Circular Dichroism Analysis Research Articles

Alkaloids with Their Protective Effects Against Aβ25-35-Induced PC-12 Cell Injury from the Tubers of Pinellia pedatisecta Schott

Seven new alkaloids [1, (±)-2, (±)-3, 4, and 5] and one new natural product (6), along with eight known analogues, were isolated from the tubers of Pinellia pedatisecta Schott. Their structures were determined by a comprehensive analysis of spectroscopic data, including HRESIMS, and electronic circular dichroism (ECD). In addition, the results of the bioactivity evaluation showed that compounds (±)-3, 6, and 9 exhibited significantly protective effects against Aβ25-35-induced PC-12 cell injury and ameliorated cell viabilities by decreasing the levels of the reactive oxidative species (ROS) and mitochondrial membrane potential (MMP).

Simple solution and paper-based fluorescent aptasensors for toxic metal ions, thallium(l) and lead(ll).

Heavy metal ions, such as thallium(I) and lead(II) are environmental toxicants known to cause a severe threat to human and ecosystem health. This work investigates aptamers and intercalating chromophore-based complexes for the detection of these toxic species. In one method, a selective label-free "turn-on" biosensor was developed using a G-quadruplex-intercalating agent, berberine. Fluorescence, melting temperature (Tm), and circular dichroism analysis confirmed the affinity and selectivity results, illustrating the potential of these aptasensor methods for improving detection limits. These fluorescence assays were found to perform with a detection limit of 3.4 μM for Tl(I) and 0.84 nM for Pb(II). Furthermore, the assays were challenged successfully with Tl(I) and Pb(II) spiked into river water samples. We next developed paper-based fluorescent assays for Tl(I) and Pb(II), where the aptamer/berberine complex was spotted onto the paper test zone. When Tl(I) or Pb(II) ions solutions were spotted onto the top of the test zone and the spot was illuminated with a portable UV light (365 nm), a strong green fluorescence could be easily visualized with the naked eye. The lowest detection limits achieved with these fluorescent paper-based assays for Tl(I) and Pb(II) were 1.1 nM and 1.6 nM, respectively. The two fluorescent approaches presented here have the potential to be the basis of rapid, fast, and cost-efficient screening assays for these toxic species.

Modifications in Protein Structure and Xanthine Oxidase Inhibition of Yak Casein Induced by Protease Treatment

ABSTRACTYak milk is abundant in casein, which can generate a variety of bioactive peptides through enzymatic hydrolysis. However, the influence of enzymatic hydrolysis on the structural properties of yak casein and its inhibitory effects on xanthine oxidase (XOD) remain largely unexplored. This study demonstrated that when yak casein was subjected to treatment with flavourzyme, the degree of hydrolysis progressively increased over time, resulting in the fragmentation of the casein's flake‐like structure into smaller particles. Circular dichroism analysis revealed that after 4 h of enzymatic treatment, there was an elevation in the β‐sheet content of the yak casein hydrolysate, while other secondary structure elements diminished. Furthermore, flavourzyme treatment induced modifications in the tertiary structure of yak casein. The study also examined the impact of varying hydrolysis durations on XOD inhibitory activity, discovering that the hydrolysate obtained after 3 h displayed the highest inhibition on XOD, with an inhibition rate of (40.63 ± 3.36) %. Additionally, the fraction of the hydrolysate with a molecular weight exceeding 3 kDa demonstrated enhanced XOD inhibitory activity. This study is the first to investigate how varying hydrolysis durations with flavourzyme affect the structural characteristics of yak casein and its XOD inhibitory activity.

Triantaspirols A-C and Paraphaeolactone Cs from Paraphaeosphaeria sp. KT4192: Sensitivity of CP3 in Distinguishing Close NMR Signals.

Hybridized spirobisnaphthalene derivatives, triantaspirols A-C (1-3) and paraphaeolactones C1 and C2 (4 and 5), were identified from the culture broth of the fungus Paraphaeosphaeria sp. KT4192. The NMR spectra of 2 and 3, as well as 4 and 5, closely resembled each other, indicating that these were pairs of diastereomers. Although this NMR spectral resemblance made it challenging to distinguish their relative configurations, detailed analysis of the electronic circular dichroism (ECD) spectra and NOE correlations allowed us to deduce them. The CP3 metric with the DFT-based NMR chemical shifts was found to distinguish configurations of diastereomers in a highly sensitive and accurate manner that DP4 could not account for because of the very close chemical shift differences in the experimental NMR spectra. The reliability of this method was assessed using 23 published examples which could not be distinguished by DP4 protocol.

Preparation and characterization of ovomucin self-assembly nanoparticles under glycerol compression for astaxanthin delivery: Sustained release and antioxidant activity.

Astaxanthin (AST) is a natural hydrophobic nutrient with various biological activities, but its low solubility limits its application. In this study, self-assembly nanoparticles were prepared by ovomucin (OVM) and Ca2+ with the enhancement of glycerol to deliver AST. Glycerol compressed the particle size of nanoparticles from 175.7±1.8 to 142.9±0.6nm, and the nanoparticles had a strong negative charge (-28.9±0.6mV). Ultraviolet-visible spectroscopy and X-ray diffraction (XRD) confirmed the successful encapsulation of AST in an amorphous form with a high encapsulation efficiency (82.9%±2.1%). Fourier transform infrared and circular dichroism analyses demonstrated that nanoparticles formation mainly involved electrostatic interactions and hydrophobic interactions. AST in nanoparticles presented excellent gastric juice resistance and sustained release ability, whereas free radical scavenging efficiency reached up to 75%. In addition, the nanoparticles had no apparent toxicity to cell viability. This study is expected to provide a new insight into the safe and efficient delivery of AST, while demonstrating the potential of OVM as a delivery carrier in the food and health industries.

Sesquiterpenes from the leaves of Artemisia argyi and evaluation of their in vitro anti-inflammatory activities

Two unreported sesquiterpenoids, 7-en-13-hytanaphallin (1) and 8´-en-achillinin C (2) along with two known sesquiterpenoid dimers were obtained from chloroform parts in the leaves of Artemisia argyi Lévl. et Vant, belonging to Asteraceae family. The planar structures and absolute configurations of two new compounds were characterized by HR-ESI-MS, IR, 1D and 2D NMR, and electronic circular dichroism analyses. Two known sesquiterpenoids, argyinolide O (3) and (+)-8-acetylarteminolide (4), exhibited potent activities against NO production from LPS-induced RAW264.7 cells with IC50 values of 3.6 and 9.8 μM, respectively.

Exploring the salty peptides of enzymatically hydrolyzed Volvariella volvacea protein: Structural analysis and taste perception insights

This study aimed to prepare and screen salty peptides and elucidate the factors triggering their saltiness properties and saltiness-enhancing effects. Electronic tongue results indicated that samples treated with Alcalase (AJ) exhibited the highest saltiness intensity (5.05 ± 0.11) and greater saltiness-enhancing compared to those treated with Flavourzyme (AF), Neutrase (AZ), and Papain (AM). Peptides identified by LC-MS/MS were docked to the TMC4 receptor, resulting in 23 peptides with good docked results. The key amino acid residues (Thr431, Arg433, Phe440, Phe432, and Ser273) were discovered. The salty peptides WPGFK (633.75 Da), YFDWPGFK (1059.17 Da), and YFDWPGFKT (1160.27 Da) were selected for salt reduction and molecular characterization studies. Electronic tongue results showed that the 0.01 % WPGFK solution matched the saltiness of the 0.24 % NaCl solution and demonstrated superior saltiness-enhancing compared to YFDWPGFK and YFDWPGFKT. Primary sequence analysis of three salty peptides suggested that the WPG amino acid segment might play a crucial role in generating saltiness. Circular dichroism (CD) analysis of the secondary structure indicated that increased random coil and decreased β-turn contents resulted in higher saltiness perception and increased random coil content led to greater saltiness-enhancing ability.

A comprehensive investigation of the nano-[Cu2-(DIP)2-EA] effects on HSA through spectroscopic procedures and computer simulations

In this research, the toxicity of nano-[Cu2-(DIP)2-EA], a metal nano-complex consisting of ellagic acid and bathophenanthroline ligands, on human serum albumin (HSA) at a protein level was investigated. Molecular docking simulations and spectral analyses were conducted in a simulated physiological environment at pH 7.4 to explore the interaction of nano-[Cu2-(DIP)2-EA] with HSA. The results represented an increase in albumin absorption upon exposure to nano-[Cu2-(DIP)2-EA], demonstrating significant interaction between the two compounds. Steady-state and time-resolved fluorescence measurements pointed out that nano-[Cu2-(DIP)2-EA] induced static quenching of the albumin's intrinsic fluorescence with a high binding affinity of approximately 106 mol/L in a 1:1 interaction ratio. The thermodynamic variables clarified that binding of nano-[Cu2-(DIP)2-EA] to albumin occurs spontaneously and primarily driven by van der Waals interactions and H-bonds. The results of the computer simulations and the binding displacement experiments utilizing the site markers warfarin and ibuprofen revealed that nano-[Cu2-(DIP)2-EA] binds to site I within the subdomain IIA of albumin. Circular dichroism analysis elaborated that nano-[Cu2-(DIP)2-EA] slightly perturbed the microenvironment around of tryptophan residues and diminished the α-helix structure stability to a negligible amount.

Exploring the Kinetics and Thermodynamics of a Novel Histidine Ammonia-Lyase from Geobacillus kaustophilus.

Histidine ammonia-lyase (HAL) plays a pivotal role in the non-oxidative deamination of L-histidine to produce trans-urocanic, a crucial process in amino acid metabolism. This study examines the cloning, purification, and biochemical characterization of a novel HAL from Geobacillus kaustophilus (GkHAL) and eight active site mutants to assess their effects on substrate binding, catalysis, thermostability, and secondary structure. The GkHAL enzyme was successfully overexpressed and purified to homogeneity. Its primary sequence displayed 40.7% to 43.7% similarity with other known HALs and shared the same oligomeric structure in solution. Kinetic assays showed that GkHAL has optimal activity at 85 °C and pH 8.5, with high thermal stability even after preincubation at high temperatures. Mutations at Y52, H82, N194, and E411 resulted in a complete loss of catalytic activity, underscoring their essential role in enzyme function, while mutations at residues Q274, R280, and F325 did not abolish activity but did reduce catalytic efficiency. Notably, mutants R280K and F325Y displayed novel activity with L-histidinamide, expanding the substrate specificity of HAL enzymes. Circular dichroism (CD) analysis showed minor secondary structure changes in the mutants but no significant effect on global GkHAL folding. These findings suggest that GkHAL could be a promising candidate for potential biotechnological applications.

Spectroscopic Response of Chiral Proteophenes Binding to Two Chiral Insulin Amyloids

We recently reported on 8 different pentameric oligothiophenes, denoted proteophenes, with different amino acid substitution patterns along the conjugated thiophene backbone. The proteophenes were forming chiral both molecular (solvent dissolved) and self‐assembled nano‐architectonic structures, the latter having approximately 5‐6 times greater ICD responses. Herein, two proteophenes, HS‐84‐E‐E and HS‐84‐K‐K, were further investigated in terms of binding to two chiral variants of insulin amyloid fibrils. Binding curves based on fluorescence revealed strong primary binding sites for both proteophenes and more unspecific secondary binding for especially the latter. Interestingly, their induced circular dichroism (ICD) responses were similar to or stronger than for their solvent form and showed a complicated alteration upon binding suggesting that the microstructure at the binding site governs the helical structure of the ligand. Hyperspectral fluorescence microscopy and FLIM revealed more details on the molecular binding in terms of proteophene emission decay‐times. Vibrational circular dichroism (VCD) analysis showed that VCD signal of amyloid fibrils was enhanced upon binding of proteophenes, suggesting changes in the amyloid microstructures.

Selective Interaction of Chiral Carbon Dots with Nucleic Acids: A Promising Nanosensing Platform.

Carbon dots (CDs) represent an emerging class of nanomaterials that combine outstanding photoluminescent properties with low toxicity and excellent biocompatibility. These unique features have garnered significant interest for potential applications in sensing as well as nanovectors for bioactive compounds. Within this context, the possibility of synthesizing chiral carbon dots (CCDs) has paved the way for a plethora of bioapplications in their interaction with chiral biomolecules. In this study we report the synthesis and characterization of CCDs with opposite chiralities and their selective interaction with nucleic acids. A systematic study on their interaction with different oligonucleotides (ODNs) using UV-vis, photoluminescence, and circular dichroism analyses highlighted how the chiral surface of the CCDs induces distinct spectroscopic responses in CCDs-ODN conjugates. These findings establish the foundation for innovative applications of CCDs as nanosensors and nanocarriers for nucleic acids. Additionally, the antioxidant properties of CCDs were investigated, highlighting their dual potential as both sensing and preservative nanomaterials for genetic material. Our results suggest significant implications for the development of chiral-specific diagnostic tools, drug delivery systems, and therapeutic agents. Furthermore, these properties open new avenues for the use of CCDs in antibiotic residue detection, fluorescence imaging, and photodynamic therapy.

New Sesquiterpenoids from the Mangrove-Derived Fungus Talaromyces sp. as Modulators of Nuclear Receptors.

Four new sesquiterpenoids, talaroterpenes A-D (1-4), were isolated from the mangrove-derived fungus Talaromyces sp. SCSIO 41412. The structures of compounds 1-4 were elucidated through comprehensive NMR and MS spectroscopic analyses. The absolute configurations of 1-4 were assigned based on single-crystal X-ray diffraction and calculated electronic circular dichroism analysis. Talaroterpenes A-D (1-4) were evaluated with their regulatory activities on nuclear receptors in HepG2 cells. Under the concentrations of 200 μM, 1, 3 and 4 exhibited varying degrees of activation on ABCA1 and PPARα, while 4 showed the strongest activities. Furthermore, 4 induced significant alterations in the expression of downstream target genes CLOCK and BMAL1 of RORα, and the in silico molecular docking analysis supported the direct binding interactions of 4 with RORα protein. This study revealed that talaroterpene D (4) was a new potential non-toxic modulator of nuclear receptors.

Association mechanism of bicalutamide and human serum albumin for potential clinical implications.

The binding mechanism of molecular interaction between bicalutamide and human serum albumin (HSA) in a pH7.4 phosphate buffer was studied using various spectroscopic techniques in combination with molecular modeling. Fluorescence data revealed that the fluorescence quenching of HSA by bicalutamide was a static quenching procedure. The binding constants and number of binding sites were evaluated at different temperatures. The thermodynamic parameters, ΔH and ΔS, were calculated to be 4.30 × 104J·mol-1 and 245 J·mol-1·K-1, respectively, suggesting that the binding of bicalutamide to HSA was driven mainly by hydrophobic interactions and hydrogen bonds. The displacement studies indicated neither Sudlow's site I nor II but subdomain IB as the main binding site for bicalutamide on HSA. The binding distance between bicalutamide and HSA was determined to be 3.54 nm based on the Förster theory. Analysis of circular dichroism, synchronous, and 3D fluorescence spectra demonstrated that HSA conformation was slightly altered in the presence of bicalutamide.

Discovery of antitumor diterpenoids from Casearia graveolens targeting VEGFR-2 to inhibit angiogenesis

Eight novel clerodane diterpenoids (1−8) were isolated from the twigs of Casearia graveolens. Their structures were elucidated through comprehensive nuclear magnetic resonance (NMR), high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), and electronic circular dichroism (ECD) analyses. In addition to structural determination, surface plasmon resonance (SPR) assays were conducted to investigate molecular interactions, revealing that compound 8 exhibited high affinity for vascular endothelial growth factor receptor 2 (VEGFR2), a key regulator of tumor angiogenesis. Subsequent in vivo experiments demonstrated that compound 8 effectively inhibited angiogenesis and displayed significant antitumor activity by suppressing tumor proliferation and metastasis in zebrafish xenograft models. These findings suggest that compound 8 holds promise as an anticancer lead compound targeting VEGFR-2 to obstruct tumor angiogenesis.

Comparative Study on the Postmortem Proteolysis and Shear Force during Aging of Pork and Beef Semitendinosus Muscles.

The differences in the proteolytic patterns and shear force of pork and beef during aging were evaluated. Pork and beef semitendinosus muscles were obtained at 24 and 48 h postmortem, respectively, and aged at 4°C for 0 (Day 0), 7 (Day 7), and 14 days (Day 14). Changes in the electrical conductivity were observed in pork on Day 7 and beef on Day 14. The calpain activity increased in pork (p<0.05) after 14 days of aging, whereas that of beef decreased on Day 7 (p<0.05). The cathepsin B activity in pork and beef increased between Day 7 and 14 (p<0.05). The content of α-amino group in the 10% trichloroacetic acid-soluble fraction increased between Day 7 and 14 in pork (p<0.05), but increased steadily in beef throughout aging (p<0.05). The electrophoretogram of the myofibrillar proteins revealed a 30 kDa protein band only in the beef lane on Day 14. The cooked pork had no significant changes in the shear force during aging periods (p>0.05), while the gradual decrease in the shear force with the increasing aging periods was shown in the cooked beef (p<0.05). Circular dichroism analysis of myosin extracts from pork and beef revealed thermal denaturation temperatures of 55°C and 58°C, respectively. This study highlights the different post-mortem proteolytic patterns and thermal denaturation temperatures of myosin in pork and beef semitendinosus muscles, which contribute to distinct changes in the shear force during aging between pork and beef.

Evaluating the Antimicrobial Efficacy of a Designed Synthetic Peptide against Pathogenic Bacteria.

Recent research has focused on discovering peptides that effectively target multi-resistant bacteria while leaving healthy cells unharmed. In this work, we describe the antimicrobial properties of RK8, a peptide composed of eight amino acid residues. Its activity was tested against multidrug-resistant Gram-negative and Gram-positive bacteria. RK8's efficacy in eradicating mature biofilm and increasing membrane permeability was assessed using Sytox Green. Cytotoxicity assays were conducted both in vitro and in vivo models. Circular dichroism analysis revealed that RK8 adopted an extended structure in water and sodium dodecyl sulfate (SDS). RK8 exhibited MICs of 8-64 μM and MBCs of 4-64 μM against various bacteria, with higher effectiveness observed in Staphylococcus aureus (MRSA) and E. coli KPC+ strains than others. Ciprofloxacin and Vancomycin showed varying MIC and MBC values lower than RK8 for Gram-positive bacteria, but competitive for Gram-negative bacteria. The combination of RK8 and ciprofloxacin showed a synergistic effect. The RK8 peptides could reduce 38% of the mature A. baumannii biofilm. Sytox Green reagent achieved 100% membrane permeation of Gram-positive and Gram-negative bacteria. The RK8 peptide did not show cytotoxic effects against murine macrophages (64 μM), erythrocytes (100 μM) or Galleria mellanella larvae (960 μM). In the stability test against peptidases, the RK8 peptide was stable, maintaining around 60% of the molecule intact after 120 min of incubation. These results highlight the potential of RK8 to be a promising strategy for developing a new antimicrobial and antibiofilm agent, inspiring and motivating further research in antimicrobial peptides.

Impedance Spectroscopy Unveiled the Surfactant-Induced Unfolding and Subsequent Refolding of Human Serum Albumin.

Protein-surfactant interaction is a dynamic interplay of electrostatic and hydrophobic forces that ensues from the folding of a protein. We employ impedance spectroscopy (IS), a label-free method, to investigate the unfolding and refolding of human serum albumin (HSA), a globular plasma protein, in the presence of two surfactants: polysorbate-20 (Tween-20), a nonionic surfactant, and sodium dodecyl sulfate (SDS), an anionic surfactant. The equivalent electrical analog circuit was predicted from impedance spectra of HSA in an aqueous solution at physiological pH and room temperature, focusing on varying the concentration of codissolved surfactants. A change in the dielectric constant (ε') and ionic conductivity (κ) is observed by comparing the surfactant-treated protein samples to the bare surfactant solutions to assess the conformational changes induced by surfactants in HSA. Far-UV circular dichroism analysis revealed a decrease in α-helices and an increase in β-sheets and random coils upon SDS addition, which were reversed by Tween-20. Dynamic light scattering supported the findings by measuring changes in the hydrodynamic diameter (dh) of HSA. Unfolding and refolding of HSA with surfactants were also observed through photoluminescence spectroscopy by examining the microenvironment surrounding the single tryptophan (W) within the protein, and the thermodynamic parameters were obtained using the modified Stern-Volmer equation. Our research explores the intriguing domain of protein-surfactant interactions, offering insights with promising applications across diverse biological processes and IS as a suitable alternative technique for investigating protein conformational changes by studying the electrical response of the samples.

Polarized Emission in Chiral Quasi‐2D Perovskite Light‐Emitting Diode

AbstractChiral 2D perovskites synthesized using (R‐+)‐α‐Methylbenzylamine (R‐MBA) and (S–)‐α‐Methylbenzylamine (S‐MBA) as barrier molecules exhibit chirality at low n values, as confirmed by circular dichroism (CD) analysis. The Anisotropy factor decreases significantly with increasing the n value, reaching 0.0032 for pure 2D perovskite. These synthesized quasi‐2D chiral perovskites have been successfully integrated into light‐emitting diodes (LEDs), demonstrating polarized emission for the first time. Remarkably, n = 3, R enantiomer achieves a total external quantum efficiency (EQE) of 12.1%, with polarized EQE of 4%. In the case of n = 2, the total EQE for both enantiomers is 4.3%, with polarized EQE of 3.4%, higher than the non‐polarized EQE of 0.9%. Degree of Polarization (DOP) is employed to characterize the fraction of an electromagnetic wave that exhibits polarization. The DOP analysis reveals a significant increase for low dimensional perovskite. Particularly for n = 2, R enantiomer exhibits a DOP of 53% compared to 2.5% for the non‐chiral molecule BzA. These findings demonstrate efficient polarized emission in 2D chiral perovskite LEDs, indicating promising avenues for advanced optical device applications.

Intracellular bactericidal activity and action mechanism of MDP1 antimicrobial peptide against VRSA and MRSA in human endothelial cells.

Staphylococcus aureus is a prominent cause of postoperative infections, often persisting within host cells, leading to chronic infections. Conventional antibiotics struggle to eliminate intracellular S. aureus due to poor cell penetration. Antimicrobial peptides are a new hope for tackling intracellular bacteria. Accordingly, this study examines the antimicrobial peptide MDP1, derived from melittin, for its efficacy against intracellular S. aureus. In this study, the physiochemical properties (Prediction of three-dimensional structure, circular dichroism and helical wheel projection analysis) were investigated. Extracellular antibacterial activity and cytotoxicity of MDP1 were also assessed. The mechanism of interaction of MDP1 with S. aureus was evaluated by molecular dynamic simulation, atomic force and confocal microscopy. Bacterial internalization into an endothelial cell model was confirmed through culture and transmission electron microscopy. The effect of the peptide on intracellular bacteria was investigated by culture and epi-fluorescence microscopy. 3D structural prediction proved the conformation of MDP1 as an α-helix peptide. Helical-wheel projection analysis indicated the proper orientation of hydrophobic amino acid residues for membrane interaction. CD spectroscopy of MDP1 showed that MDP1 in SDS 10 and 30 mM adopted 87 and 91% helical conformation. Atomic force and confocal microscopy assessments as well as molecular dynamics studies revealed the peptide-bacterial membrane interaction. MDP1, at the concentration of 0.32 μg mL-1, demonstrated a fold reduction of 21.7 ± 1.8, 1.7 ± 0.2, and 7.3 ± 0.8 in intracellular bacterial load for ATCC, VRSA, and MRSA, respectively. Molecular dynamics results demonstrate a preferential interaction of MDP1 with POPG/POPE membranes, primarily driven by electrostatic forces and hydrogen bonding. In POPC systems, two out of four MDP1 interacted effectively, while all four MDP1 engaged with POPG/POPE membranes. Gathering all data together, MDP1 is efficacious in the reduction of intracellular VRSA and MRSA proved by culture and epi-fluorescent microscopy although further studies should be performed to increase the intracellular activity of MDP1.

The interaction between resveratrol and catalase was studied by multispectral method and molecular docking simulation

In this research, we delved into the interaction mechanism between Resveratrol (RES) and Catalase (CAT) utilizing a suite of advanced analytical techniques, including multispectral analysis and molecular docking simulation. A comprehensive assessment of their binding characteristics, modalities, distances, and the subsequent impact on CAT's secondary structure was conducted through fluorescence spectroscopy, ultraviolet spectroscopy, circular dichroism, three-dimensional fluorescence spectroscopy, synchronous fluorescence spectroscopy, and molecular docking simulation. By employing the Stern-Volmer equation to determine KSV and Kq values, Our findings indicate that the RES-CAT interaction results in static fluorescence quenching, By conducting experiments, the binding constant KA and the number of binding sites n for CAT and RES were determined, with the number of binding sites n being close to 1, indicating the presence of a specific binding site between CAT and RES. Parameters at 298 K. The negative values of ΔH and ΔS suggest that van der Waals forces and hydrogen bonding are predominant in mediating the interaction between CAT and RES. The negativity of ΔG indicates the spontaneity of the reaction. The Fӧrster resonance energy transfer theory calculation revealed a binding distance of 4.917 nm, substantiating the occurrence of non-radiative energy transfer between RES and CAT. Furthermore, the UV-VIS absorption spectrum, synchronous fluorescence spectrum, three-dimensional fluorescence spectrum, and circular dichroism analyses collectively demonstrated that RES induces alterations in CAT's secondary structure, augmenting the polarity around tryptophan (Trp) residues, reducing hydrophobic interactions, and bolstering hydrophilicity. This work offers novel insights into the antioxidant mechanisms of resveratrol within biological systems and holds significant theoretical and practical implications for the medical application and development of resveratrol.