Transformants Research Articles

Organophotocatalyst Enabled Deoxycyclopropanation of Alcohols.

Cyclopropane fragments, which widely exist in marketed drugs and natural products, can confer special pharmacological properties to small-molecule drugs. Therefore, developing methods to construct cyclopropanes is of great significance. Nevertheless, the introduction of cyclopropane primarily relies on already-formed cyclopropyl groups, which significantly restricts the diversity of cyclopropane skeletons. Late-stage direct cyclopropanation is still a challenging task. Herein, a photo-induced intermolecular deoxycyclopropanation reaction that employs alcohols as substrates, and 1 mol.% of 2,3,5,6-tetrakis(carbazol-9-yl)-1,4-dicyanobenzene (4CzTPN)as organophotocatalyst is reported. This method proceeds with high transformation efficiency (up to 98% yield) and exhibits broad functional group tolerance, such as primary, secondary, and tertiary alcohols as well as various activated β-halogenated alkenes. This process is mild, easy to operate, and has low equipment requirements. The power of this technology is demonstrated by the late-stage functionalization of five marketed drugs and five natural products.

Enhancing Agrobacterium-mediated soybean transformation efficiency with an auxiliary solution

Soybean is a crucial source of oil, protein, and biofuel, necessitating efficient transformation systems for advancing research. Agrobacterium-mediated transformation is currently the primary method used in the soybean transformation industry and scientific research. However, the low efficiency and genotype dependency of this technology leave significant room for improvement. This study aimed to enhance soybean transformation efficiency by generating and validating three reporter vectors (ZsGreen, TdTomato, and Ruby) and using Agrobacterium Auxiliary Solution (AAS) containing Silwet L-77 and hormone mixtures. Our findings demonstrate that AAS significantly improves hairy root transformation rates. Specifically, this combination increased total root and cotyledon transformation efficiencies compared to the control. We also found that larger vectors like Ruby reduced transformation efficiency compared to smaller markers like GFP and RFP. Furthermore, AAS slightly reduced the co-transformation rate of two separate vectors compared to single vector transformations. Additionally, AAS enhanced soybean hypocotyl transformation rates across various varieties, consistently increasing positive root and explant efficiencies. Notably, transformation rates varied significantly between varieties, with Forrest differing from Williams 82 and Dongnong 50. This research highlights the importance of auxiliary agents and vector size in optimizing soybean transformation, providing insights for future advancements in genetic modification and biotechnology.

Biochemical characterization of a recombinant laccase from Halalkalibacterium halodurans C-125 and its application in the biotransformation of organic compounds.

This study aimed to produce an engineered recombinant laccase from extremophilic Halalkalibacterium halodurans C-125 (Lac-HhC-125) with higher protein yield, into a more active conformation and with properties that meet the fundamental needs of biotechnological application. The rLac-HhC125 was partially purified by size exclusion chromatography and concentrated by ultrafiltration (10kDa) with a yield of 57.6%. Oxidation reactions showed that adding 2mM CuSO4 to the assay solution led to activating the laccase. To increase its initial activity, the rLac-HhC125 was treated at 50°C for 20min before the assays, improving its performance by fourfold using the syringaldazine as a substrate. When treated with EDTA, methanol, ethanol, and DMSO, the rLac-HhC125 maintained more than 80% of its original activity. Interestingly, the acetonitrile induced a twofold activity of the rLac-HhC125. The putative rLac-HhC125 demonstrated a capability of efficient transformation of different organic compounds at pH 6, known as dye precursors, into coloured molecules. The rLac-HhC125 was active at high temperatures and alkaline pH, exhibited tolerance to organic solvents, and efficiently transformed different hydroxy derivatives into coloured compounds, which indicates that it can be used in various biotechnological processes.

The Use of the Data Envelopment Analysis–Malmquist Approach to Measure the Performance of Digital Transformation in EU Countries

Currently, the process of the digital transformation of EU countries is very important and often discussed. It will not only bring new opportunities for companies and the broader population but will also enable the transition to a more ecological economy. An important goal is to speed up the digitalization processes taking place in companies. It is very important to use already established digitalization elements more efficiently. This also resulted in the motivation for the given research. The aim of this paper is to quantify the change in the efficiency of the digital transformation of EU countries. As part of this research, the Variable Returns to Scale Data Envelopment Analysis (VRS DEA) model and the Malmquist index (MI) based on the DEA approach were applied. The results of the model made it possible to assess how the changes in technical efficiency and technological changes contributed to the changes in efficiency. The long-term theoretical added value of this paper lies in its proposal for countries and their governments to monitor not only the number of introduced digital elements, but also the efficiency of their use relative to some aggregate output; for example, GDP (Gross Domestic Product) or unemployment rate. The added value of this research is that less developed countries use digitalization elements more effectively than developed countries.

Topology of Gemini-shaped Hexagonal Heterojunction for Efficient Stereoconvergent Transformation via Dynamic Kinetic Resolution.

Despite recent advances in the combination of kinetic resolution and racemization for efficient stereoconvergent transformation, the poor stability and limited reaction activities of the products restrict their wide application in industrial production. To overcome these problems, Gemini-shaped hexagons with para-heterojunctions for one-dimensional and two-dimensional supramolecular polymers were designed via hydrogen-bonding adhesion by racemization catalyst 1and kinetic resolution 2in this work. The polymers from the assembly of Gemini-shaped hexagons exhibit rapid catalytic behaviour with efficient selectivity for the desired configuration in the synthesis of tertiary alcohols with contiguous stereocenters through dynamic kinetic resolution for the nanoscale heterojunctions of dissimilar catalysts. Among them, the developed 2D polymers gave outstanding enantioselectivities and diastereoselectivities (>99% ee, 20:1 dr) through the cooperation of adjacent dissimilar catalysts. The heterojunctions varying dimensions and distances of dissimilar catalysts provide new insight for increasing the enantioselectivity of chiral organocatalysts.

Chemoselective Pd-Based Dyotropic Rearrangement: Fluorocyclization and Regioselective Wacker Reaction of Homoallylic Amides.

Fluorocyclization of alkenes tethered with a pronucleophile is an efficient transformation that converts easily accessible starting materials to fluorinated heterocycles in a single step. We report herein an unprecedented Pd(II)-catalyzed oxidative domino process that transforms homoallylic amides to 5,6-dihydro-4H-1,3-oxazines through a domino oxypalladation/PdII-oxidation/dyotropic rearrangement/reductive elimination sequence. Three chemical bonds are created under these operationally simple conditions. Taking advantage of the facile hydrolysis of the α-fluoro tertiary alkyl ether under acidic conditions, a one-pot conversion of homoallylic amides to homologated ketones is subsequently developed, which represents a rare example of regioselective Wacker oxidation reaction of 1,1-disubstituted alkenes.

An Efficient System for Agrobacterium-Mediated Transformation of Elite Cultivars in Brassica juncea

Efficient genetic transformation approaches play pivotal roles in both gene function research and crop breeding. However, stable transformation in mustard, particularly for different horticultural types, has not been systematically studied and well-established so far. In this study, we optimized the key factors in the genetic transformation of mustard, including the optical density value of Agrobacterium suspension, the age of explants, and the combination of phytohormones at different concentrations. As a result, the optimal conditions for the genetic transformation of leaf and stem mustard included hypocotyl explants derived from 4-day-old seedlings, infection by 0.8 OD600nm Agrobacterium suspension, and then re-differentiation on a medium containing 4 mg/L trans-zeatin (tZ) and 0.1 mg/L indoleacetic acid (IAA) for leaf mustard, and for stem mustard, re-differentiation on a medium containing 2 mg/L tZ and 0.4 mg/L IAA, with positive rates of 4.74% and 5.26%, respectively. Those for root mustard were hypocotyl explants derived from 8-day-old seedlings, infection by 0.2 OD600nmAgrobacterium suspension, and a medium containing 2 mg/L tZ and 0.1 mg/L IAA, with a positive rate of 4.42%. Overall, this work provides an effective tool for both the theoretical study and genetic improvement of Brassica juncea.

Topology of Gemini‐shaped Hexagonal Heterojunction for Efficient Stereoconvergent Transformation via Dynamic Kinetic Resolution

Despite recent advances in the combination of kinetic resolution and racemization for efficient stereoconvergent transformation, the poor stability and limited reaction activities of the products restrict their wide application in industrial production. To overcome these problems, Gemini‐shaped hexagons with para‐heterojunctions for one‐dimensional and two‐dimensional supramolecular polymers were designed via hydrogen‐bonding adhesion by racemization catalyst 1 and kinetic resolution 2 in this work. The polymers from the assembly of Gemini‐shaped hexagons exhibit rapid catalytic behaviour with efficient selectivity for the desired configuration in the synthesis of tertiary alcohols with contiguous stereocenters through dynamic kinetic resolution for the nanoscale heterojunctions of dissimilar catalysts. Among them, the developed 2D polymers gave outstanding enantioselectivities and diastereoselectivities (>99% ee, 20:1 dr) through the cooperation of adjacent dissimilar catalysts. The heterojunctions varying dimensions and distances of dissimilar catalysts provide new insight for increasing the enantioselectivity of chiral organocatalysts.

Leveraging Unsupervised Task Adaptation and Semi‐Supervised Learning With Semantic‐Enriched Representations for Online Sexism Detection

ABSTRACTOver the past decade, the proliferation of hateful and sexist content targeting women on social media has become a concerning issue, adversely affecting women's lives and freedom of expression. Previous efforts to detect online sexism have utilized monolingual ensemble transformers combined with data augmentation techniques that incorporate related‐domain data, such as hate speech. However, these approaches often struggle to capture the full diversity and complexity of sexism due to limitations in the size and quality of training data. In this study, we introduce a novel sexism detection system that employs in‐domain unlabeled data through unsupervised task‐adaptation techniques and semi‐supervised learning, using an efficient single multilingual transformer model. Additionally, we incorporate a Sentence‐BERT layer to enhance our system with semantically meaningful sentence embeddings. Our proposed system outperforms existing state‐of‐the‐art methods across all tasks and datasets, demonstrating its effectiveness in detecting and addressing sexism in social media text. These results underscore the potential of our approach, providing a foundation for further research and practical applications.

Optimising Transformation Efficiency in Borrelia: Unravelling the Role of the Restriction-Modification System of Borrelia afzelii and Borrelia garinii

Borrelia spp. are transmitted to humans by the bite of an infected tick. In Europe, Borrelia afzelii and Borrelia garinii are the main causative agents of Lyme borreliosis, one of the most prevalent tick-borne diseases in the northern hemisphere. In bacteria such as Borrelia spp., a restriction-modification system (RMS) protects against the harmful introduction of foreign DNA. The RMS comprises two activities: methyltransferase and endonuclease. This study is aimed to characterize the RMS of B. afzelii and B. garinii. First, we identified potential RMS genes. The predicted genes were cloned into a methylase-deficient Escherichia coli strain and digested with methylation-sensitive restriction enzymes to verify methyltransferase activity. Additionally, the RMS proteins were purified to evaluate endonuclease activity. Subsequently, methylated and unmethylated plasmids were used to investigate the effect of methylation on endonuclease activity and transformation efficiency. We identified four possible RMS genes in B. afzelii and four RMS genes in B. garinii. We analyzed the presence of these genes in patient isolates and observed a high degree of heterogeneity. The restriction pattern of DNA methylated by each of the four recombinantly expressed genes provided strong evidence that all encode adenine-specific methyltransferases. After 24 h of incubation with purified RMS proteins, we observed complete digestion of unmethylated plasmid DNA, demonstrating endonuclease activity. Finally, we proved that methylation protects against endonuclease activity and increases transformation efficiency.

The development of an in vitro floral culture transformation system for quinoa

AbstractBecause of its high-quality seed protein and ability to thrive in marginal habitats, Chenopodium quinoa has been identified as an important emerging grain crop for global food security. However, the lack of an efficient and robust transformation system has been a barrier for conducting the advanced genetic studies needed to better understand and improve the species for agronomic traits. Here we present a novel transformation system based on Agrobacterium-mediated transformation of in vitro floral culture. Quinoa floral cultures were established from inflorescences that naturally formed on plants grown in vitro. When placed on a cytokinin-containing medium, chopped inflorescences rapidly generated highly meristematic floral cultures, primarily composed of floral buds, flowers at various developmental stages, inflorescence shoots, and leafy structures. Transformation of these cultures with Agrobacterium carrying selectable and visual markers (NPTII and GUS) produced independent, stably transformed meristematic cultures resistant to paromomycin after an extended selection period (about 3 mo with sub-culture occurring every 15 d). Transformation frequency was about 20% and was calculated as the number of independent transformed events per initial number of floral culture explants used for transformation. In vitro flowers and inflorescences from putative transgenic events self-pollinated naturally and produced viable seeds that germinated without dormancy. We also demonstrated that flowering shoots could be successfully grafted onto wild rootstock to increase the number of seeds generated from T0 floral shoots. Molecular and phenotypic analysis of the progeny confirmed that the transgenes were stably integrated and inherited according to expected Mendelian ratios.

Effect of Nucleophile Deprotonation on SuFEx: Synthesis of Sulfonamides

In a quest for efficient SuFEx-type transformations we studied reactions of amines with 4-fluorobenzenesulfonyl fluoride. The substrate may react by fluoride substitution at the aromatic ring (SNAr) and at the sulfonyl group (SuFEx). Analysis of the reaction course revealed that deprotonation of N-nucleophile controls the reaction course: neutral amines in DMF attack at the aromatic ring, whereas amine anions, generated in equilibrium with LiHMDS in toluene, favor sulfonyl substitution. Using the base-promoted conditions we synthesized a set of substituted sulfonamides in high yields.

Ultrafine Mo2N Quantum Dots Embedded in N-Doped Graphene Sheets Enable Efficient Adsorption-Catalytic Transformation of Polysulfides.

Because of the high theoretical energy density of 2600 Wh kg-1, lithium-sulfur batteries (LSBs) are anticipated to be among the next generation of high-energy-density storage technologies. However, the practical application of LSBs has been severely hampered by the significant shuttle effect and slow redox kinetics of polysulfides (LiPSs). To address the above problems, in this paper, the concept of quantum dots (QDs) was introduced to design and synthesize Mo2N QD-modified N-doped graphene nanosheets (marked as Mo2N-QDs@NG), which were used as separator modification materials for LSBs. The experimental results demonstrated that the introduction of Mo2N QDs avoids stacking of graphene sheets and provides more active sites for the conversion of LiPSs. Moreover, Mo2N enhances the chemical fixation and catalyzes the liquid-solid conversion of soluble LiPSs by forming Mo-S and Li-N bonds with LiPSs. Additionally, establishing Mo-C bonds with Mo2N, N-doped graphene sheets can facilitate the transport of electrons and ions and physically prevent the diffusion of LiPSs, thus creating a highly conducting carbon structure to support electrochemical reactions. Benefiting from the synergistic effect of chemical immobilization and catalysis of Mo2N QDs with the physical confinement of NG, Mo2N-QDs@NG-PP batteries exhibit enhanced electrochemical performance.

Effective multi-biocatalyst system with reusable NADH for transformation of glycerol to value-added dihydroxyacetone

Glycerol-based biorefinery can be a highly profitable process by producing highly value-added products such as dihydroxyacetone via combined catalytic strategies. Here, two-enzyme system is adopted for the transformation of glycerol into highly valuable dihydroxyacetone as well as cofactor regeneration at the same time. Glycerol dehydrogenase (GDH) and alcohol dehydrogenase (ADH) are co-immobilized within magnetically separable and spherical mesocellular silica foam (Mag-S-MCF), to prepare NER-(GDH/ADH). In details, GDH and ADH are adsorbed into the mesopores of Mag-S-MCF, and further crosslinked within the mesopores of Mag-S-MCF. The resulting nanoscale enzyme reactors (NER) of crosslinked GDH and ADH molecules within the bottle-neck structured mesopores can effectively prevent larger sized crosslinked enzyme aggregates from being leached out of smaller mesopores, due to the bottle-neck mesopore structure of Mag-S-MCF, as well as stabilize the activity of GDH and ADH upon chemical crosslinking, effectively preventing the denaturation of enzyme molecules. More importantly, the proximity of GDH and ADH molecules within mesopores of NER improves the efficiency of cofactor-mediated dual-enzymatic reactions by relieving mass-transfer limitations and improving cofactor recycling in an effective way, expediting both glycerol oxidation and dihydroxyacetone generation at the same time. As a result, the DHA concentration of NER-(GDH/ADH) and the simple mixture of NER-GDH and NER-ADH were 410 μM and 336 μM, respectively. To the best of our knowledge, this report is the first demonstration of stabilized nanoscale multi-enzyme reactor system, equipped with efficient cofactor regeneration within confined mesopores, for efficient glycerol transformation to high-valued dihydroxyacetone.Graphical

The impact of digital industrialization and industrial digitalization on regional green innovation efficiency in China -From the perspective of the innovation value chain

To systematically detect the influence of digital economy on regional green innovation, this study regards regional green innovation as a two-stage sequential process, and constructs a network SBM model to evaluate the overall and stage green innovation efficiency of 30 provinces in China from 2011 to 2020. Digital economy development is decomposed into digital industrialization and industrial digitalization, and an empirical study based on fixed effects regression model of panel data is conducted to determine their influence on the overall and stage green innovation efficiency. The results suggest the following: (1) The overall efficiency in China remained at a low level, with the efficiency of research and development being slightly higher than that of achievement transformation. (2) From the national perspective, there is no discernible effect of digital industrialization on overall efficiency, but there is a negative impact on research and development and a significant positive impact on achievement transformation. Industrial digitalization significantly improves achievement transformation and overall efficiency but has no discernible effect on the research and development stage. (3) In the eastern region, industrial digitalization promotes the improvement of both stages and overall efficiency, while digital industrialization does not have a significant impact. In the central region, digital industrialization and industrial digitalization have significant positive impacts on both overall efficiency and the efficiency of achievement transformation but have no impact on research and development. In the western region, digital industrialization inhibits the overall and achievement transformation efficiency, and industrial digitalization does not have significant impact on the overall efficiency or the efficiency of each stage. Finally, some relevant policy recommendations are proposed.

CsPbX3 Nanocrystal Incorporating Transition Metal Cocatalyst for Photocatalytic Organic Transformation by Single Electron Transfer

ABSTRACTThe construction of composite photocatalysts is a promising strategy for improving the overall catalytic efficiency in organic photochemical transformation. In this study, we synthesize a series of inorganic metal perovskite CsPbX3 (X = Cl, Br, I) nanocrystals (NCs) with excellent chemical stability and wide‐range visible‐light absorption. The Pd cocatalyst is then induced to the surfaces of CsPbX3 fabricating Pd/CsPbX3 composites via visible‐light reduction method. Photoelectrochemical measurements show the introduction of Pd cocatalyst promotes efficiently the charge separation and transfer of CsPbX3. The prepared Pd/CsPbX3 composites exhibit remarkable photocatalytic efficiency in visible‐light‐induced C–C cross‐coupling reaction. The reaction is characterized by mild reaction conditions, good recyclability, and high yield from a broad variety of substrates. Mechanism investigations show the reaction relies on the synergy of CsPbX3 photocatalysis and Pd catalysis. The valence band of CsPbX3 oxidizes the phenylboronic acid into active radical species via single electron transfer. On another hand, Pd(0) as active sites occurs oxidative addition with aryl halides for achieving the C–C cross‐coupling reaction. This study displays a feasible approach for the introduction of Pd catalyst in CsPbX3 to achieve the photocatalytic organic transformation via photoinduced electron transfer.

Direct-through channels of ZIF-8 composite membranes via in-situ PolyBMA sealing: Radical-induced phase transformation and dye removal efficiency

Metal-organic frameworks (MOFs)-based hybrid composite membranes are promising candidates for achieving highly efficient water purification. In this study, a facile, novel pressure-induced filtration method involving in-situ polymerization of butyl methacrylate (BMA) was proposed for sealing ZIF-8 nanoparticles on top of polyethersulfone (PES) ultrafiltration membrane support. The developed (ZIF-8+polyBMA)/PES composite membranes with thin toplayers of <1 μm present attractive water permeance (up to 31 LMH/bar) and good rejection of Congo red (99.2 %) and Methylene blue (99.1 %). The ZIF-8 particles served as direct-through channels with superior separation performance due to their high porosity and tunable pore structure, while the interstices were successfully sealed with polyBMA by in-situ polymerization, as demonstrated by SEM and dye removal experiments. It was found that the morphology of ZIF-8 transformed from a rhombic dodecahedron to two-dimensional hexagonal nanosheets during the fabrication process. We hypothesize that this surprising phenomenon is caused by the reaction of the initiators (Na2S2O5 and K2S2O8) that generate the hydroxyl and sulfate radicals attacking and transforming the Zn-MIM bond network of ZIF-8. However, insights into the structural changes need to be further investigated to understand the mechanism involved.

Efficient genetic transformation and gene editing of Chinese cabbage using Agrobacterium rhizogenes.

A method using Agrobacterium rhizogenes-mediated callus production and plant regeneration enables efficient genetic transformation and gene editing in Chinese cabbage.

Crystal Form-Dependent MnS for Diabetic Wound Healing: Performance and Mechanistic Insights.

In pharmaceuticals, the structural and functional alterations induced by biotransformation are well-documented. Many pharmaceuticals exist in various crystal forms, which govern their transformation and significantly impact their activity. However, in the field of inorganic nanomedicine, there is a paucity of research focusing on the influence of crystal form-dependent "metabolism" (transformation) on their activity and biomechanism. This study delves into the distinct performances of two crystal forms of manganese sulfide (MnS), namely α-MnS and γ-MnS, in bacteria-infected diabetic wound healing. In the initial stage of a wound, where the environment is neutral to slightly alkaline, MnS partially converts to MnxOy (comprising Mn2O3 and Mn3O4) and concurrently produces hydrogen sulfide (H2S); the conversion efficiency of γ-MnS significantly surpasses that of α-MnS. Additionally, γ-MnS is more soluble than α-MnS, which allows it to generate more Mn2+. These components collectively contribute to the superior bacteriostatic properties of MnS. In wound related cells, MnS stimulates the production of collagen I and vascular endothelial growth factor (VEGF), promote the M1 macrophages polarizing to the M2 phenotype, for extracellular matrix (ECM) remodeling. Notably, different transformation products have distinct functions. Consequently, the activity of MnS is dependent on its original crystal form related solubility and transformation efficiency.

CRISPR-Cas9-Mediated Reduced Expression of Potato Apoplastic Invertase Inhibitor Gene and Analysis of Transformation Efficiency Parameters

CRISPR-Cas9-Mediated Reduced Expression of Potato Apoplastic Invertase Inhibitor Gene and Analysis of Transformation Efficiency Parameters