Robust Oxidase-Mimetic Supramolecular Nanocatalyst for Lignin Biodegradation.

Enzymatic catalysis presents an eco-friendly, energy-efficient method for lignin degradation. However, challenges arise due to the inherent incompatibility between enzymes and native lignin. In this work, we introduce a supramolecular catalyst composed of fluorenyl-modified amino acids and Cu2+, designed based on the aromatic stacking of the fluorenyl group, which can operate in ionic liquid environments suitable for the dissolution of native lignin. Amino acids and halide anions of ionic liquids shape the copper site's coordination sphere, showcasing remarkable catechol oxidase-mimetic activity. The catalyst exhibits thermophilic property, and maintains oxidative activity up to 75 °C, which allows the catalyzed degradation of the as-dissolved native lignin with high efficiency even without assistance of the electron mediator. In contrast, at this condition, the native copper-dependent oxidase completely lost its activity. This catalyst with superior stability and activity offer promise for sustainable lignin valorization through biocatalytic routes compatible with ionic liquid pretreatment, addressing limitations in native enzymes for industrially relevant conditions.

Histidine-Based Supramolecular Nanoassembly Exhibiting Dual Enzyme-Mimetic Functions: Altering the Tautomeric Preference of Histidine to Tailor Oxidative/Hydrolytic Catalysis.

Challenges persist in replicating enzyme-like active sites with functional group arrangements in supramolecular catalysis. In this study, we present a supramolecular material comprising Fmoc-modified histidine and copper. We also investigated the impact of noncanonical amino acids (δmH and εmH), isomers of histidine, on the catalytic process. The Fmoc-δmH-based nanoassembly exhibits an approximately 15-fold increase in oxidative activity and an ∼50-fold increase in hydrolytic activity compared to Fmoc-εmH (kcat/Km). This distinction arises from differences in basicity and ligation properties between the ε- and δ-nitrogen of histidine. The addition of guanosine monophosphate further enhances the oxidative activity of the histidine- and methylated histidine-based catalysts. The Fmoc-δmH/Cu2+-based nanoassembly catalyzes the oxidation/hydrolysis cascade of 2',7'-dichlorofluorescein diacetate, benefiting from the synergistic effect between the copper center and the nonligating ε-nitrogen of histidine. These findings advance the biomimetic catalyst design and provide insights into the mechanistic role of essential residues in natural systems.

Residual Change of Four Pesticides in the Processing of Pogostemon cablin and Associated Factors.

Before use as medicines, most traditional Chinese medicine (TCM) plants are processed and decocted. During processing, there may be some changes in pesticide residues in TCM. In recent years, reports have studied the changes of pesticides during the processes of boiling, drying and peeling of TCM materials but have rarely involved special processing methods for TCM, such as ethanol extraction and volatile oil extraction. The changes of carbendazim, carbofuran, pyridaben and tebuconazole residues in common processing methods for P. cablin products were systemically assessed in this study. After each processing step, the pesticides were quantitated by UPLC-MS/MS. The results showed amount decreases in various pesticides to different extents after each processing procedure. Processing factor (PF) values for the four pesticides after decoction, 75% ethanol extraction and volatile oil extraction were 0.02~0.75, 0.40~0.98 and 0~0.02, respectively, which indicated that residual pesticide concentrations may depend on the processing technique. A risk assessment according to the hazard quotient with PF values showed that residual pesticide amounts in P. cablin were substantially lower than levels potentially posing a health risk. Overall, these findings provide insights into the safety assessment of P. cablin.

Enzyme-Mimicking Materials from Designed Self-Assembly of Lysine-Rich Peptides and G-Quadruplex DNA/Hemin DNAzyme: Charge Effect of the Key Residues on the Catalytic Functions.

In enzymatic active sites, the essential functional groups are spatially arranged as a result of the enzyme three-dimensional folding, which leads to remarkable catalytic properties. We are inspired to self-assemble the polylysine peptides with guanine-rich DNA and hemin as cofactor to fabricate the peroxidase-mimicking catalytic nanomaterials. The DNA can fold into G-quadruplex to provide a supramolecular scaffold and a nucleobase for supporting and coordinating hemin, and the polylysine provides amine as distal groups to promote the H2O2 adsorption to the iron of hemin. The polylysine and DNA components synergistically accelerated the hemin-catalyzed reactions, and the complex containing ε-polylysine exhibited higher activity than α-polylysine. This activity difference is attributed to the higher pKa value and more susceptible protonation of amine of ε-polylysine than α-polylysine. The ε-polylysine/DNA/hemin had similar coordination states of hemin and conformations of the components to α-polylysine/DNA/hemin but accelerated the formation of the intermediate compound I faster than α-polylysine. Theoretical simulation reveals that the unprotonated NH2 behaved like a base catalyst, similar to His-42 residue in the natural heme pocket, while the protonated NH3+ acted as an acid, which indicated that the base catalyst on the distal side of the hemin pocket is more active than the acid. This work provides an avenue to control the distribution of the catalytic residues in an enzyme-like active site and to understand the roles of the key residues of native enzymes.

Heme-Dependent Supramolecular Nanocatalysts: A Review.

Enzymes fold into three-dimensional structures to distribute amino acid residues for catalysis, which inspired the supramolecular approach to construct enzyme-mimicking catalysts. A key concern in the development of supramolecular strategies is the ability to confine and orient functional groups to form enzyme-like active sites in artificial materials. This review introduces the design principles and construction of supramolecular nanomaterials exhibiting catalytic functions of heme-dependent enzymes, a large class of metalloproteins, which rely on a heme cofactor and spatially configured residues to catalyze diverse reactions via a complex multistep mechanism. We focus on the structure-activity relationship of the supramolecular catalysts and their applications in materials synthesis/degradation, biosensing, and therapeutics. The heme-free catalysts that catalyze reactions achieved by hemeproteins are also briefly discussed. Towards the end of the review, we discuss the outlook on the challenges related to catalyst design and future prospective, including the development of structure-resolving techniques and design concepts, with the aim of creating enzyme-mimicking materials that possess catalytic power rivaling that of natural enzymes..

Colorimetric Sensor Based on the Oxidase-Mimic Supramolecular Catalyst for Selective and Sensitive Biomolecular Detection.

We have engineered a colorimetric sensor capable of selective and sensitive detection of amino acids. This sensor employs a supramolecular copper-dependent oxidase mimic as the probe, stemming from our prior research. The oxidase mimic is constructed through the self-assembly of commercially available guanosine monophosphate (GMP), Fmoc-lysine, and Cu2+. It catalyzes the formation of a red product with a maximum absorbance at 510 nm. The changes in color and absorbance are responsive to both the concentrations and types of amino acids present. This effect is most pronounced in the presence of histidine, with a detection limit (LOD) of 6.4 nM. Furthermore, the catalytic probe can distinguish histidine from histamine and imidazole propionate, as well as 1-methyl-histidine from 3-methyl-histidine, based on their distinct coordination capacities with copper. This underscores the high selectivity of the sensing platform. Both theoretical simulations and experimental results (including UV-vis spectra, fluorescence, and EPR) indicate that the amino acids may engage in copper center coordination, thereby impeding O2 access to copper─a pivotal aspect of the oxidase catalysis. This sensing platform, characteristic of its swift response, simple fabrication, and exceptional sensitivity and selectivity, can also be applied to detect other biological analytes such as nucleotides. It holds potential for use in environmental and biochemical analyses.

A designed DNA/amino acid amphiphile-based supramolecular oxidase-mimetic catalyst for colorimetric DNA detection.

DNA is self-assembled with Fmoc-amino acids and Cu2+ to construct a supramolecular catechol oxidase-mimetic catalyst, which exhibits remarkable activity in catalyzing colorimetric reactions. This catalytic system is used for the detection of DNA hybridization with a high selectivity and a low detection limit.

Seasonal variation in δ13C of Pinus. yunnanensis and Pinus. armandii at different stand ages.

Seasonal drought is common in Yunnan province, and water is the dominant factor limiting the growth of Pinus. yunnanensis and Pinus. armandii. The water use efficiency (WUE) of the two species is poorly understood. Needles were collected in a plantation (P. yunnanensis and P. armandii mixed forest) in four seasons, and the needle δ13C values were measured. The selected species had larger δ13C values and exhibited higher WUE than typical subtropical species. P. armandii needles showed a more conservative water use strategy (high WUE) than P. yunnanensis. There were significant differences in the δ13C values of P. armandii between the two ages, whereas no difference was observed in the δ13C values of P. yunnanensis. The lowest δ13C value in the young P. armandii forest was observed in spring, whereas the δ13C value of middle-aged forests did not differ between the seasons. The δ13C value of young P. yunnanensis forests showed no difference in the four seasons, and the maximum value was observed in summer in middle-aged forests. In general, the δ13C value of P. armandii was lowest in spring, whereas that of P. yunnanensis was higher in spring and winter. The needle δ13C values were lower in spring and winter, indicating that the season had different effects on the δ13C values of different tree species. Correlation analysis between the needle δ13C values and meteorological data indicated that temperature and precipitation were the dominant factors affecting WUE in P. yunnanensis and P. armandii. The effect of temperature on WUE was greater in P. yunnanensis middle-aged forests. The identification and selection of subtropical tree species with high WUE are critical to maintaining high levels of forest benefits under limited water conditions.

Seasonal variation in C:N:P stoichiometry, nonstructural carbohydrates, and carbon isotopes of two coniferous pioneer tree species in subtropical China.

The characteristics of C:N:P stoichiometry, nonstructural carbohydrate (NSC) content, and C stable isotopes and their relationships affect plant responses to environmental changes and are critical to understanding the ecosystem carbon and water cycles. We investigated the water use strategies and physiological changes of two pioneer tree species (Pinus armandii and Pinus yunnanensis) in response to seasonal drought in subtropical China. The seasonal variation in needle δ13C values, C:N:P stoichiometry, and NSC contents of the two tree species were studied in 25-year-old plantation in central Yunnan Province. The needle δ13C values of both species were highest in summer. Soluble sugars, starch and NSC content of the two tree species decreased from spring to winter, while there was no significant difference in the seasonal variation of soluble sugars/starch in P. armandii needles, the maximum soluble sugars/starch in P. yunnanensis needles was in autumn. In addition, the C, N, and P contents of the needles and the C:N and C:P ratios of the two species showed different seasonal fluctuations, whereas the N:P ratio decreased with the season. The C:N:P stoichiometry and NSC content of the needles showed significant correlations, whereas the needle δ13C was weakly correlated with C:N:P stoichiometry and NSC content. Phenotypic plasticity analysis and principal component analysis revealed that the needle nutrient characteristics (NSC and P contents and N:P ratio) and needle δ13C values were critical indicators of physiological adaptation strategies of P. armandii and P. yunnanensis for coping with seasonal variation. These results increase our understanding of the water-use characteristics of the two pioneer tree species and the dynamic balance between the NSC, C, N, and P contents of the needles.

A nucleotide-copper(II) complex possessing a monooxygenase-like catalytic function.

The de novo design of artificial biocatalysts with enzyme-like active sites and catalytic functions has long been an attractive yet challenging goal. In this study, we present a nucleotide-Cu2+ complex, synthesized through a one-pot approach, capable of catalyzing ortho-hydroxylation reactions resembling those of minimalist monooxygenases. Both experimental and theoretical findings demonstrate that the catalyst, in which Cu2+ coordinates with both the nucleobase and phosphate moieties, forms a ternary-complex intermediate with H2O2 and tyramine substrates through multiple weak interactions. The subsequent electron transfer and hydrogen (or proton) transfer steps lead to the ortho-hydroxylation of tyramine, where the single copper center exhibits a similar function to natural dicopper sites. Moreover, Cu2+ bound to nucleotides or oligonucleotides exhibits thermophilic catalytic properties within the temperature range of 25 °C to 75 °C, while native enzymes are fully deactivated above 35 °C. This study may provide insights for the future design of oxidase-mimetic catalysts and serve as a guide for the design of primitive metallocentre-dependent enzymes.

Low-Temperature Fibre Direct Compounding of Cellulose Fibres into PA6.

This study reports on the development of a novel polymer processing approach that combines low-temperature (LT) processing and fibre direct compounding (FDC) to reduce the thermal stress on thermosensitive components that occurs during compounding and subsequent injection moulding (IM). Composites based on polyamide 6 (PA6) and cellulose fibres (CeF) were prepared using an LT-FDC process and in parallel with a conventional approach using a twin-screw extruder and IM. The morphological, optical, thermal, and mechanical properties of the prepared samples were investigated using optical microscopy (OM), differential scanning calorimetry (DSC), colorimetry, dynamic mechanical analysis (DMA) and tensile tests. Composites prepared using LT-FDC exhibited worse fibre dispersion but lower fibre degradation. In comparison to neat PA6, the LT-FDC composites had increased tensile modulus (Et) and storage modulus (E') at 120 °C by up to 32% and 50%, respectively, while the tensile strength (σm) decreased by 20%.

Switchable Enzyme-mimicking catalysts Self-Assembled from de novo designed peptides and DNA G-quadruplex/hemin complex.

Reconstruction of enzymatic active site in an artificial system is key to achieving high catalytic efficiency. Herein, we report the self-assembly of the lysine-containing peptides with guanine-rich DNA and hemin to form peroxidase-mimicking active sites and catalytic nanoparticles. The DNA strand self-folds into a G-quadruplex structure that provides a supramolecular scaffold and a potential axial ligand for hemin. The ß-sheet forming capability of the lysine-containing peptides is found to affect the catalytic synergy between the G-quadruplex DNA and the peptide. It is hypothesized that the ß-sheet formation of the peptides results in the enrichment of the lysine residues, which distribute on the distal side of hemin to promote the formation of Compound I, like distal arginine residue in natural heme pocket. Incorporation of the histidine residues into the lysine-containing peptides further enhanced the hemin activities, indicating the cooperation between the lysine and histidine. Furthermore, the peptide/DNA/hemin complexes can be switched between active and inactive state by reversible formation and deformation of the DNA G-quadruplex, which was attributed to the peptides-promoted conformational changes of the DNA components. This work opens an avenue to mimic the catalytic residues and their spatial distribution in the natural enzymes, and shed light on the design of the smart biocatalysts that can respond to the environmental stimuli.

Automated measurement of endometrial peristalsis in cine transvaginal ultrasound images.

Objectives: Endometrial peristalsis (EP) in non-pregnant uterine can be assessed by visual assessment of transvaginal ultrasound (TVUS). However, visual assessment is subjective, and the outcome depends on the sonographers and video analysts. This study aimed to create a newly developed automatic analysis algorithm for measuring the EP compared to visual assessment. Methods: A retrospective analysis was performed using the datasets from in vitro fertilization and embryo transfer (IVF-ET), who underwent the evaluation of EP by TVUS within 5 days prior to transplantation. 158 cine TVUS images were used to develop the automated analysis algorithm, and 37 cine TVUS images were evaluated by both visual and automated analysis algorithms. The algorithm was developed by applying the optical flow technology and enabled objective analysis of the number, direction, and intensity of EP. Results: The number of peristaltic waves counted by visual assessment was 4.2 ± 2.3 (mean ± standard deviation) and 4.1 ± 2.1 for doctors one and two, respectively. The number of waves counted with the algorithm was 3.6 ± 2.1 at first evaluation and 3.7 ± 2.0 at repeated evaluation. A significant difference was found between the algorithm count and visual assessment (p = 0.001, 0.002, 0.003, 0.008). The ICC values for algorithm versus manuals ranged from 0.84 to 0.96 and 0.87 to 0.96. The numbers of the cervix-to-fundus (CF), fundus-to-cervix (FC), and both cervix-to-fundal and fundus-to-cervix (CF + FC) directions of EP counted by the algorithm were 50, 52, and 32, respectively. The numbers counted by visual assessment were 43, 45, and 46, respectively. The number of EP was the same in 87% of the two algorithm counts. The number was lower between the algorithm and visual analysis (79% with complete agreement). The EP intensity assessed by the algorithm was 2.6 ± 1.1, and the peristalsis velocity was 0.147 (0.07) mm/s. Conclusion: The fully automated analysis algorithm can be used to quantify uterine peristalsis comparable to visual assessment.

Levels and Health Risk of Pesticide Residues in Chinese Herbal Medicines.

In the present study, 168 pesticides in 1,017 samples of 10 Chinese herbal medicines (CHMs) were simultaneously determined by high-performance liquid (HPLC-MS/MS) and gas (GC-MS/MS) chromatography-tandem mass spectrometry. A total of 89.2% of the samples encompassed one or multiple pesticide residues, and the residue concentrations in 60.5% of samples were less than 0.02 mg kg-1, revealing the relatively low residue levels. The hazard quotient and hazard index methods were used to estimate the health risk for consumers. For a more accurate risk assessment, the exposure frequency and exposure duration of CHMs were involved into the exposure assessment, which was obtained from a questionnaire data of 20,917 volunteers. The results of chronic, acute, and cumulative risk assessment indicated that consumption of CHMs is unlikely to pose a health risk to consumers. Ranking the risk of detected pesticides revealed that phorate, BHC, triazophos, methidathion, terbufos, and omethoate posed the highest risk. Our results also showed that pollution of the aboveground medicinal part was more serious. Although exposure to pesticides in tested CHMs was below dangerous levels, more strict controlled management should be carried out for banned pesticides due to the high detection rate and illegal use in the actual planting practice.