Construction and characterization of Aeromonas hydrophila crp and fur deletion mutants and evaluation of its potential as live-attenuated vaccines in crucian carp.

Aeromonas hydrophila (A. hydrophila) is a typical zoonotic pathogenic bacterium that infects humans, animals, and fish. It has been reported that the Fur, a Fe2+ regulatory protein, and the Crp, a cAMP receptor protein, play important roles in bacterial virulence in many bacteria, but no research has been investigated on A. hydrophila. In this study, the Δfur and Δcrp mutant strains were constructed by the suicide plasmid method. These two mutant strains exhibited a slightly diminished bacterial growth and also were observed some alterations in the number of outer membrane proteins, and the disappearance of hemolysis in the Δcrp strain. Animal experiments of crucian carp showed that the Δfur and Δcrp mutant strains significantly decreased virulence compared to the wild-type strain, and both mutant strains were able to induce good immune responses by two kinds of administration routes of intraperitoneal immunization (i.p) and immersion immunization, and the protection rates through intraperitoneal injection of Δfur and Δcrp to crucian carp were as high as 83.3 % and 73.3 %, respectively, and immersion immunization route of Δfur and Δcrp to crucian carp provided protection as high as 40 % and 20 %, respectively. These two mutant strains showed abilities to induce changes in enzymatic activities of the non-specific enzymes SOD, LZM, AKP, and ACP in crucian carp. Together, these results indicated the Δfur and Δcrp mutants were safe and effective candidate vaccine strains, showing good protection against the wild-type A. hydrophila challenge.

Discovery of a Novel and Potent LCK Inhibitor for Leukemia Treatment via Deep Learning and Molecular Docking.

The lymphocyte-specific protein tyrosine kinase (LCK) plays a crucial role in both T-cell development and activation. Dysregulation of LCK signaling has been demonstrated to drive the oncogenesis of T-cell acute lymphoblastic leukemia (T-ALL), thus providing a therapeutic target for leukemia treatment. In this study, we introduced a sophisticated virtual screening strategy combined with biological evaluations to discover potent LCK inhibitors. Our initial approach involved utilizing the PLANET algorithm to assess and contrast various scoring methodologies suitable for LCK inhibitor screening. After effectively evaluating PLANET, we progressed to devise a virtual screening workflow that synergistically combines the strengths of PLANET with the capabilities of Schrödinger's suite. This integrative strategy led to the efficient identification of four potential LCK inhibitors. Among them, compound 1232030-35-1 stood out as the most promising candidate with an IC50 of 0.43 nM. Further in vitro bioassays revealed that 1232030-35-1 exhibited robust antiproliferative effects on T-ALL cells, which was attributed to its ability to suppress the phosphorylations of key molecules in the LCK signaling pathway. More importantly, 1232030-35-1 treatment demonstrated profound in vivo antileukemia efficacy in a human T-ALL xenograft model. In addition, complementary molecular dynamics simulations provided deeper insight into the binding kinetics between 1232030-35-1 and LCK, highlighting the formation of a hydrogen bond with Met319. Collectively, our study established a robust and effective screening strategy that integrates AI-driven and conventional methodologies for the identification of LCK inhibitors, positioning 1232030-35-1 as a highly promising and novel drug-like candidate for potential applications in treating T-ALL.

Discovery and characterization of novel FGFR1 inhibitors in triple-negative breast cancer via hybrid virtual screening and molecular dynamics simulations.

The overexpression of FGFR1 is thought to significantly contribute to the progression of triple-negative breast cancer (TNBC), impacting aspects such as tumorigenesis, growth, metastasis, and drug resistance. Consequently, the pursuit of effective inhibitors for FGFR1 is a key area of research interest. In response to this need, our study developed a hybrid virtual screening method. Utilizing KarmaDock, an innovative algorithm that blends deep learning with molecular docking, alongside Schrödinger's Residue Scanning. This strategy led us to identify compound 6, which demonstrated promising FGFR1 inhibitory activity, evidenced by an IC50 value of approximately 0.24 nM in the HTRF bioassay. Further evaluation revealed that this compound also inhibits the FGFR1 V561M variant with an IC50 value around 1.24 nM. Our subsequent investigations demonstrate that Compound 6 robustly suppresses the migration and invasion capacities of TNBC cell lines, through the downregulation of p-FGFR1 and modulation of EMT markers, highlighting its promise as a potent anti-metastatic therapeutic agent. Additionally, our use of molecular dynamics simulations provided a deeper understanding of the compound's specific binding interactions with FGFR1.

Discovery of potent CSK inhibitors through integrated virtual screening and molecular dynamic simulation.

Oncogenic overexpression or activation of C-terminal Src kinase (CSK) has been shown to play an important role in triple-negative breast cancer (TNBC) progression, including tumor initiation, growth, metastasis, drug resistance. This revelation has pivoted the focus toward CSK as a potential target for novel treatments. However, until now, there are few inhibitors designed to target the CSK protein. Responding to this, our research has implemented a comprehensive virtual screening protocol. By integrating energy-based screening methods with AI-driven scoring functions, such as Attentive FP, and employing rigorous rescoring methods like Glide docking and molecular mechanics generalized Born surface area (MM/GBSA), we have systematically sought out inhibitors of CSK. This approach led to the discovery of a compound with a potent CSK inhibitory activity, reflected by an IC50 value of 1.6 nM under a homogeneous time-resolved fluorescence (HTRF) bioassay. Subsequently, molecule 2 exhibits strong growth inhibition of MD anderson - metastatic breast (MDA-MB) -231, Hs578T, and SUM159 cells, showing a level of growth inhibition comparable to that observed with dasatinib. Treatment with molecule 2 also induced significant G1 phase accumulation and cell apoptosis. Furthermore, we have explored the explicit binding interactions of the compound with CSK using molecular dynamics simulations, providing valuable insights into its mechanism of action.

Phase Change Microcapsules with a Polystyrene/Boron Nitride Nanosheet Hybrid Shell for Enhanced Thermal Management of Electronics.

Organic shell material and phase change material (PCM) have low thermal conductivity, which reduces the heat absorption and release rate of microencapsulated phase change materials (MEPCMs). Boron nitride nanosheets (BNNSs) with high thermal conductivity can not only stabilize the oil phase as the Pickering emulsifier but also improve the thermal conductivity of MEPCMs as one of the shell components, thus facilitating the heat conduction in the microcapsule system. Herein, MEPCM with paraffin wax (PW) as the core material and polystyrene (PS) modified by BNNSs as the shell material (PW@PS/BNNS MEPCMs) are synthesized via Pickering emulsion polymerization. The structure of PW@PS/BNNS MEPCMs can be regulated by tuning the PW and BNNS contents, to achieve high latent heat and thermal conductivity. In comparison to pure PW, the thermal conductivity of MEPCMs-5 wt % BNNSs increases by 63.76% at 25 °C. The PW@PS/BNNS powder possesses a latent heat capacity of 166.3 J/g, corresponding to a high encapsulation ratio of 80.77%. These properties endow the prepared MEPCMs with excellent thermal regulation properties. We also propose the formation mechanism of PW@PS/BNNS MEPCMs via Pickering emulsion polymerization for the first time, which will guide the MEPCM fabrication toward a reliable direction.

Effective Structure Control of Colloidal Molecules and the Morphology Evolution Mechanism Investigation.

Colloidal molecules (CMs), nonspherical clusters of a small number of particles, can be used as building blocks for self-assembly applications. Here, we propose a novel one pot method for CMs synthesis. First, poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) microgels were prepared by soap-free emulsion polymerization as seed particles, then monomer styrene and cross-linking agent divinylbenzene (DVB) were added, which could be polymerized by the remaining free radicals on the seed surface in situ. P(NIPAM-co-AA)-PS colloidal molecules with a series of morphologies such as popcorn-like, CO2-like, NH3-like, CH4-like and so on could be obtained. The effects of satellite colloid viscosity, interfacial tension, and polymer chain mobility on the number of satellite colloid have been investigated, and the formation mechanism of CMs is proposed based on morphology evolution investigation. Compared with the existing CM synthesis techniques, our method enables fabricating CMs from vinyl monomer in a facile and efficient way, and the scientific finding regarding the CMs formation will guide the CMs fabrication toward salable and reliable direction.

Preparation of carbon quantum dots based high photostability luminescent membranes.

Urethane acrylate (UA) was used to prepare carbon quantum dots (C-dots) luminescent membranes and the resultants were examined with FT-IR, mechanical strength, scanning electron microscope (SEM) and quantum yields (QYs). FT-IR results showed the polyurethane acrylate (PUA) prepolymer -C = C-vibration at 1101 cm-1 disappeared but there was strong vibration at1687cm-1 which was contributed from the-C = O groups in cross-linking PUA. Mechanical strength results showed that the different quantity of C-dots loadings and UV-curing time affect the strength. SEM observations on the cross-sections of the membranes are uniform and have no structural defects, which prove that the C-dots are compatible with the water-soluble PUA resin. The C-dot loading was increased from 0 to 1 g, the maximum tensile stress was nearly 2.67 MPa, but the tensile strain was decreased from 23.4% to 15.1% and 7.2% respectively. QYs results showed that the C-dots in the membrane were stable after 120 h continuous irradiation. Therefore, the C-dots photoluminescent film is the promising material for the flexible devices in the future applications.

A Novel Piezoresistive Accelerometer with SPBs to Improve the Tradeoff between the Sensitivity and the Resonant Frequency.

For improving the tradeoff between the sensitivity and the resonant frequency of piezoresistive accelerometers, the dependency between the stress of the piezoresistor and the displacement of the structure is taken into consideration in this paper. In order to weaken the dependency, a novel structure with suspended piezoresistive beams (SPBs) is designed, and a theoretical model is established for calculating the location of SPBs, the stress of SPBs and the resonant frequency of the whole structure. Finite element method (FEM) simulations, comparative simulations and experiments are carried out to verify the good agreement with the theoretical model. It is demonstrated that increasing the sensitivity greatly without sacrificing the resonant frequency is possible in the piezoresistive accelerometer design. Therefore, the proposed structure with SPBs is potentially a novel option for improving the tradeoff between the sensitivity and the resonant frequency of piezoresistive accelerometers.

A MEMS Resonant Sensor to Measure Fluid Density and Viscosity under Flexural and Torsional Vibrating Modes.

Methods to calculate fluid density and viscosity using a micro-cantilever and based on the resonance principle were put forward. Their measuring mechanisms were analyzed and the theoretical equations to calculate the density and viscosity were deduced. The fluid-solid coupling simulations were completed for the micro-cantilevers with different shapes. The sensing chips with micro-cantilevers were designed based on the simulation results and fabricated using the micro electromechanical systems (MEMS) technology. Finally, the MEMS resonant sensor was packaged with the sensing chip to measure the densities and viscosities of eight different fluids under the flexural and torsional vibrating modes separately. The relative errors of the measured densities from 600 kg/m³ to 900 kg/m³ and viscosities from 200 µPa·s to 1000 µPa·s were calculated and analyzed with different microcantilevers under various vibrating modes. The experimental results showed that the effects of the shape and vibrating mode of micro-cantilever on the measurement accuracies of fluid density and viscosity were analyzed in detail.

A Novel Slope Method for Measurement of Fluid Density with a Micro-cantilever under Flexural and Torsional Vibrations.

A novel method, which was called a slope method, has been proposed to measure fluid density by the micro-cantilever sensing chip. The theoretical formulas of the slope method were discussed and established when the micro-cantilever sensing chip was under flexural and torsional vibrations. The slope was calculated based on the fitted curve between the excitation and output voltages of sensing chip under the nonresonant status. This measuring method need not sweep frequency to find the accurate resonant frequency. Therefore, the fluid density was measured easily based on the calculated slope. In addition, the micro-cantilver was drived by double sided excitation and free end excitation to oscillate under flexural and torsional vibrations, respectively. The corresponding experiments were carried out to measure the fluid density by the slope method. The measurement results were also analyzed when the sensing chip was under flexural and torsional nonresonant vibrations separately. The measurement accuracies under these vibrations were all better than 1.5%, and the density measuring sensitivity under torsional nonresonant vibration was about two times higher than that under flexural nonresonant vibration.

DRIFTS evidence for facet-dependent adsorption of gaseous toluene on TiO2 with relative photocatalytic properties.

Effective adsorption is of great importance to the photocatalytic degradation of volatile organic compounds. Herein, we succeeded in the preparation of anatase TiO2 with clean dominant {001} and {101} facets. By using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) equipped with a homemade reaction system and a coupling gas-dosing system, we found that TiO2 with dominant {001} facets exhibits higher toluene adsorption capacity than TiO2 with dominant {101} facets, which may be attributed to the different number of unsaturated 5c-Ti capable of forming the main active adsorption sites (terminal Ti-OH species). TiO2 with dominant {001} facets shows a significantly high photocatalytic degradation performance, with its degradation rate being 6 times higher than that of dominant {101} facets. Combined with simulation results, it is suggested that the synergetic effects of the formation of specific active adsorption sites, the low adsorption energy for toluene, and preservation of the free molecularly adsorbed water on the surface promote the degradation of gaseous toluene on the dominant {001} facets. This study exemplifies that the facet-dependent adsorption of volatile organic compounds is one of the most important factors to effectively engineer photocatalysts for air purification.

Corrigendum: The influence of physical education courses integrated with civic education on prosocial behavior among college students: the chain mediating effect of cultural confidence and self-esteem.

[This corrects the article DOI: 10.3389/fpsyg.2023.1217290.].

Construction and Evaluation of an Efficient Live Attenuated Salmonella Choleraesuis Vaccine and Its Ability as a Vaccine Carrier to Deliver Heterologous Antigens.

The gram-negative facultative intracellular pathogen Salmonella enterica serotype Choleraesuis, also known as S. Choleraesuis, is a major financial loss for the pig business. C500 is a vaccine strain that has been used for preventing S. Choleraesuis infection in pigs for many years in China. Although it possessed good immunogenicity and protection efficacy, it still showed severe side effects. The truncation of the key gene rpoS in C500 was believed to take the major responsibility for its attenuation. To achieve a good balance between attenuation and immunogenicity, rpoS was restored to an active state, and other essential virulent genes of crp, fur, phoP, and aroA were evaluated for their effects of deletion on safety and immunogenicity. Animal experiments demonstrated that C5001 (C500 rpoS+ Δcrp10) and C5002 (C500 rpoS+ Δfur9) showed an excellent ability to induce an immune response. To further decrease the endotoxic activity, the combination mutations of ΔpagL7 ΔpagP81::PlpplpxE ΔlpxR9 were introduced into the mutant strains to generate 1'-dephosphorylated lipid A. Animal experiments showed that SC3 (C500 rpoS+ Δfur9 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) induced higher levels of IgG and secreted IgA antibodies and provided a higher protection rate than SC1 (C500 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) and SC2 (C500 rpoS+ Δcrp10 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9). We also evaluated the ability of SC3 (C500 rpoS+ Δfur9 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) as a vaccine carrier to deliver heterologous protein antigens and polysaccharide antigens. The results indicated that SC3 (C500 rpoS+ Δfur9 ΔpagL7 ΔpagP81:: PlpplpxE ΔlpxR9) showed an excellent ability to deliver heterologous antigens and induce the host to produce high levels of antibodies. Together, these results indicate that we constructed a safe and efficient attenuated strain of the S. Choleraesuis vaccine, which demonstrated strong resistance to infection with wild-type S. Choleraesuis and can be employed as a universal vector for the delivery of recombinant antigens.

Icephobic Durability of Molecular Brush-Structured PDMS Soft Coatings.

The accumulation of ice can pose numerous inconveniences and potential hazards, profoundly affecting both human productivity and daily life. To combat the challenges posed by icing, extensive research efforts have been dedicated to the development of low-ice adhesion surfaces. In this study, we harness the power of molecular dynamics simulations to delve into the intricate dynamics of polymer chains and their role in determining the modulus of the material. We present a novel strategy to prepare ultralow-modulus poly(dimethylsiloxane) (PDMS) elastomers with a molecular brush configuration as icephobic materials. The process involves grafting monohydride-terminated PDMS (H-PDMS) as side chains onto backbone chain PDMS with pendant vinyl functional groups to yield a molecular brush structure. The segments of this polymer structure effectively restrict interchain entanglement, thereby rendering a lower modulus compared to traditional linear structures at an equivalent cross-linking density. The developed soft coating exhibits a remarkably ultralow ice adhesion strength of 13.1 ± 1.1 kPa. Even after enduring 50 cycles of icing and deicing, the ice adhesion strength of this coating steadfastly stayed below 16 kPa, showing no notable increase. Importantly, the molecular brush coating applied to glass demonstrated an impressive light transmittance of 92.1% within the visible light spectrum, surpassing the transmittance of bare glass, which was measured at 91.3%. This icephobic coating with exceptional light transmittance offers a wide range of applications and holds significant potential as a practical icephobic material.

Discovery of anti-melanogenic components in persimmon (Diospyros kaki) leaf using LC-MS/MS-MN, AlphaFold2-enabled virtual screening and biological validation.

Persimmon (Diospyros kaki) leaf is widely used as a tea substitute in East Asia, offering potential health benefits. Although studies have highlighted their effects on hyperpigmentation disorders, the active components remain unidentified. This study introduces a novel approach combining LC-MS/MS-based molecular networking with AlphaFold2-enabled virtual screening to expedite the identification of bioactive components in persimmon leaf. A total of 105 compounds were identified by MS/MS analysis. Further, virtual screening identified five flavonoids with potential anti-melanogenic properties. Bioassays confirmed myricetin, quercetin, and kaempferol inhibited melanogenesis in human melanocytes in a dose-dependent manner. Biolayer interferometry assays revealed strong binding affinity between these flavonols and hsTYR, with KD values of 23.26 ± 11.77 for myricetin, 12.43 ± 0.37 for quercetin, and 14.99 ± 3.80 µM for kaempferol. Molecular dynamics simulations provided insights into the binding interactions of these flavonols with hsTYR, particularly highlighting the essential role of the 3-OH group on the C-ring. This study elucidates the bioactive components responsible for the anti-melanogenic effects of persimmon leaf, supporting their use in product development.

Research on point cloud hole filling and 3D reconstruction in reflective area.

3D reconstruction is the process of obtaining the three-dimensional shape or surface structure of an object, which is widely used in advanced manufacturing fields such as automotive, aerospace, industrial inspection, and reverse engineering. However, due to the structural characteristics of the component itself, the reflective properties of the coating material, and other factors, there may be specular reflection during image acquisition, making it difficult to achieve complete 3D reconstruction of the component. This paper proposes a method to address the problem of incomplete 3D reconstruction of strongly reflective objects by recognizing outlier points and filling point cloud holes. The proposed View-Transform-PointNet outlier point recognition network improves the alignment of the initial point cloud plane and implements secondary alignment of the point cloud based on the perpendicularity between the outlier plane in mixed reflection and the point cloud plane. The point cloud hole-filling method is based on the principle of outlier formation and approximates a local Gaussian distribution to linear variation. The distance between the end of each outlier plane and the real surface is calculated to repair the depth information of outlier points. The proposed method achieves a 39.4% increase in the number of point cloud filling, a 45.2% increase in the number of triangular mesh faces, a 46.9% increase in surface area, and a chamfer distance (CD) of 0.4471009, which is better than existing geometric repair methods in terms of standard deviation and smoothness. The method improves the alignment of initial point cloud planes and enhances the accuracy of outlier point recognition, which are the main innovative points of this study. The 3D reconstruction of the repaired point cloud model is achieved through Poisson equation and parameter adjustment. The proposed method reduces the error caused by large curvature in the boundary region and improves the smoothness and accuracy of the reconstructed model.

Surface-enhanced Raman scattering of a gold core-silver shell-sponge substrate for detection of thiram and diquat.

Aiming at the difficulty of traditional pesticide sampling, a low-cost and convenient flexible surface enhanced Raman scattering (SERS) gold core-silver shell-sponge (Au-Ag-sponge) substrate was synthesized by chemical reduction. The SERS substrate consisted of Au-AgNPs and a melamine sponge. The sponge had a rich open pore structure, which could well "capture" Au-AgNPs, generating a large number of "hot spots". The SERS enhancement activity of the flexible substrate was characterized with rhodamine 6G (R6G) Raman probe molecules. The substrate showed good activity to 10-12 M rhodamine 6G with an enhancement factor (EF) of 7.72 × 106. Applying this substrate to the qualitative and quantitative detection of pesticide residues, the results showed that the Raman intensity was well related to the concentration of pesticide solution with the range of 0.1-10 mg L-1 of thiram and 1-10 mg L-1 of diquat. Furthermore, the substrate was analyzed by finite difference time domain (FDTD) simulation and the results were in good agreement with the experimental results. The reason for the difference in Raman signals of pesticide molecules on the same substrate was the different binding modes of Au-AgNPs on the sponge. Finally, we pointed out the advantages of flexible substrates in the field of pesticide residues, as well as future opportunities and challenges.

Research on a three-dimensional SERS substrate based on a CNTs/Ag@Au/SiO2 composite for detection of fipronil and imidacloprid pesticides.

Surface-enhanced Raman scattering (SERS) has a unique fingerprint spectrum, which allows for rapid, highly sensitive, and non-destructive detection without the need for sample pretreatment. However, SERS substrates have disadvantages such as short storage time and poor reproducibility. In this study, carbon nanotubes, gold, and silver were combined to take advantage of their inherent structural and characteristic properties that enhance the Raman effect. A new type of SERS composite substrate, CNTs/Ag@Au/SiO2, was prepared using a hydrothermal method and seed growth method. The substrate was characterized using transmission electron microscopy (TEM), and the average distance between the core-shell nanoparticles was found to be 3.1 nm, which is more suitable than other gold-silver combined core-shell structures and significantly improves the SERS enhancement factor. The substrate demonstrated high sensitivity even at low concentrations of probe molecules and good uniformity at five randomly selected locations. After storage for 45 days, the substrate still exhibited good stability. In most gold-silver combined core-shell structures, the detection limit for Rhodamine 6G (R6G) is 10-9 mol L-1, while in this substrate, the detection limit for R6G is 10-11 mol L-1. Furthermore, the contribution of the substrate's enhancement was deeply investigated using finite-difference time-domain (FDTD), which revealed that the substrate's hotspots were present in two forms: the "hotspots generated between Ag@Au nanoparticles" and the "hotspots generated between Ag@Au nanoparticles and carbon nanotubes". These two forms of hotspots also demonstrated that the performance brought about by the preparation of the substrate structure was reliable. The simulation results were compared with the experimental results, and the analysis showed that the real environment would have an impact on the substrate's structure during the actual substrate preparation process. Finally, the substrate was used for detecting the pesticide fipronil, and the results showed clear peaks even at a concentration of 0.1 mg L-1. The results indicated that the Raman intensity was linearly exponential with the fipronil solution concentration, with a determination coefficient of R2 = 0.991. This study provides a new SERS substrate for pesticide residue detection and further explores the improvement of pesticide detection limits.

Single-molecule lipopolysaccharides identification and the interplay with biomolecules via nanopore readout.

Lipopolysaccharides (LPS) are the major constituent on the cell envelope of all gram-negative bacteria. They are ubiquitous in air, and are toxic inflammatory stimulators for urinary disorders and sepsis. The reported optical, thermal, and electrochemical sensors via the intermolecular interplay of LPS with proteins and aptamers are generally complicated methods. We demonstrate the single-molecule nanopore approach for LPS identification in distinct bacteria as well as the serotypes discrimination. With a 4 nm nanopore, we achieve a detection limit of 10 ng/mL. Both the antibiotic polymyxin B (PMB) and DNA aptamer display specific binding to LPS. The identification of LPS in both human serum and tap water show good performance with nanopore platforms. Our work shows a highly-sensitive and easy-to-handle scheme for clinical and environmental biomarkers determination and provides a promising screening tool for early warning of contamination in water and medical supplies.

Novel biodegradation pathway of insecticide flonicamid mediated by an amidase and its unusual substrate spectrum.

The insecticide flonicamid (FLO) and its main degradation intermediate 4-trifluoromethylnicotinamide (TFNA-AM) are hazardous to the environment and animals. Microbial transformation of FLO has been well studied, but no study has yet reported on TFNA-AM degradation by a microorganism. Here, Pseudomonas stutzeri CGMCC 22915 effectively degraded TFNA-AM to 5-trifluoromethylnicotinic acid (TFNA). P. stutzeri CGMCC 22915 degraded 60.0% of TFNA-AM (1154.44 µmol/L) within 6 h with a half-life of just 4.5 h. Moreover, P. stutzeri CGMCC 22915 significantly promoted TFNA-AM decomposition in surface water. The reaction was catalyzed by an amidase, PsAmiA. PsAmiA is encoded in a novel nitrile-converting enzyme gene cluster. The enzyme shared only 20-44% identities with previously characterized signature amidases. PsAmiA was successfully expressed in Escherichia coli and its enzymatic properties were investigated using TFNA-AM as the substrate. PsAmiA was more active toward amides without hydrophilic groups, and did not hydrolyze another amide metabolite of FLO, N-(4-trifluoromethylnicotinoyl)glycinamide (TFNG-AM), which is structurally very similar to TFNA-AM. Molecular docking of PsAmiA and TFNA-AM indicated that hydrophobic residues Leu148, Ala150, Ala195, Ile225, Trp341, Leu460, and Ile463 may affect its substrate spectrum. This study provides new insights of the environmental fate of FLO at the molecular level and the structure-function relationships of amidases.