Covalent Conjugation Research Articles

Aspergillus flavipes L-methionine γ-lyase-β-cyclodextrin conjugates with improved stability, catalytic efficiency and anticancer activity

AbstractAspergillus flavipes L-methionine γ-lyase (MGL) has been authenticated as a powerful anticancer agent towards various solid tumors, however, the catalytic efficiency and stability of this enzyme remains the main challenge for its further in vivo applications. Thus, the objective of this study was to enhance the catalytic efficiency, structural stability of A. flavipes MGL, in addition to boost their anticancer activity, via conjugation with β-cyclodextrin. The purified A. flavipes MGL was (38.1 μmol/mg/min) was conjugated with β-cyclodextrin, with immobilization yield 80%. The conjugation process of MGL with β-cyclodextrin was verified from the FTIR analysis, molecular docking analysis, ensuring the covalent conjugation process via the hydrogen, and hydrophobic interactions with the cyclodextrin hydroxyl groups and MGL surface amino acids residues. The free and CD-MGL have the same optimum reaction temperature 37 °C, reaction pH 7.5 and pH stability pH 6.5–8.0. The CD-MGL conjugates had a significant stability to proteinase K and trypsin digestion. The affinity of CD-MGL was increased by ~ 2 folds to L-methionine (KM 3.1 mM), compared to the free one (KM 7.2 mM), as well as the catalytic efficiency of MGL was increased by 1.8 folds upon cyclodextrin conjugation. The higher affinity of CD-MGL for L-methionine might be due to re-orientation of the MGL to bind with the substrate by multiple interactions hydrogen, hydrophobic and covalent bonds compared to the free one. The thermal stability of MGL was increased by ~ 2 folds for the tested treatments, upon cyclodextrin conjugation. The in vitro anticancer activity of CD-MGL was enhanced by 2 folds against the HCT-116 (IC50 value 13.9 μmol/mg/min) and MCF7 (IC50 value 9.6 μmol/mg/min), compared to the free MGL (~ 21.4 μmol/mg/min). The enzymes displayed a significant activity against the proliferation of Ehrlich ascites carcinoma in vivo, with an obvious improvement on the liver tissues, as revealed from the histopathological sections

High-Throughput Discovery of Substrate Peptide Sequences for E3 Ubiquitin Ligases Using a cDNA Display Method.

Cells utilize ubiquitin as a posttranslational protein modifier to convey various signals such as proteasomal degradation. The disfunction of ubiquitylation or following proteasomal degradation can give rise to the accumulation and aggregation of improperly ubquitylated proteins, which is known to be a general causation of many neurodegenerative diseases. Thus, the characterization of substrate peptide sequences of E3 ligases is crucial in biological and pharmaceutical sciences. In this study, we developed a novel high-throughput screening system for substrate peptide sequences of E3 ligases using a cDNA display method, which enables covalent conjugation between peptide sequences and their corresponding cDNA sequences. First, we focused on the MDM2 E3 ligase and its known peptide substrate as a model to establish the screening method, and confirmed that cDNA display method was compatible with in vitro ubiquitylation. Then, we demonstrated identification of MDM2 substrate sequences from random libraries to identify a novel motif (VKFTGGQLA). Bioinformatics analysis of the hit sequences was performed to gain insight about endogenous substrate proteins.

Structure-Based Design of Covalent SARS-CoV-2 Papain-like Protease Inhibitors.

The COVID-19 pandemic is caused by SARS-CoV-2, a highly transmissible and pathogenic RNA betacoronavirus. Like other RNA viruses, SARS-CoV-2 continues to evolve with or without drug selection pressure, and many variants have emerged since the beginning of the pandemic. The papain-like protease, PLpro, is a cysteine protease that cleaves viral polyproteins as well as ubiquitin and ISG15 modifications from host proteins. Leveraging our recently discovered Val70Ub binding site in PLpro, we designed covalent PLpro inhibitors by connecting cysteine reactive warheads to the biarylphenyl PLpro inhibitors via flexible linkers. Several leads displayed potent enzymatic inhibition (IC50 = 0.1-0.3 μM) and antiviral activity (EC50 = 0.09-0.96 μM). Fumaramide inhibitors Jun13567 (15), Jun13728 (16), and Jun13714 (18) showed favorable in vivo pharmacokinetic properties with intraperitoneal injection. The X-ray crystal structure of PLpro with Jun13567 (15) validated our design strategy, revealing covalent conjugation between the catalytic Cys111 and the fumaramide warhead. The results suggest these covalent PLpro inhibitors are promising SARS-CoV-2 antiviral drug candidates.

A theranostic photosensitizer-conjugated albumin co-loading with resiquimod for cancer-targeted imaging and robust photo-immunotherapy

Cancer immunotherapy remains a low immune response rate in clinic because of dominant immunosuppressive tumor microenvironment (TME) and lack of effective drug to specifically remodel the TME. In this work, we introduced a tumor-seeking human serum albumin (HSA) based delivery platform by covalently conjugating with a tumor-targeting near-infrared (NIR) photosensitizer (IR-DBI) and non-covalently loading of immune modulator Resiquimod (R848). HSA exhibited tumor-preferential accumulation after covalent conjugation with IR-DBI. Meanwhile, HSA restricted the rotation of IR-DBI, narrowed the HOMO-LUMO energy gap, significantly enhanced fluorescent intensity and dual-modal phototherapy (PTT/PDT). The enhanced phototherapeutic effect further induced robust ICD effect. More importantly, non-covalent loading of R848 could be released from HSA at tumor sites by laser irradiation-induced heat. The in-situ release of R848 in TME efficiently promoted the maturation of DC cells and repolarized M2 macrophages to M1 macrophages. Consequently, robust photo-induced antitumor immunity was triggered in the different mice models bearing primary and distant tumors or lung metastasis, which was further enhanced by combining with CTLA-4 blockade therapy. Taken together, this work may present a versatile albumin composite which exhibits tumor-preferential accumulation and imaging-guided PDT/PTT/immunotherapy.

Surface-induced self-assembly of peptides turns superhydrophobic surface of electrospun fibrous into superhydrophilic one

Current surface modification strategies for electrospun materials always require covalent conjugation technology, which is relatively inefficient and might damage the bioactivity and structure of peptides and proteins. Here we introduce the use of surface-induced self-assembly technology to modify electrospun materials, which is a simple but efficient noncovalent-based process. Results show that the peptide NapFFGRGD forms burr-like structures on the surface of PCL fibers, reducing the water contact angle of the fibers. Adjusting the peptide sequence and salt concentration affects the self-assembly and surface properties of modified PCL fibers. Additionally, we demonstrate the potential application of this surface modification technique for enhancing cellular responses in tissue engineering applications. The research provides valuable insights into the surface modification of PCL fibers and offers a new method for improving the biological compatibility of materials in tissue engineering.

Enhancing Antibacterial Properties of Titanium Implants through Covalent Conjugation of Self-Assembling Fmoc-Phe-Phe Dipeptide on Titania Nanotubes.

Bacterial infections and biofilm formation are significant challenges for medical implants. While titanium nanotube engineering improves biocompatibility, it cannot prevent bacterial adhesion and biofilm formation. Optimizing the biomaterial's surface chemistry is vital for its desired functioning in the biological environment. This study demonstrates the covalent conjugating of the self-assembling dipeptide N-fluorenylmethyloxycarbonyl-diphenylalanine (Fmoc-FF) onto titanium nanotube surfaces (TiNTs) without altering the topography. Fmoc-FF peptides, in conjugation with TiNTs, can inhibit biofilm formation, eradicate pre-existing biofilms, and kill bacteria. This functionalization imparts antibacterial properties to the surface while retaining beneficial nanotube topography, synergistically enhancing bioactivity. Surface characterization by XPS, FT-IR, EDS, and SEM confirmed the successful functionalization. Bacterial adhesion experiments showed a significantly improved antibacterial activity of the functionalized TiNT surfaces. This study opens future possibilities for associating biomedical applications such as cell-cell interactions, tissue engineering, and controlled drug delivery of multifunctional self-assembling short peptides with implant materials through surface functionalization.

A Fentanyl Hapten‐Displaying Lipid Nanoparticle Vaccine that Non‐Covalently Encapsulates a TLR7/8 Agonist and T‐helper Epitope Induces Protective Anti‐Fentanyl Immunity

Opioid use disorder ‐ particularly involving fentanyl ‐ has precipitated a public health crisis characterized by a significant increase in addiction and overdose‐related deaths. Fentanyl‐specific immunotherapy, which aims at inducing fentanyl‐specific antibodies capable of binding fentanyl molecules in the bloodstream, preventing their entry in the central nervous system, is therefore gaining momentum. Conventional opioid designs rely on the covalent conjugation of fentanyl analogues to immunogenic carrier proteins that hold the inherent capacity of mounting immunodominant responses. Here, we present an alternative fentanyl vaccine design that utilizes a non‐covalent assembly of lipid nanoparticles (LNPs) to deliver fentanyl haptens in conjunction with a CD4+ T‐helper peptide epitope and an imidazoquinoline TLR7/8 agonist. Our results demonstrate that a single intramuscular administration of the LNP‐based nanovaccine elicits fentanyl‐specific antibodies, significantly mitigating the effects of opioid overdose in preclinical mouse models. Furthermore, we analyzed the immunobiological behavior of the vaccine in vivo in mouse models, providing evidence that covalent attachment of a fentanyl hapten to a carrier proteins or peptide epitope is not necessary for inducing an effective immune response. However, co‐delivery ‐ specifically, the physical assembly of all immune cues into an LNP ‐ remains essential for inducing hapten‐specific immunity.

A Fentanyl Hapten-Displaying Lipid Nanoparticle Vaccine that Non-Covalently Encapsulates a TLR7/8 Agonist and T-helper Epitope Induces Protective Anti-Fentanyl Immunity.

Opioid use disorder - particularly involving fentanyl - has precipitated a public health crisis characterized by a significant increase in addiction and overdose-related deaths. Fentanyl-specific immunotherapy, which aims at inducingfentanyl-specificantibodies capable of binding fentanyl molecules in the bloodstream, preventing their entry in the central nervous system, is therefore gaining momentum.Conventional opioid designsrely on the covalent conjugation of fentanyl analogues to immunogenic carrier proteins that hold the inherent capacity of mounting immunodominant responses. Here, we present an alternative fentanyl vaccine design that utilizes a non-covalent assembly of lipid nanoparticles (LNPs) to deliver fentanyl haptens in conjunction with a CD4+T-helper peptide epitope and an imidazoquinoline TLR7/8 agonist. Our results demonstrate that a single intramuscular administration of the LNP-based nanovaccine elicits fentanyl-specific antibodies, significantly mitigating the effects of opioid overdose in preclinical mouse models. Furthermore, we analyzed the immunobiological behavior of the vaccine in vivo in mouse models, providing evidence that covalent attachment of a fentanyl hapten to a carrier proteins or peptide epitope is not necessary for inducing an effective immune response. However, co-delivery - specifically, the physical assembly of all immune cues into an LNP - remains essential for inducing hapten-specific immunity.

Hydrogen Sulfide (H2S) Activatable Photodynamic Therapy Against Colon Cancer by Tunable FRET Effect.

Developing activatable photodynamic agents is becoming a promising mode for realizing selective photodynamic therapy (PDT) in cancer treatment. However, till now only a few H2S-activatable photodynamic agents have been involved in this field. Here, we fabricated H2S-activatable nano-assemblies (P@TPPCY) for the treatment of colon cancer containing high concentration H2S. The H2S-activatable photosensitizer (TPPCY) was synthesized by covalent conjugation between tetraphenylporphyrin (TPP) and heptamethine cyanine (Cy7), and then TPPCY was encapsulated by an amphiphilic polymer DSPE-mPEG to form P@TPPCY nano-assemblies. We demonstrated that the photosensitizing effect of TPP was efficiently quenched by Cy7 with strong absorption in the NIR region via fluorescence resonance energy transfer (FRET) effect. Interestingly, the FRET effect between Cy7 and TPP was attenuated by the decrease of absorption of Cy7 in the NIR region after the Cy7 reacted with H2S, and then the photosensitizing effect of TPP in P@TPPCY was activated. Strikingly, the P@TPPCY showed efficient H2S-activatable photodynamic therapy (PDT) in vitro against HCT116 cells (human colon carcinoma cell line), thus this work provides a new method for realizing accurate treatment of colon cancer in virtue of high H2S concentration microenvironment.

Inclusion Complexation of Remdesivir with Cyclodextrins: A Comprehensive Review on Combating Coronavirus Resistance—Current State and Future Perspectives

Cyclodextrin (CD) derivatives have gained significant attention in biomedical applications due to their remarkable biocompatibility, unique inclusion capabilities, and potential for functionalization. This review focuses on recent advancements in CD-based assemblies, specifically their role in improving drug delivery, emphasizing remdesivir (RMD). The review introduces CD materials and their versatile applications in self-assembly and supramolecular assembly. CD materials offer immense potential for designing drug delivery systems with enhanced activity. Their inherent inclusion capabilities enable the encapsulation of diverse therapeutic agents, including RMD, resulting in improved solubility, stability, and bioavailability. The recent advances in CD-based assemblies, focusing on their integration with RMD have been concentrated here. Various strategies for constructing these assemblies are discussed, including physical encapsulation, covalent conjugation, and surface functionalization techniques. Furthermore, exploring future directions in these fields has also been provided. Ongoing research efforts are directed toward developing novel CD derivatives with enhanced properties, such as increased encapsulation efficiency and improved release kinetics. Moreover, the integration of CD-based assemblies with advanced technologies such as nanomedicine and gene therapy holds tremendous promise for personalized medicine and precision therapeutics

Recent Advances in Self-Assembling Peptide-Based Nanomaterials for Enhanced Photodynamic Therapy.

Self-assembling peptide-based materials with ordered nanostructures possess advantages such as good biocompatibility and biodegradability, superior controllability, and ease of chemical modification. Through covalent conjugation or non-covalent encapsulation, photosensitizers (PSs) can be carried by self-assembling peptide-based nanomaterials for targeted delivery towards tumor tissues. This improves the stability, solubility, and tumor accumulation of PSs, as well as reduces their dark toxicity. More importantly, these nanomaterials can be tailored with responsiveness to tumor microenvironment, which enables smart release of PSs for precise and enhanced photodynamic therapy (PDT). In this review, the self-assembly of peptide from the perspective of driving forces is first described, and various self-assembling peptide materials with zero to 3D nanostructures are subsequently highlighted for PDT of cancers in recent years. Finally, an outlook in this field is provided to motivate fabrication of advanced PDT nanomaterials.

Structural modification and functional improvement of lactoferrin through non-covalent and covalent binding to coffee polyphenol

Studies have suggested that milk may enhance or neutralize the bioavailability of coffee polyphenols, possibly due to reversible and irreversible interactions between coffee polyphenols and milk proteins. The effects of non-covalent and covalent binding of lactoferrin (BLF) to caffeic acid (CAA) on protein structure and function were investigated. SDS-PAGE analysis confirmed the covalent interaction between BLF and CAA. Multispectral experiments characterized the BLF-CAA complexes and conjugates, revealing alterations in the tertiary structure of the proteins in the BLF-CAA conjugates. Molecular docking and kinetics results demonstrated that hydrogen bonding, electrostatic interaction, and hydrophobic forces were the primary internal forces between CAA and BLF. When combined with CAA, the covalent conjugates exhibited superior functional properties including solubility, oxidation resistance, thermal stability, emulsification and foamability, bioaccessibility, and antimicrobial properties. This study offers a theoretical foundation and technical benchmark for the preparation of protein-based delivery vectors with synergistic effects.

Short DNAzymes for Efficient Catalysis of "Click" Reactions in Solution and on Surface.

We have systematically investigated and found surprising superior catalytic activities of very short DNAzymes for copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC), both in solution and on surface. As a key reaction of the "click chemistry" class, CuAAC is a highly efficient and specific covalent conjugation tool with demonstrated applications in organic synthesis, bioconjugation, and surface functionalization; however, it requires the presence of the Cu(I) catalyst, which is an unstable species in aqueous solutions. We show here that one ultrashort, 14-nucleotide-truncated fragment of an earlier in vitro selected DNAzyme (CLICK-17) shows a striking and superior catalytic activity toward the in trans CuAAC reaction in solution and on surface in the presence of either Cu(I) or Cu(II), at significantly lowered concentrations. These results obviate the need for long-sequence DNAzymes, selected out of the homogeneous solution phase, for application in complex surface environments.

A surfactant-mediated dynamic protection strategy for 100% waterborne fabrication of durable superhydrophobic coatings

Waterborne preparation of superhydrophobic coatings now becomes strongly favored because of its environmental and health friendliness. However, the huge surface energy difference between low surface-energy components and water still poses a challenge for achieving their co-dispersion. Here, a surfactant-mediated protection strategy is proposed to achieve well dispersion of both SiO2 nanoparticles and perfluorosilane in water as well as block their conjugation, thereby fabricating a 100% waterborne F-FC-SiO2 suspension with high homogeneity and dispersion stability. During the painting and drying process, demulsification spontaneously triggers the deprotection and enables sufficient covalent conjugation between perfluorosilane and SiO2, thus achieving a superhydrophobic coating. This supramolecular protection/deprotection approach enables high compatibility of the F-FC-SiO2 suspension with versatile waterborne polymer emulsions (e.g., waterborne polyurethane, waterborne polysiloxane, and waterborne epoxy resin) to fabricate durable superhydrophobic coatings. The resulting composite coating can impart super-repellency to various substrates against aqueous solutions and multiple oils, displaying high antifouling and self-cleaning properties, as well as resistance to extreme mechanical abrasion and weathering. This work provides a promising approach for the convenient production of totally waterborne superhydrophobic coatings.

Synthesis and in vitro study of surface-modified and anti-EGFR DNA aptamer -conjugated chitosan nanoparticles as a potential targeted drug delivery system

Nowadays, finding effective approaches for cancer therapy is one of the significant issues related to human health all over the world. Hence, in this research, we designed and synthesized a novel targeted DDS based on surface-modified chitosan (CS) for the effective delivery of noscapine (NO). As the surface of CS nanoparticles was firstly modified with carboxyl groups and followed by covalent conjugation of DNA-aptamer (Ap) as targeting and receptor blocker agent. Secondly, NO, as a chemotherapeutic agent, was loaded into prepared nano-complex and synthetics were effectively characterized via various analytical devices, including FT-IR, 1H NMR, DLS, Zeta potential analyzer, TGA, TEM, and SEM to verify quality and quantity of synthetics. Drug loading was obtained about 25 % and sustained drug release was observed for nano-complex at different pHs. Then, the cell viability assay was performed on MCF-7 (as breast cancer cell) and HFF-1 (as normal cell) cell lines to investigate cancer cell inhibition potency of nano-complex. Cell viability of cancer cells was 19.84 ± 1.87 % (for C-CS-Ap-NO) and 75.43 ± 2.64 % (for C-CS-Ap) after 72 h of treatment with 400 nM concentration. These results have been confirming the excellent potency of synthesized novel nano-complex as practical DDS in cancer therapy.

Design and Preparation of Lifetime-Based Dual Fluorescent/Phosphorescent Sensor of pH and Oxygen and its Exploration in Model Physiological Solutions and Cells.

In the present report, a novel dual pH-O2 sensor based on covalent conjugate of rhodamine 6G and cyclometalated iridium complex with poly(vinylpyrrolidone-block-vinyltetrazole) copolymer is reported. In model physiological solutions the sensor chromophores display independent phosphorescent and fluorescent lifetime responses onto variations in oxygen concentration and pH, respectively. Colocalization studies on Chinese hamster ovary cells demonstrate the preferential localization in endosomes and lysosomes. The fluorescent lifetime imaging microscopy-phosphorescent lifetime imaging microscopy (FLIM-PLIM) experiments show that the phosphorescent O2 sensor provides unambiguous information onto hypoxia versus normoxia cell status as well as semi-quantitative data on the oxygen concentration in cells in between these two states. However, the results of FLIM measurements indicate that dynamic lifetime interval of the sensor (≈0.5ns between pH values 5.0 and 8.0) is insufficient even for qualitative estimation of pH in living cells because half-width of lifetime distribution in the studied samples is higher than the sensor dynamic interval. Nevertheless, the variations in rhodamine emission intensity are much higher and allow rough discrimination of acidic and neutral cell conditions. Thus, the results of this study indicate that the suggested approach to the design of dual pH-O2 sensors makes possible to prepare the biocompatible and water-soluble conjugate with fast cellular uptake.

SYNTHESIS OF CHITOSAN COMPOSITES WITH METHYLENE BLUE, MALACHITE GREEN AND ACID FUCHSIN DYES FOR ENHANCEMENT OF THEIR ANTIMICROBIAL ACTION

The effect of chitosan oxidation with sodium periodate on the ability to form complexes with the dyes methylene blue, malachite green and acid fuchsin, which have an antiseptic effect, was investigated. A non-covalent interaction occurred for methylene blue and malachite green, while a covalent interaction occurred for acid fuchsin. Fourier-transform infrared spectroscopy confirmed the formation of chitosan composites with these dyes. The antimicrobial effect of these dyes complexed with oxidised chitosan against Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Escherichia coli was investigated. The non-covalent attachment of malachite green and methylene blue to chitosan preserved their antimicrobial effect; hence, complexes of these dyes with oxidised chitosan provide a more convenient form that is less staining. However, the covalent conjugation of acid fuchsin with oxidised chitosan caused a significant loss of the antimicrobial activity. The developed method of conjugation of specific dyes with chitosan can be used to create complexes of this biopolymer with a wide range of biologically active organic substances.

Covalent Grafting of Tanfloc on Titania Nanotube Arrays: An Approach to Mitigate Bacterial Adhesion and Improve the Antibacterial Efficacy of Titanium Implants

AbstractImplanted medical devices often face the challenge of infections, which can compromise their successful integration and use. To address this issue, this study demonstrates the covalent grafting of a tannin‐based antimicrobial biopolymer tanfloc (TAN) onto the titania nanotube arrays (TiNTs) surface to enhance antibacterial properties. Due to its polyphenolic and ionic structural configuration, tanfloc possesses unique properties that enable it to interact with and disrupt bacterial cell walls and membranes. Combining the topographical effect of TiNTs with the inherent antibacterial properties of tanfloc, this approach aims to mitigate bacterial threats on medical implants effectively. The successful attachment of tanfloc on TiNTs is confirmed through X‐ray photoelectron spectroscopy (XPS) and Fourier‐transform infrared spectroscopy (FT‐IR). The antibacterial and antibiofilm efficacy of the tanfloc‐functionalized TiNTs is evaluated against Staphylococcus aureus (Gram‐positive) and Pseudomonas aeruginosa (Gram‐negative) bacteria. The findings suggest that the covalent conjugation of tanfloc onto TiNTs is a promising approach to improve the infection resistance of titanium‐based medical implants, with potential applications in orthopedic, dental, and other biomedical device areas.

Protein SUMOylation and Its Functional Role in Nuclear Receptor Control

Post-translational protein modifications (PTMs) significantly enhance the functional diversity of proteins and are therefore important for the expansion and the dynamics of the cell’s proteome. In addition to structurally simpler PTMs, substrates also undergo modification through the reversible attachment of small proteins. The best understood PTM of this nature to date is the covalent conjugation of ubiquitin and ubiquitin-like proteins (UBLs) to their substrates. The protein family of small ubiquitin-like modifier (SUMO) is one of these UBLs that has received increasing scientific attention. The pathway of SUMOylation is highly conserved in all eukaryotic cells and is crucial for their survival. It plays an essential role in many biological processes, such as the maintenance of genomic integrity, transcriptional regulation, gene expression, and the regulation of intracellular signal transduction, and thereby influences DNA damage repair, immune responses, cell cycle progression, and apoptosis. Several studies have already shown that in this context protein SUMOylation is involved in the control mechanisms of various cellular receptors. This article unites data from different studies focusing on the investigation of the strictly conserved three-step enzyme cascade of protein SUMOylation and the functional analysis of the involved proteins E1, E2, and E3 and SUMOylation target proteins. Furthermore, this review highlights the role of nuclear receptor SUMOylation and its importance for the cellular functionality and disease development arising from defects in correct protein SUMOylation.

Enhancing the water dispersibility and intestinal targeting of pterostilbene using tannic acid-whey protein conjugates

In this study, three types of whey protein isolate (WPI)-tannic acid (TA) covalent conjugates (WPI-TA) were synthesized via alkaline treatment, free radical-induced method, and laccase induction. These conjugates were subsequently utilized in the antisolvent precipitation method to encapsulate pterostilbene (Pte). The structural and functional properties of these conjugates as carriers for intestinal-targeted Pte delivery were thoroughly evaluated. SDS-PAGE, total phenolic content measurement, grafting efficiency, and FT-IR analysis confirmed the successful formation of three distinct covalent conjugates. Among them, the laccase-induced WPI-TA conjugates exhibited the highest TA grafting efficiency. The particle size of this conjugate was measured at 115.25 nm with a PDI of 0.31. Upon encapsulation of Pte, this conjugate exhibited superior thermal stability, enhanced antioxidant properties, and sustained release within the gastrointestinal tract. The water solubility of Pte in these nanoparticles was found to be 361.19 times higher than that of free Pte, presenting a promising strategy for targeted intestinal delivery of Pte.