drug scores of 0.08 and 0.05 were predicted for compounds 1 and 2, respectively. Predicted bioactivity scores were high. Due to their molecular weight, chemical structure, and conformation, the newly discovered proanthocyanidins possess an inclination to interact with proteins. Based on this premise, both compounds were subjected to in vitro testing against ruminal enzymes. They exhibited significant inhibition activities (p < 0.01) with a range of half maximal effective concentration (EC50) of 14.80-17.88 mg/mL of glutamic oxaloacetic transaminase in both protozoa and bacteria fractions. The ruminal glutamic pyruvic transaminase activity was significantly inhibited (p < 0.01) from EC50 12.59-16.29 mg/mL, and R-cellulase inhibition was recorded with EC50 18.20-21.98 mg/mL by compounds 1 and 2, respectively. Protease activity decreased with increasing incubation time and concentration of both compounds. The novel proanthocyanidins have potential roles in improving feed conversion ratios and in drug development.

Boron Adsorption Kinetics of Microcrystalline Cellulose and Polymer Resin

Tailoring boron-polysaccharide interactions is an important strategy for developing functional soft materials such as hydrogels, fire retardants, and sorbents for environmental remediation, for example, using lignocellulosic biomass. For such applications to be realized, it is paramount to understand the adsorption kinetics of borate anions on cellulose and their local structures. Here, the kinetic aspects of boron adsorption by microcrystalline cellulose, lignin, and polymeric resin are investigated and compared. Borate anions interact with the vicinal diols in the glucopyranoside moieties of cellulose to yield chemisorbed boron chelate complexes. In contrast to cellulose, technical lignin contains fewer cis-vicinal diols, and it does not have a tendency to form such chelate complexes upon treatment with the aqueous boric acid solution. The formation kinetics and stability of these chelate complexes strongly depend on nanoscale structures, as well as reaction conditions such as pH and concentration of the sorbate and sorbent. Specifically, insights into the distinct boron adsorption sites were obtained by solid-state one-dimensional (1D) 11B magic-angle spinning NMR and the local structures and intermolecular interactions in the vicinities of boron chelate complexes are elucidated by analyzing two-dimensional (2D) 1H-13C and 11B-1H heteronuclear correlation NMR spectra. The total boron adsorption capacity of cellulose is estimated to be in the 1.3-3.0 mg range per gram of sorbent, which is lower than the boron adsorption capacity of a polystyrene-based resin, ∼17.2 mg of boron per gram of Amberlite IRA 743. Our study demonstrates that the local backbone and side chain flexibility as well as the structures of polyol groups play a significant role in determining the kinetic and thermodynamic stability of chelate complexes, yielding to different boron adsorption capabilities of lignocellulosic polymers.

Enhanced catalytic cleavage of C-O and C-C bonds of raw biomass into lignin monomers and glucose

Reductive catalytic fractionation (RCF) of lignocellulose has been considered to be a promising strategy for the separation and subsequent upgradation of biomass constituents. However, the recalcitrant C-C bonds in lignin were the major limitation to produce aromatic monomers in high yield for the hydrogenolysis of lignocellulosic biomass under mild conditions. Herein, an efficient approach was developed to catalytic in-situ hydrogenolysis of solid biomass into lignin monomers and glucose using Ni-Al catalyst with Ni2Al3 alloy phase. 62.9 wt% of aromatic monomers yield was achieved with the degree of delignification up to 89.8 %. Two-dimensional heteronuclear single quantum correlation nuclear magnetic resonance (2D HSQC NMR) spectrum emerged that the C-O and C-C bonds involving β-O-4, β-β and β-5 linkages were all catalytically broken. Moreover, the cellulose was recovered with the cellulose retention and purity of 80.1 % and 88.2 %, respectively, which was further enzymatically hydrolyzed to glucose with the yield of 92 %. Overall, the efficient fractionation and transformation of biomass constituents were realized simultaneously.

Unequivocal identification of two-bond heteronuclear correlations in natural products at nanomole scale by i-HMBC

HMBC is an essential NMR experiment for determining multiple bond heteronuclear correlations in small to medium-sized organic molecules, including natural products, yet its major limitation is the inability to differentiate two-bond from longer-range correlations. There have been several attempts to address this issue, but all reported approaches suffer various drawbacks, such as restricted utility and poor sensitivity. Here we present a sensitive and universal methodology to identify two-bond HMBC correlations using isotope shifts, referred to as i-HMBC (isotope shift detection HMBC). Experimental utility was demonstrated at the sub-milligram / nanomole scale with only a few hours of acquisition time required for structure elucidation of several complex proton-deficient natural products, which could not be fully elucidated by conventional 2D NMR experiments. Because i-HMBC overcomes the key limitation of HMBC without significant reduction in sensitivity or performance, i-HMBC can be used as a complement to HMBC when unambiguous identifications of two-bond correlations are needed.

Molecular interactions between apigenin and starch with different amylose/amylopectin ratios revealed by X-ray diffraction, FT-IR and solid-state NMR

Starch can readily form complexes with polyphenols. However, its two components, namely amylose and amylopectin, differ significantly in their ability to complex with phenolic compounds. Given that the mechanism of their interaction is still poorly studied, this work investigated intermolecular interactions between apigenin and starch with different amylose/amylopectin ratios using 1H NMR, FT-IR, XRD, DSC and solid-state NMR. Results showed that corn starch with high amylose/amylopectin ratios had a better complexing ability and higher complexing index with apigenin than amylopectin. Besides, solid-state NMR suggested that the molecular mechanism behind the strong intermolecular interactions between corn starch and apigenin involved hydrogen bonds. Furthermore, the detailed binding sites of hydrogen bonds, that linked by hydroxyl-starch and phenyl-apigenin were also confirmed by 1H13C heteronuclear correlation (HETCOR) spectra. This study revealed the molecular mechanism on amylose/amylopectin complexing with apigenin and provides a theoretical basis for further developing polyphenols in starchy food.

Zwitterionic or Not? Fast and Reliable Structure Determination by Combining Crystal Structure Prediction and Solid-State NMR.

When it comes to crystal structure determination, computational approaches such as Crystal Structure Prediction (CSP) have gained more and more attention since they offer some insight on how atoms and molecules are packed in the solid state, starting from only very basic information without diffraction data. Furthermore, it is well known that the coupling of CSP with solid-state NMR (SSNMR) greatly enhances the performance and the accuracy of the predictive method, leading to the so-called CSP-NMR crystallography (CSP-NMRX). In this paper, we present the successful application of CSP-NMRX to determine the crystal structure of three structural isomers of pyridine dicarboxylic acid, namely quinolinic, dipicolinic and dinicotinic acids, which can be in a zwitterionic form, or not, in the solid state. In a first step, mono- and bidimensional SSNMR spectra, i.e., 1H Magic-Angle Spinning (MAS), 13C and 15N Cross Polarisation Magic-Angle Spinning (CPMAS), 1H Double Quantum (DQ) MAS, 1H-13C HETeronuclear CORrelation (HETCOR), were used to determine the correct molecular structure (i.e., zwitterionic or not) and the local molecular arrangement; at the end, the RMSEs between experimental and computed 1H and 13C chemical shifts allowed the selection of the correct predicted structure for each system. Interestingly, while quinolinic and dipicolinic acids are zwitterionic and non-zwitterionic, respectively, in the solid state, dinicotinic acid exhibits in its crystal structure a "zwitterionic-non-zwitterionic continuum state" in which the proton is shared between the carboxylic moiety and the pyridinic nitrogen. Very refined SSNMR experiments were carried out, i.e., 14N-1H Phase-Modulated (PM) pulse and Rotational-Echo Saturation-Pulse Double-Resonance (RESPDOR), to provide an accurate N-H distance value confirming the hybrid nature of the molecule. The CSP-NMRX method showed a remarkable match between the selected structures and the experimental ones. The correct molecular input provided by SSNMR reduced the number of CSP calculations to be performed, leading to different predicted structures, while RMSEs provided an independent parameter with respect to the computed energy for the selection of the best candidate.

Insight into the mechanism underlying modification of Neosinocalamus affinis by hygrothermal treatment

Abstract Hygrothermal treatment is an effective method for improving the dimensional stability of bamboo. In this study, changes in the physicochemical properties of Neosinocalamus affinis after hygrothermal treatment were comprehensively investigated: the hemicellulose content decreased, C=O in the acetyl group and the hydroxyl content decreased and xylan was partially degraded. The dimensional stability of N. affinis gradually increased with temperature, and optimal values were obtained at 220 °C, as indicated by a 16.5% decrease in anti-swelling efficiency and a 93.7% increase in contact angle. Alterations in the macromolecular structure of lignin were also observed: the contents of β-O-4 linkages and p-coumarate decreased by 54.3% and 23.9%, respectively; β-5 linkages disappeared at 220 °C, as determined by heteronuclear single quantum correlation spectroscopy. However, the maximum values for crystallinity, nano indentation elastic modulus, and hardness were reached at 180 °C and were higher than those of the untreated samples by 8.6%, 19.9%, and 23.5%, respectively. With the combined application of physical mechanics and dimensional stability, hygrothermal treatment at 180 °C and 100% relative humidity was proved to exert the optimal effects on N. affinis. These results provide new and comprehensive insights into the mechanism allowing the modification of N. affinis by hygrothermal treatment.

NMR Study on Laccase Polymerization of Kraft Lignin Using Different Enzymes Source.

The usage of laccases is a sustainable and environmentally friendly approach to modifying the Kraft lignin structure for use in certain applications. However, the inherent structure of Kraft lignin, as well as that resulting from laccase modification, still presents challenges for fundamental comprehension and successful lignin valorization. In this study, bacterial and fungal laccases were employed to modify eucalypt Kraft lignin. To evaluate the type and range of the chemical and structural changes of laccase-treated lignins, different NMR techniques, including solution 1H and 2D NMR (heteronuclear single quantum correlation (HSQC)), and solid-state 13C NMR, were applied. Size exclusion chromatography and infrared spectroscopy were also used. Interestingly, HSQC analysis showed substantial changes in the oxygenated aliphatic region of lignins, showing an almost complete absence of signals corresponding to side-chains due to laccase depolymerization. Simultaneously, a significant loss of aromatic signals was observed by HSQC and 1H NMR, which was attributed to a deprotonation of the lignin benzenic rings due to polymerization/condensation by laccase reactions. Then, condensed structures, such as α-5', 5-5', and 4-O-5', were detected by HSQC and 13C NMR, supporting the increment in molecular weight, as well as the phenolic content reduction determined in lignins.

Satellite-transition double cross-polarization HETCOR under fast MAS

Double-cross polarization to the satellite-transitions (STs) of half-integer quadrupolar nuclei is demonstrated using proton-detected heteronuclear correlation (HETCOR) under fast magic-angle spinning (MAS). By placing the rf frequency away from the central-transition (CT) and selective to the STs, average Hamiltonian theory shows a scaled effective rf field with a phase equal to the complex ST spinning sideband being irradiated. Such an effective rf field can excite and spinlock STs but the phase spread usually leads to signal cancellation in one-step excitation or cross polarization experiments. The cancellation does not occur for two-step double cross-polarization (DCP) HETCOR experiments, therefore high efficiencies can be obtained. With careful magic-angle calibration, ST and double-quantum ST (DQST) HETCOR experiments are demonstrated with the 35Cl nuclei in histidine·HCl·H2O. These experiments provide additional information over the commonly observed CT spectra and near isotropic resolution in the case of DQST of spin S = 3/2 quadrupolar nuclei.

Primary Structure of Glycans by NMR Spectroscopy.

Glycans, carbohydrate molecules in the realm of biology, are present as biomedically important glycoconjugates and a characteristic aspect is that their structures in many instances are branched. In determining the primary structure of a glycan, the sugar components including the absolute configuration and ring form, anomeric configuration, linkage(s), sequence, and substituents should be elucidated. Solution state NMR spectroscopy offers a unique opportunity to resolve all these aspects at atomic resolution. During the last two decades, advancement of both NMR experiments and spectrometer hardware have made it possible to unravel carbohydrate structure more efficiently. These developments applicable to glycans include, inter alia, NMR experiments that reduce spectral overlap, use selective excitations, record tilted projections of multidimensional spectra, acquire spectra by multiple receivers, utilize polarization by fast-pulsing techniques, concatenate pulse-sequence modules to acquire several spectra in a single measurement, acquire pure shift correlated spectra devoid of scalar couplings, employ stable isotope labeling to efficiently obtain homo- and/or heteronuclear correlations, as well as those that rely on dipolar cross-correlated interactions for sequential information. Refined computer programs for NMR spin simulation and chemical shift prediction aid the structural elucidation of glycans, which are notorious for their limited spectral dispersion. Hardware developments include cryogenically cold probes and dynamic nuclear polarization techniques, both resulting in enhanced sensitivity as well as ultrahigh field NMR spectrometers with a 1H NMR resonance frequency higher than 1 GHz, thus improving resolution of resonances. Taken together, the developments have made and will in the future make it possible to elucidate carbohydrate structure in great detail, thereby forming the basis for understanding of how glycans interact with other molecules.

Multiblock poly(ether-b-amide) copolymers comprised of PA1212 and PPO-PEO-PPO with specific moisture-responsive and antistatic properties

Functional multiblock poly(ether-b-amide) (PEBA) copolymers, comprised of PA1212 (polyamide 1212) as hard segments and Jeffamine ED-2003 as soft segments, were successfully prepared via two-step melt polycondensation without any amidation catalyst. Here, using diamino-terminated poly(propylene oxide)-poly(ethylene oxide)-poly(propylene oxide) (PPO-PEO-PPO), Jeffamine ED-2003, enhances the compatibility between polyamide oligomer and polyether, which is better than the traditional route using hydroxyl-terminated polyether. The chemical structure of multiblock PEBAs, as well as the microphase separated structure with crystalline phase of polyamide and polyether, were confirmed by heteronuclear multiple-bond correlation spectrum, heteronuclear multiple quantum correlation spectrum, Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry and dynamic mechanical analysis. The hydrophilic PEBA copolymers showed water adsorption ranging from 87.3% to 17.1% depending on the polyether content, and specially showed moisture responsive behavior within seconds when exposed to moisture. The corresponding mechanism was studied using time-resolved attenuated total reflectance FT-IR spectroscopy in the molecular level and the water diffusion coefficient was estimated to be 1.07 × 10–8 cm2∙s–1. Two-dimensional correlation FT-IR spectra analysis was performed to confirm that the interaction between water and polyether phase was in preference to that between water and polyamide matrix, and water molecule only forms hydrogen bond with the polyether segment. Due to the incorporation of PEO segments, the PEBAs have the surface resistivity varying from 5.6 × 109 to 6.5× 1010 Ω, which makes PEBA potential candidate as permanent antistatic agent.

An investigation into the adsorption mechanism of n-butanol by ZIF-8: a combined experimental and ab initio molecular dynamics approach.

The zeolitic imidazolate framework, ZIF-8, has been shown by experimental methods to have a maximum saturation adsorption capacity of 0.36 g g-1 for n-butanol from aqueous solution, equivalent to a loading of 14 butanol molecules per unit cell or 7 molecules per sodalite β-cage. Diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) shows the presence of hydrogen bonding between adsorbed butanol molecules within the cage; the presence of three different O-H stretching modes indicates the formation of butanol clusters of varying size. Ab initio molecular dynamics simulations show the formation of intermolecular hydrogen bonding between the butanol molecules, with an average hydrogen-bond coordination number of 0.9 after 15 ps simulation time. The simulations also uniquely demonstrate the presence of weaker interactions between the alcohol O-H group and the π-orbital of the imidazole ring on the internal surface of the cage during early stages of adsorption. The calculated adsorption energy per butanol molecule is -33.7 kJ mol-1, confirming that the butanol is only weakly bound, driven primarily by the hydrogen bonding. Solid-state MAS NMR spectra suggest that the adsorbed butanol molecules possess a reasonable degree of mobility in their adsorbed state, rather than being rigidly held in specific sites. 2D 13C-1H heteronuclear correlation (HETCOR) experiments show interactions between the butanol aliphatic chain and the ZIF-8 framework experimentally, suggesting that O-H interactions with the π-orbital are only short lived. The insight gained from these results will allow the design of more efficient ways of recovering and isolating n-butanol, an important biofuel, from low-concentration solutions.

A Reference-Free Calibration for Ultrafast Spatiotemporally Encoded 2-D NMR Spectroscopy

Spatiotemporal encoding (SPEN) is a state-of-the-art nuclear magnetic resonance (NMR) technique, utilized to acquire 2-D or even multidimensional ( <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$n$ </tex-math></inline-formula> -D) NMR spectra and/or images in a single (or a few) scans. Benefitting from special encoding and decoding schemes, SPEN can thus accelerate the classical 2-D NMR experiment by orders of magnitude. SPEN’s decoding in particular is executed under the effects of oscillating magnetic field gradients; inevitable hardware imperfections may then give rise to inconsistencies between odd and even SPEN data lines, making only half (odd or even) of the acquired data available for processing. This halves the spectral bandwidth available in the direct dimension and decreases the spectral sensitivity, leading to the emergence of peak folding artifacts and/or a decline of the spectral quality. Past research has been carried out to correct these distortions induced by gradient imperfections, but requires an extra reference scan collected under identical experimental conditions. This work presents a detailed theoretical analysis of the impacts of various imperfections on the 2-D SPEN NMR spectrum and proposes a three-step reference-free calibration algorithm in accordance with theoretical analysis. Experimental data on single-shot 2-D correlation spectroscopy (COSY) and heteronuclear single-quantum correlation (HSQC) NMR spectra collected using SPEN were performed and processed, which demonstrate the feasibility and applicability of the proposed reference-free algorithm.

One-pot synthesis of liquid photocrosslinkable poly(l-lactide) with terminal triacrylate.

We synthesized a poly(l-lactide)-pentaerythritol triacrylate (PETA) polymer modified with acrylic trifunctional groups using a one-pot method based on ring-opening polymerization of l-lactide and PETA. We calculated the molecular weight and structure of PLA-PETA using gel permeation chromatography (GPC) and nuclear magnetic resonance (NMR) (1H, 13C, heteronuclear multiple bond correlation [HMBC]) spectroscopy. Photocrosslinking PLA-PETA using the Omnirad 1173 photoinitiator yielded a transparent sample with 91% crosslinkage. The crosslinked sample was analyzed using differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and thermomechanical analysis (TMA) to determine its thermal properties and thermal expansion coefficient. In vitro cell toxicity tests showed an average cell viability >90%, indicating that the PLA-PETA polymer had good biocompatibility with cells after photocrosslinking.

NMR Titration Studies in Z-DNA Dynamics.

Chemical shift perturbation (CSP) is a simple NMR technique for studying the DNA binding of proteins. Titration of the unlabeled DNA into the 15N-labeled protein is monitored by acquiring a two-dimensional (2D) heteronuclear single-quantum correlation (HSQC) spectrum at each step of the titration. CSP can also provide information on the DNA-binding dynamics of proteins, as well as protein-induced conformational changes in DNA. Here, we describe the titration of DNA for the 15N-labeled Z-DNA-binding protein, monitored via 2D HSQC spectra. NMR titration data can be analyzed with the active B-Z transition model to provide the protein-induced B-Z transition dynamics of DNA.

Enhanced production of sugars and UV-shielded lignin/PAN fiber mats from chemi-mechanical pulps

This study investigated poplar pretreatments by chemi-mechanical pulping (CMP) under different beating degrees and alkali concentrations. The enzyme-mediated hydrolysis of pretreated poplar was enhanced by deacetylation and delignification. Meanwhile, the remaining lignin residues were used to produce lignin/polyacrylonitrile (PAN) fiber mats by electrospinning. These mats exhibited excellent mechanical and UV-blocking performance when the lignin was obtained from pulps under milder alkali concentrations (5 g/L). 31P nuclear magnetic resonance (31P NMR) and two-dimensional heteronuclear single-quantum correlation nuclear magnetic resonance (2D HSQC NMR) data revealed that increasing the beating degree at low alkali concentration during the CMP process led to the cleavage of β-O-4′ interunit linkages and re-condensation in lignin, releasing several phenolic groups. Lignin with more linear β-O-4′ interunit linkages and lesser phenolic groups, obtained from treatment of CMP with lower alkali concentration (5 g/L) and beating degree (20°SR), resulted in the corresponding lignin/PAN fiber mats exhibiting better mechanical performance. Further, lignin, along with the increased phenolic-OH and COOH, and p-hydroxybenzoate (PB) units with a more extended conjugate structure, derived from CMP under lower alkali concentration (5 g/L) and higher beating degree (45°SR), led to a stronger ultraviolet (UV) absorption in the corresponding lignin/PAN mats. To summarize, this study reports a mild and low-pollution biomass pretreatment method (CMP) that can efficiently regulate the lignin structure and exhibit efficient anti-ultraviolet properties. The corresponding UV-blocking fiber mats can be potentially used as materials for wearable fabrics.