Dearomatization drives complexity generation in freshwater organic matter.

Dissolved organic matter (DOM) is one of the most complex, dynamic and abundant sources of organic carbon, but its chemical reactivity remains uncertain1-3. Greater insights into DOM structural features could facilitate understanding its synthesis, turnover and processing in the global carbon cycle4,5. Here we use complementary multiplicity-edited 13C nuclear magnetic resonance (NMR) spectra to quantify key substructures assembling the carbon skeletons of DOM from four main Amazon rivers and two mid-size Swedish boreal lakes. We find that one type of reaction mechanism, oxidative dearomatization (ODA), widely used in organic synthetic chemistry to create natural product scaffolds6-10, is probably a key driver for generating structural diversity during processing of DOM that are rich in suitable polyphenolic precursor molecules. Our data suggest a high abundance of tetrahedral quaternary carbons bound to one oxygen and three carbon atoms (OCqC3 units). These units are rare in common biomolecules but could be readily produced by ODA of lignin-derived and tannin-derived polyphenols. Tautomerization of (poly)phenols by ODA creates non-planar cyclohexadienones, which are subject to immediate and parallel cycloadditions. This combination leads to a proliferation of structural diversity of DOM compounds from early stages of DOM processing, with an increase in oxygenated aliphatic structures. Overall, we propose that ODA is a key reaction mechanism for complexity acceleration in the processing of DOM molecules, creation of new oxygenated aliphatic molecules and that it could be prevalent in nature.

pH-regulated Tannic acid and soybean protein isolate adhesive for enhanced performance in plant-based meat analogues.

A food adhesive comprising tannic acid (TA) and soybean protein isolate (SPI) was developed to establish a cohesive bond between soy protein gel and simulated fat. The impact of varying TA concentrations and pH levels on the adhesive's rheology, thermal stability, chemical structure, and tensile strength were investigated. Rheological results revealed a gradual decrease in adhesive viscosity with increasing TA content. Differential scanning calorimetry (DSC) and thermal gravimetric (TG) results indicated that the stability of the adhesive improved with higher TA concentrations, reaching its peak at 0.50% TA addition. The incorporation of TA resulted in the cross-linking of amino group in unfolded SPI molecules, forming a mesh structure. However, under alkaline conditions (pH 9), adhesive viscosity and stability increased compared to the original pH. This shift was due to the disruption of the SPI colloidal charge structure, an increase in the stretching of functional groups, further unfolding of the structure, and an enhanced binding of SPI to TA. Under the initial pH conditions, SPI reacted with TA's active site to form covalent crosslinked networks and hydrogen bonds. In alkaline condition, beyond hydrogen and ionic bonding, the catechol structure was oxidized, forming an ortho-quinone that crosslinked SPI and created a denser structure. Tensile strength measurements and freeze-thaw experiments revealed that the adhesive exhibited maximum tensile strength and optimal adhesion with 0.75% TA at pH 9, providing the best overall performance. This study provides a new formulation and approach for developing plant-based meat analogues adhesives.

Identification and characterization of silver nanoparticles from Erythrina indica and its antioxidant and Uropathogenic antimicrobial properties.

The plant Erythrina indica comes under Fabaceae family, mainly used for used in traditional medicine as nervine sedative, antiepileptic, antiasthmatic, collyrium in opthalmia, antiseptic. Current study focused synthesize of silver nanoparticles (AgNPs) by E. indica leaf ethanol extract. The green-synthesized AgNPs underwent characterization using multiple analytical techniques, including UV-visible, FTIR, DLS, SEM, TEM, XRD, and EDX, and estimation of their antioxidant activity and antimicrobial activity. Phytochemical analysis identified alkaloids, tannins, saponins, flavonoids, and phenols as secondary metabolites. The Total Phenol Content (TPC) was determined to be 237.35 ± 2.02 mg GAE-1, indicating a substantial presence of phenolic compounds. The presence of AgNPs was verified through UV-Visible analysis at 420 nm, and FT-IR revealed characteristic phenolic functional groups. DLS analysis indicated a narrow size distribution (polydispersity index - PDI: 3.47%), with SEM revealing spherical AgNPs of approximately 20 nm. TEM showed homogeneous, highly polycrystalline AgNPs with lattice spacing at 0.297. XRD analysis demonstrated crystallinity and purity, with distinct reflection peaks corresponding to miller indices of JCPDS card no. 01 087 1473. In vitro, AgNPs exhibited robust antioxidant activity like; DPPH, ABTS, and H2O2, surpassing E. indica-assisted synthesis. ABTS assay indicated higher antioxidant activity (81.94 ± 0.05%) for AgNPs at 734 nm, while E. indica extraction showed 39.67 ± 0.07%. At 532 nm, both E. indica extraction (57.71 ± 0.11%) and AgNPs (37.41 ± 0.17%) exhibited H2O2 scavenging. Furthermore, AgNPs displayed significant antimicrobial properties, inhibiting Staphylococcus aureus (15.7 ± 0.12 mm) and Candida albicans (10.7 ± 0.17 mm) byfor the concentration of 80 µg/mL. Through the characterizations underscore of the potential of Erythrina indica-synthesized AgNPs, rich in polyphenolic compounds, for pharmacological, medical, biological applications and antipyretic properties.

Development of a tannic acid- and silicate ion-functionalized PVA-starch composite hydrogel for in situ skeletal muscle repairing.

The repair capacity of skeletal muscle is severely diminished in massive skeletal muscle injuries accompanied by inflammation, resulting in muscle function loss and scar tissue formation. In the current work, we developed a tannic acid (TA)- and silicate ion-functionalized tissue adhesive poly(vinyl alcohol) (PVA)-starch composite hydrogel, referred to as PSTS (PVA-starch-TA-SiO32-). It was formed based on the hydrogen bonding of TA to organic polymers, as well as silicate-TA ligand interaction. PSTS could be gelatinized in minutes at room temperature with crosslinked network formation, making it applicable for injection. Further investigations revealed that PSTS had skeletal muscle-comparable conductivity and modulus to act as a temporary platform for muscle repairing. Moreover, PSTS could release TA and silicate ions in situ to inhibit bacterial growth, induce vascularization, and reduce oxidation, paving the way to the possibility of creating a favorable microenvironment for skeletal muscle regeneration and tissue fibrosis control. The in vivo model confirmed that PSTS could enhance muscle fiber regeneration and myotube formation, as well as reduce infection and inflammation risk. These findings thereby implied the great potential of PSTS in the treatment of formidable skeletal muscle injuries.

[Dynamic Chemistry of Tannins].

Tannins are a group of polyphenols that possess the ability to precipitate proteins, causing an undesirable astringent taste by interacting with salivary peptides. This interaction deactivates the digestive enzymes; therefore, tannins are considered as plant defense substances. The health benefits of tannins and related polyphenols in foods and beverages have been demonstrated by biological and epidemiological studies; however, their metabolism in living plants and the chemical changes observed during processing of foods and medicinal herbs raises some questions. This review summarizes our studies concerning dynamic changes observed in tannins. Ellagitannins present in the young leaves of Camellia japonica and Quercus glauca undergo oxidative degradation as the leaves mature. Similar oxidative degradation is also observed in whiskey when it is kept for aging in oak barrels, and in decaying wood caused by fungi in natural forests. In contrast, ellagitannins have been observed to undergo reduction in the leaves of Carpinus, Castanopsis, and Triadica species as the leaves mature. This phenomenon of reductive metabolism in leaves enabled us to propose a new biosynthetic pathway for the most fundamental ellagitannin acyl groups, which was also supported by biomimetic synthetic studies. Polyphenols undergo dynamic changes during the process of food processing. Catechin in tea leaves undergo oxidation upon mechanical crushing to generate black tea polyphenols. Though detailed production mechanisms of catechin dimers have been elucidated, structures of thearubigins (TRs), which are complex mixtures of oligomers, remain ambiguous. Our recent studies suggested that catechin B-ring quinones couple with catechin A-rings during the process of oligomerization.

Wine tannins and their aggregation/release with lipids and proteins: Review and perspectives for neurodegenerative diseases.

Tannins are amphiphilic molecules, often polymeric, which can be generally described as a core containing hydrophobic aromatic rings surrounded by hydroxyl groups. They have been known for millennia and are part of human culture. They are ubiquitous in nature and are best known in the context of wine and tea tasting and food cultures. However, they are also very useful for human health, as they are powerful antioxidants capable of combating the constant aggressions of everyday life. However, their mode of action is only just beginning to be understood. This review, using physicochemical concepts, attempts to summarize current knowledge and present an integrated view of the complex relationship between tannins, proteins and lipids, in the context of wine drinking while eating. There are many thermodynamic equilibria governing the interactions between tannins, saliva proteins, lipid droplets in food, membranes and the taste receptors embedded in them. Taste sensations can be explained using these multiple equilibria: for example, astringency (dry mouth) can be explained by the strong binding of tannin micelles to the proline-rich proteins of saliva, suppressing their lubricating action on the palate. In the presence of lipid droplets in food, the equilibrium is shifted towards tannin-lipid complexes, a situation that reduces the astringency perceived when consuming a tannic wine with fatty foods, the so-called "camembert effect". Tannins bind preferentially to taste receptors located in mouth membranes, but can also fluidify lipids in the non-keratinized mucous membranes of the mouth, which can impair the functioning of taste receptors there. Cholesterol, present in large quantities in keratinized mucous membranes, stiffens them and thus prevents tannins from disrupting the conduction of information through other taste receptors. As tannins assemble and disassemble depending on whether they are in contact with proteins, lipids or taste receptors, a perspective on their potential use in the context of neurodegenerative diseases where fibrillation is a key phenomenon will also be discussed.

Plant polyphenol surfactant construction with strong surface activity and chelation properties as efficient decontamination of UO22+ on cotton fabric.

Chemically synthesized surfactants have promising applications in the treatment of uranium, however, their hazardous environmental effects, non-biodegradability, and numerous drawbacks prevent them from being widely used in practice. Herein, we successfully synthesized a green chelating and foaming integrated surfactant (BTBS) by Mannich reaction and acylation of bayberry tannin for the effective removal of UO22+ from aqueous environments or solid surfaces. The as-prepared surfactant was systematically characterized by FT-IR, showing that the hydrophobic groups were successfully grafted onto tannin. The modified material showed better foaming and emulsifying properties, which proved this method could improve the amphiphilicity of tannin. Moreover, for the first time, a foam fractionation method in conjunction with a tannin-based surfactant was applied for UO22+ removal from water. This surfactant was used as a co-surfactant and could readily remove 90 % of UO22+ (20 mg L-1) from water. The removal of UO22+ could be completed in a short time (30 min), and the maximum adsorption capacity was determined as 175.9 mg g-1. This surfactant can also be used for efficient decontamination of uranium-contaminated cotton cloth with a high removal rate of 94.55 %. In addition, the mechanism studies show that the adsorption of BTBS for UO22+ can be mainly attributed to a chelating mechanism between UO22+ and the adjacent phenolic hydroxyls. The novel biomass-derived BTBS with advantages such as high capture capacity, environmental friendliness, and cost-effectiveness suggests that it plays an important role in the remediation of radionuclide pollution.

A multiple cross-linking strategy to develop an environment-friendly and water resistance wheat gluten protein wood adhesive.

Wheat gluten (WG) shows great promise to synthesize environment-friendly wood adhesives. However, their weak bonding strength and poor water resistance have limited its application in the commercial wood-based panel industry. In this study, a novel WG-based adhesive was developed by constructing a multiple cross-linking network generated by covalent and non-covalent bonds. The potential mechanism was revealed by FT-IR analysis. Furthermore, their surface morphology, thermal stability, viscosity, and residual rate of adhesives with different compositions were systematically characterized and compared. The results showed that the hydrogen bonding, reactions between amine groups and tannin, and ring opening reaction of epoxy, synergistically contributed to generate a highly crosslinked network. The wet/boil water strength of the plywood prepared from WG/tannin/ethylene imine polymer (PEI)-glycerol triglycidyl ether (GTE) adhesive with the addition of 15 % GTE could reach 1.21 MPa and 1.20 MPa, respectively, and a mildew resistance ability was observed. This study provides a facile strategy to fabricate high-performance plant protein-based adhesives with desirable water resistance for practical application.

Cold plasma treatment of tannic acid as a green technology for the fabrication of advanced cross-linkers for bioactive collagen/gelatin hydrogels.

Tannic acid (TA) is a natural compound studied as the cross-linker for biopolymers due to its ability to form hydrogen bonds. There are different methods to improve its reactivity and effectiveness to be used as a modifier for biopolymeric materials. This work employed plasma to modify tannic acid TA, which was then used as a cross-linker for fabricating collagen/gelatin scaffolds. Plasma treatment did not cause any significant changes in the structure of TA, and the resulting oxidized TA showed a higher antioxidant activity than that without treatment. Adding TA to collagen/gelatin scaffolds improved their mechanical properties and stability. Moreover, the obtained plasma-treated TA-containing scaffolds showed antibacterial properties and were non-hemolytic, with improved cytocompatibility towards human dermal fibroblasts. These results suggest the suitability of plasma treatment as a green technology for the modification of TA towards the development of advanced TA-crosslinked hydrogels for various biomedical applications.

Effects of Covalent or Noncovalent Binding of Different Polyphenols to Acid-Soluble Collagen on Protein Structure, Functionality, and Digestibility.

In this study, the structure, function, and digestibility of noncovalent complexes and covalent conjugates formed by acid-soluble collagen with polyphenols of different structures (quercetin, epicatechin, gallic acid, chlorogenic acid, procyanidin, and tannic acid) were investigated. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that polyphenols were covalently bound to collagen by laccase catalytic oxidation. Biolayer interferometry revealed that the noncovalent binding strength of polyphenols to collagen from high to low was quercetin > gallic acid > chlorogenic acid > epicatechin, which was consistent with the trend of covalent polyphenol binding. Procyanidin and tannic acid had strong noncovalent binding, but their covalent binding ability was weak. Compared with the pure collagen, the complexes improved emulsification and antioxidant properties (more than 2.5 times), and the conjugates exhibited better thermal stability (99.4-106.8 °C) and antidigestion ability (reduced by more than 37%). The finding sheds new light on the use of collagen as a functional food ingredient in the food industry.

Tannin complexation with metal ions and its implication on human health, environment and industry: An overview.

Tannins, also known as plant polyphenols (PPs), are secondary metabolites widely existing in higher plants and are a kind of natural renewable resource with wide distribution, variety and quantity. Tannin has become an important class of fine chemicals due to the easily modified molecular structure and the properties of antibacterial and antioxidant, combining with protein and complexing with metal ion. Besides being used for tanning leather, tannins are also widely used in wood adhesive, concrete water-reducing agents, oil drilling fluid viscosity-reducing agents, pharmaceutical, mineral processing, water treatment, gas desulfurization, metal anticorrosion, wood anticorrosion, printing and dyeing, liquor clarification, oil antioxidant, daily chemical products and other products preparation. There are two groups of tannins: condensed tannins (CTs) (flavonoid-derived proanthocyanidins) and hydrolysable tannins (HTs) (gallic acid ester-derived). Tannins can form complexes with metals through the ortho-dihydroxyphenolic group(s), especially with transition metals. The structure-activity relationships, stoichiometry, and origin of the insolubility of which were emphasized. Furthermore, this paper proposed an in-depth discussion of the associations of tannins-metal complexes in human health, environment and industries.

Why is it important to understand the nature and chemistry of tannins to exploit their potential as nutraceuticals?

Tannins comprise a large group of polyphenols that can differ widely in chemical composition and molecular weight. The use of tannins dates back to antiquity, but it is only in recent years that their potential use as nutraceuticals associated with the human diet is beginning to be exploited. Although the biological effects of these phytocomplexes have been studied for many years, there are still several open questions regarding their chemistry and biotransformation. The vastness of the molecules that make up the class of tannins has made their characterisation, as well as their nomenclature and classification, a daunting task. This review has been written with the aim of bringing order to the chemistry of tannins by including aspects that are sometimes still overlooked or should be updated with new research in order to understand the potential of these phytocomplexes as active ingredients or technological components for nutraceutical products. Future trends in tannin research should address many questions that are still open, such as determining the exact biosynthetic pathways of all classes of tannins, the actual biological effects determined by the interaction of tannins with other molecules, their metabolization, and the best extraction methods, but with a view to market requirements.

Antimicrobial films fabricated with myricetin nanoparticles and chitosan derivation microgels for killing pathogenic bacteria in drinking water.

Pathogenic bacteria in drinking water threaten human health and life. In the work, antimicrobial films composed of myricetin@tannic acid (My@TA) nanoparticles (NPs) and chitosan derivation microgels were developed to kill pathogenic bacteria in drinking water. Hydrophobic My was first made into water soluble My@TA NPs using a solvent exchange method with TA as stabilizer. Polymeric microgels of carboxymethyl chitosan (CMCS)/hydroxypropyltrimethyl ammonium chloride chitosan (HACC) were then fabricated with a blending method. CMCS&HACC/My@TA multilayer films were further deposited on the internal surface of PET bottles by using a layer-by-layer (LbL) assembly technique. The PET bottles coated with the films could effectively kill pathogenic bacteria in water such as S. aureus, E. coli, Staphylococcus epidermidis, Pseudomonas fluorescens, Listeria monocytogenes and methicillin resistant Staphylococcus aureus (MRSA). In addition, CMCS&HACC/My@TA films displayed good antioxidant activity, water resistance, and in vivo biocompatibility with heart, liver, spleen, lung and kidney organs. We believe that the container coated with CMCS&HACC/My@TA films can be applied to prevent microbial contamination of drinking water.

Alcohol-Induced Conformation Changes and Thermodynamic Signatures in the Binding of Polyphenols to Proline-Rich Salivary Proteins.

The first contact of polyphenols (tannins) with the human body occurs in the mouth, where they are known to interact with proline-rich proteins (PRPs). These interactions are important at a sensory level, especially for the development of astringency, but affect also various other biochemical processes. Employing thermodynamic measurements, fluorescence and CD spectroscopy, we investigated the binding process of the prototypical polyphenol ellagic acid (EA) to different IB-PRPs and BSA, also in the presence of ethanol, which is known to influence tannin-protein interactions. Binding of EA to BSA and the small peptide IB7-14 is weak, but very strong to IB9-37. The differences in binding strength and stoichiometry are due to differences in the binding motifs, which also lead to differences in the thermodynamic signatures of the binding process. EA binding to BSA is enthalpy-driven, whereas binding to both IB7-14 and IB9-37 is entropy-driven. The presence of 10 vol.% EtOH, as present in wines, increases the binding constant of EA with BSA and IB7-14 drastically, but not that with IB9-37; however, it changes the binding stoichiometry. These differences can be attributed to the effect of EtOH on the conformation dynamics of the proteins and to changes in hydration properties in alcoholic solution.

Sucrose-tannin-nanosilica hybrid bio-adhesive based on dual dynamic Schiff base and disulfide bonds with enhanced toughness and cohesion.

Herein, a high-performance sucrose-tannin bio-based adhesive is developed based on consisting of oxidized sucrose (OS), tannin acid (TA), SiO2 nanoparticles and 2,2'-disulfanediylbis (ethan-1-amine) (DBA) by a facile chemical cross-linking strategy. The OS-TA and OS-TA@SiO2 bio-based adhesives are characterized by XPS, FTIR, and 13C NMR, while the bonding performance is also investigated using shear strength test. Results show that the optimal formulation of OS-TA bio-based adhesive is a 2:1:1 mass ratio for OS: TA: DBA. When the mass fraction of SiO2 is 15 % and the solid content of main components is 50 %, the OS-TA@SiO2 bio-based adhesive has excellent bonding strength. Relative to OS-TA, the wet bonding strength of the OS-TA@SiO2 enhanced from 1.16 MPa to 1.85 MPa, while the dry bonding strength improved from 1.90 MPa to 2.50 MPa. The wood failure rate of the plywood fabricated by using the OS-TA@SiO2 bio-based adhesive reaches 80 %. Therefore, relying on the high flexibility of dynamic disulfide bonds, adding SiO2 nanoparticles into the adhesive system can facilitate greatly the mechanical interlocking effect and make the chemical cross-linking network more compact through the synergistic chemical interactions. This work provides new insights into producing green and renewable bio-based wood adhesives using sucrose and tannin.

Nanocellulose-based tannin-immobilized biosorbent for efficient copper ion removal.

Tannins, which are water-soluble polyhydroxyphenols found in plant structures, offer a sustainable alternative to inorganic products for treating effluents. To overcome their solubility in water, new methods have been developed to immobilize tannin on materials such as nanocellulose, resulting in tannin-based adsorbents. This study investigates the production of different types of tannin-based adsorbents immobilized on nanocellulose nanofibrils using glutaraldehyde as a crosslinking agent. The adsorbents were characterized for their morphology, chemical composition, tannin leaching, thermal stability, and copper adsorption capacity. The main results showed that tannin leaching from the adsorbents in water was greatly reduced with glutaraldehyde. Scanning electron microscopy images showed tannin coating on the surface of nanocellulose nanofibrils due to immobilization. The lyophilized adsorbent was effective in adsorbing Cu (II) within 1 min, with the adsorbed amount increasing gradually until stabilization after 45 min. Overall, this study suggests that tannin immobilized on nanofibrillated cellulose has the potential to offer a sustainable and renewable alternative for effluent treatment.

Development of high internal phase emulsions with noncovalent crosslink of soy protein isolate and tannic acid: Mechanism and application for 3D printing.

In this study, we propose a design strategy using soy protein isolate (SPI)-tannic acid (TA) complexes crosslinked through noncovalent interactions to develop high internal phase emulsions (HIPEs) for 3D printing materials. The results of Fourier transform infrared spectroscopy, intrinsic fluorescence, and molecular docking analyses indicated that the dominant interactions occurring between the SPI and TA were mediated by hydrogen bonds and hydrophobic interactions. The secondary structure, particle size, ζ-potential, hydrophobicity and wettability of SPI was significantly altered by the addition of TA. The microstructure of HIPEs stabilized by SPI-TA complexes exhibited more regular and even polygonal shapes, thereby allowing the protein to form a dense self-supporting network structure. When the concentration of TA exceeded 50 µmol/g protein, the formed HIPEs remained stable after 45 days of storage. Rheological tests revealed that the HIPEs exhibited a typical gel-like (G' > G'') and shear-thinning behavior, which contributed to preferable 3D printing behavior.

Tannins Can Have Direct Interactions with Anthelmintics: Investigations by Isothermal Titration Calorimetry.

Plant tannins are known for their anthelmintic and antiparasitic activities and have been increasingly studied to battle the ever-growing problem of anthelmintic resistance. While tannins have been shown to exhibit these activities on their own, one approach would be to use them as complementary nutrients alongside commercial anthelmintics. So far, research on the interactions between tannins and anthelmintics is limited, and few studies have reported both synergistic and antagonistic effects depending on the type of tannin and the method used. These interactions could either strengthen or weaken the efficacy of commercial anthelmintics, especially if tannin-rich diets are combined with anthelmintics used as oral drenches. To study these interactions, a series of hydrolysable tannins (HTs) was selected, and their direct interactions with thiabendazole (TBZ) were evaluated by isothermal titration calorimetry (ITC), which allowed the detection of the exothermic interaction but also the roles and significances of different structural features of HTs in these interactions. Our results show that HTs can have a direct interaction with the benzimidazole anthelmintic TBZ and that the interaction is strengthened by increasing the number of free galloyl groups and the overall molecular flexibility of HTs.

Development of protein-polyphenol particles to stabilize high internal phase Pickering emulsions by polyphenols' structure.

Protein-polyphenol colloidal particles are promising stabilizers for high internal phase Pickering emulsions (HIPPEs). However, the relationship between the structure of the polyphenols and its ability to stabilize HIPPEs has not been studied thus far. In this study, bovine serum albumin (BSA)-polyphenols (B-P) complexes were prepared, and their ability to stabilize HIPPEs was investigated. The polyphenols were bound to BSA via non-covalent interactions. Optically isomeric polyphenols formed similar bonds with BSA, whereas a greater number of trihydroxybenzoyl groups or hydroxyl groups in the dihydroxyphenyl moieties of polyphenols increased the B-P interactions. Polyphenols also reduced the interfacial tension and enhanced the wettability at the oil-water interface. The HIPPE stabilized by BSA-tannic acid complex exhibited the highest stability among the B-P complexes and resisted demixing and aggregation during centrifugation. This study promotes the potential applications of polyphenol-protein colloidal particles-stabilized HIPPEs in the food industry.

Tannic acid-based sustained-release system for protein drugs.

In recent years, protein drug development has gained momentum, and simple and facile controlled-release systems without loss of activity are required. Herein, we developed a sustained-release system for protein drugs by exploiting the "astringency" mechanism, namely insoluble precipitate formation by interacting with tannic acid. Tannic acid formed insoluble precipitates with various protein drugs, such as nisin, insulin, lysozyme, ovalbumin, hyaluronidase, and human immunoglobulin G, through hydrophobic interactions and hydrogen bonds. The lysozyme/tannic acid complex retained in vitro lytic activity. Precipitates of the insulin/tannic acid complex prolonged hypoglycemic effects without loss of activity after subcutaneous administration. The ovalbumin/tannic acid complex enhanced anti-ovalbumin antibody production induced by ovalbumin, which may be attributed to its sustained-release profile. Accordingly, tannic acid is useful as a simple and user-friendly drug delivery system for protein drugs.