Vertasile ferritin nanocages: Applications in detection and bioimaging.

Food safety and human health remain significant concerns in the food industry. Detecting food contaminants and diagnosing diseases are critical aspects. Ferritin, an iron storage protein widely found in nature, offers unique advantages. Its hollow protein nanocage structure, distinct interfaces, hydrophobic or hydrophilic channels, and B-C loop regions recognized by transferrin receptor 1 make ferritin versatile for detecting heavy metals, free radicals, and bioimaging both in vitro and in vivo. This review summarizes ferritin's general characteristics, its specific properties as biosensors, and its applications in food safety and in vivo imaging. It emphasizes not only ferritin's role in detecting heavy metals like mercury and chemical hazards but also its potential in early diagnosing chronic diseases such as tumors, macrophages, and kidney diseases. Further research into ferritin promises advancements in enhancing food safety and improving human health diagnostics.

Understanding the Role of Filamentous Actin in Food Quality: From Structure to Application.

Actin, a multifunctional protein highly expressed in eukaryotes, is widely distributed throughout cells and serves as a crucial component of the cytoskeleton. Its presence is integral to maintaining cell morphology and participating in various biological processes. As an irreplaceable component of myofibrillar proteins, actin, including G-actin and F-actin, is highly related to food quality. Up to now, purification of actin at a moderate level remains to be overcome. In this paper, we have reviewed the structures and functions of actin, the methods to obtain actin, and the relationships between actin and food texture, color, and flavor. Moreover, actin finds applications in diverse fields such as food safety, bioengineering, and nanomaterials. Developing an actin preparation method at the industrial level will help promote its further applications in food science, nutrition, and safety.

Homologous Overexpression of Tyrosinase in Trichoderma reesei and Its Application in Glycinin Cross-Linking.

Tyrosinase is capable of oxidizing tyrosine residues in proteins, leading to intermolecular protein cross-linking, which could modify the protein network of food and improve the texture of food. To obtain the recombinant tyrosinase with microbial cell factory instead of isolation tyrosinase from the mushroom Agaricus bisporus, a TYR expression cassette was constructed in this study. The expression cassette was electroporated into Trichoderma reesei Rut-C30 and integrated into its genome, resulting in a recombinant strain C30-TYR. After induction with microcrystalline cellulose for 7 days, recombinant tyrosinase could be successfully expressed and secreted by C30-TYR, corresponding to approximately 2.16 g/L tyrosinase in shake-flask cultures. The recombinant TYR was purified by ammonium sulfate precipitation and gel filtration, and the biological activity of purified TYR was 45.6 U/mL. The purified TYR could catalyze the cross-linking of glycinin, and the emulsion stability index of TYR-treated glycinin emulsion was increased by 30.6% compared with the untreated one. The cross-linking of soy glycinin by TYR resulted in altered properties of oil-in-water emulsions compared to emulsions stabilized by native glycinin. Therefore, cross-linking with this recombinant tyrosinase is a feasible approach to improve the properties of protein-stabilized emulsions and gels.

Chicoric acid inserted in protein Z cavity exhibits higher stability and better wound healing effect under oxidative stress.

Oxidative stress is one of the limiting factors that inhibit wound healing. Phytochemicals especially chicoric acid have the potential to act as an antioxidant and scavenge reactive oxygen species, thereby promoting wound healing. However, most of the phytochemicals were easy to be degraded during storage or using due to the oxidative status in wound site. Herein, we introduce a high stable protein Z that can encapsulate chicoric acid during foaming. TEM results showed that the size of protein Z-chicoric acid is in the range of nanoscale (named PZ-CA nanocomposite), and protein Z encapsulation can significantly improve the stability of chicoric acid under oxidative stress. Moreover, PZ-CA nanocomposite exhibited favorable antioxidant properties, biocompatibility, and the ability to promote cell migration in vitro. The role of PZ-CA nanocomposite in skin regeneration was explored by a mice model. Results in vivo suggest that the PZ-CA nanocomposite promotes wound healing with a faster rate as compared with a commercial spray solution, mostly through attenuating the oxidative stress, promoting cell proliferation and collagen deposition. This work not only provides a delivery vector for bioactive molecules, but also develops a kind of nanocomposite with the property of promoting wound healing.

Characterization and Structural Analyses of Enolase from Shrimp (Litopenaeus vannamei).

Transcriptome analysis had recognized enolase from shrimp Litopenaeus vannamei (L. vannamei), which is termed LvEnolase, as one of the allergens, but its amino acid sequence and protein structure have been lacking. In this study, natural LvEnolase was isolated from L. vannamei and characterized for the first time. The full-length cDNA sequence of LvEnolase was effectively cloned, which encoded 434 amino acid residues. The crystal structure of LvEnolase was successfully determined at a resolution of 2.5 Å by X-ray crystallography (PDB: 8UEL). Notably, it was observed that near the active center, a loop exists in either an open or closed state, and the open loop was associated with the product release phase. Furthermore, enzyme activity assays were conducted to validate the catalytic capabilities of purified LvEnolase. These findings significantly enhance our comprehension of the enolase family and provide valuable support for delving into the functions and characteristics of LvEnolase.

Shape-Anisotropic Assembly of Protein Nanocages with Identical Building Blocks by Designed Intermolecular π-π Interactions.

Protein lattices that shift the structure and shape anisotropy in response to environmental cues are closely coupled to potential functionality. However, to design and construct shape-anisotropic protein arrays from the same building blocks in response to different external stimuli remains challenging. Here, by a combination of the multiple, symmetric interaction sites on the outer surface of protein nanocages and the tunable features of phenylalanine-phenylalanine interactions, a protein engineering approach is reported to construct a variety of superstructures with shape anisotropy, including 3D cubic, 2D hexagonal layered, and 1D rod-like crystalline protein nanocage arrays by using one single protein building block. Notably, the assembly of these crystalline protein arrays is reversible, which can be tuned by external stimuli (pH and ionic strength). The anisotropic morphologies of the fabricated macroscopic crystals can be correlated with the Å-to-nm scale protein arrangement details by crystallographic elucidation. These results enhance the understanding of the freedom offered by an object's symmetry and inter-object π-π stacking interactions for protein building blocks to assemble into direction- and shape-anisotropic biomaterials.

Highly Stretchable, Transparent, and Adhesive Double-Network Hydrogel Dressings Tailored with Fish Gelatin and Glycyrrhizic Acid for Wound Healing.

It remains challenging to fabricate highly stretchable and adhesive hydrogel dressings for wound healing using simple, safe, and green methods. Herein, inspired by the main components of snail mucus, a fully physical double-network (DN) hydrogel dressing composed of fish gelatin (FGel) and glycyrrhizic acid (GL) was fabricated, in which FGel provided a protein scaffold to mimic snail mucus proteins, while GL mimicked the adhesion and bioactivity of snail mucus because of its abundant carboxyl and hydroxyl groups and intrinsic immunomodulatory activity. As expected, the obtained FGel/GL hydrogel dressings exhibited outstanding mechanical and adhesive performances (flexibility, stretchability, adhesive ability, and removability), high transparency, and good antifreezing properties. More importantly, they also possessed excellent biocompatibility, cell migration, and angiogenesis ability in vitro experiments. Finally, animal experiments in vivo indicated that the FGel/GL hydrogel dressings significantly promoted full-thickness wound healing, including promoting granulation tissue formation, collagen deposition, and skin angiogenesis and inhibiting the inflammatory response. All these findings indicated that the FGel/GL hydrogel dressings have great potential for applications in the clinical treatment of wound healing.

A Dual Function of Ferritin (Animal and Plant): Its Holo Form for Iron Supplementation and Apo Form for Delivery Systems.

Ferritins represent a class of iron storage proteins with detoxification functions. The importance of these proteins is reflected by their wide distribution throughout the animal and plant kingdoms. Ferritin has two forms: holo and apo. Holo ferritin can act as an efficient and safe factor for iron supplementation, whereas apo ferritin is able to serve as a promising delivery nanovehicle for nutrients and bioactive compounds. So far, the dual functions of ferritins from animal and plant sources have been extensively studied in several fields, such as food, nutrition, medicine, and materials. This review outlines the structure of animal and plant ferritin, the iron supplementation function of holo ferritin, and the delivery function of apo ferritin. Recent advances in iron supplementation and nutrient encapsulation and delivery are highlighted. Finally, the current challenges and future developments for multifunctional applications of ferritins are discussed.

Inhibition of Iron Release from Donkey Spleen Ferritin through Malt-Derived Protein Z-Ferulic Acid Interactions.

Protein-small molecule interactions naturally occur in foodstuffs, which could improve the properties of protein and small molecules. Meanwhile, they might affect the bioavailability and nutritional value of proteins. Ferritin, as an iron-storage protein, has been a focus of research. However, the complexity of foodstuffs enables the interaction between ferritin and food components, especially polyphenols, which can induce iron release from ferritin. Thus, the application of ferritin in food is limited. Inspired by the natural-occurring, strong protein-polyphenol interactions in beer, to inhibit the iron release of ferritin, the malt-derived protein Z (PZ) was chosen to interact with ferulic acid (FA), an abundant reductant in malt, beer, and other foodstuffs. The analysis of the interaction between PZ and FA was carried out using fluorescence spectroscopy, the results of which suggest that one PZ molecule can bind with 22.11 ± 2.13 of FA, and the binding constant is (4.99 ± 2.13) × 105 M-1. In a molecular dynamics (MD) simulation, FA was found to be embedded in the internal hydrophobic pocket of PZ, where it formed hydrogen bonds with Val-389 and Tyr-234. As expected, compared to iron release induced by FA, the iron release from donkey spleen ferritin (DSF) induced by FA decreased by 86.20% in the presence of PZ. Meanwhile, based on the PZ-FA interaction, adding PZ in beer reduced iron release from DSF by 40.5% when DSF:PZ was 1:40 (molar ratio). This work will provide a novel method of inhibiting iron release from ferritin.

Zinc nutrition and dietary zinc supplements.

As the second most abundant trace element in the human body, zinc nutrition is constantly a hot topic. More than one-third population is suffering zinc deficiency, which results in various types of diseases or nutritional deficiencies. Traditional ways of zinc supplementation seem with low absorption rates and significant side effects. Zinc supplements with dietary components are easily accessible and improve zinc utilization rate significantly. Also, mechanisms of maintaining zinc homeostasis are of broad interest. The present review focuses on zinc nutrition in human health in inductive methods. Mainly elaborate on different diseases relating to zinc disorder, highlighting the impact on the immune system and the recent COVID-19. Then raise food-derived zinc-binding compounds, including protein, peptide, polysaccharide, and polyphenol, and also analyze their possibilities to serve as zinc complementary. Finally, illustrate the way to maintain zinc homeostasis and the corresponding mechanisms. The review provides data information for maintaining zinc homeostasis with the food-derived matrix.

Potential enzymes involved in beer monoterpenoids transformation: structures, functions and challenges.

Monoterpenes are important flavor and fragrance compounds in food. In beer, the monoterpenes mainly come from hops added during boiling process. Biotransformations of monoterpene which occurred during fermentation conferred beer with various kinds of aroma profiles, which can be mainly attributed to the contribution of enzymes in yeast. However, there are few reports on the identification and characterization of these enzymes in yeast. Illustrating the structure and functions of key enzymes related to transformations will broaden their potential applications in beer or other foodstuffs. Monoterpenoids including terpene hydrocarbons (limonene, myrcene, and pinene) and terpene alcohol (linalool, geraniol, nerol, and citronellol) gave the beer flower-like or fruit-like aroma. The biotransformation of monoterpenes and monoterpene alcohols in bacteria and yeast, and potential enzymes related to the transformation of them are reviewed here. Enzymes primarily are dehydrogenases including linalool dehydrogenase/isomerase, geraniol/geranial dehydrogenase, nerol dehydrogenase and citronellol dehydrogenase. Most of them are substrate-specific or substrate-specific after modifications by biotechnology methods, and part of them have been expressed in E. coli, while the purification and catalytic rate is very low. Efforts should be made to acquire abundant enzymes, and to fabricate enzyme-expressing yeast, which can be further applied in beer fermentation system.highlightsMonoterpenoids contributed to the flavor of food, especially beer.Transformation of monoterpenoids occurred during fermentation.Various kinds of enzymes are involved in the transformation of monoterpenoids in bacteria, yeast, etc.Crystal structures of these enzymes have been partially resolved.Few enzymes are further applied in food system to obtain abundant flavor.

Response of Saccharomyces cerevisiae var. diastaticus to nerol: Evaluation of antifungal potential by inhibitory effect and proteome analyses.

Saccharomyces cerevisiae var. diastaticus (S. diastaticus) is a major spoilage yeast in brewing. In the present research, the antifungal properties of nerol and the proteome response of S. diastaticus were studied. Results showed nerol can inhibit cell budding and delay yeast fermentation in a dose-depended manner. After 3 d of treatment with 0.25 mg·mL-1 nerol, intracellular ROS levels increased 1.66-fold (P < 0.01), and the cells with damaged membrane increased to 23.2 %. Quantitative proteomic profiles utilizing a capillary-HPLC-MS/MS technology revealed that proteins involved in the metabolism of fermentable sugars were up-regulated in S. diastaticus cells treated with nerol, indicating nerol treatment altered the metabolite pattern of fermentable sugars. Proteins associated with the cell membrane biogenesis, heat shock proteins, amino acid biosynthesis, and glutathione metabolism were similarly up-regulated. These findings revealed the mechanism of nerol-induced yeast cell damage as well as the detoxification response of yeast cells.

Binding of curcumin to barley protein Z improves its solubility, stability and bioavailability.

Although excessive pharmaceutical activities of curcumin have been reported, the poor solubility, low stability and low bioavailability greatly limited its application. In this study, the interaction between protein Z (PZ) and curcumin, and the effects of PZ on the stability and bioavailability of curcumin were investigated. Fluorescence quenching results indicated that curcumin molecule binds PZ with a stoichiometry of 4:1, and the binding affinity is stronger than other reported protein carriers. Molecular dynamics simulation results suggested that curcumin binds in the hydrophobic region of PZ, and the interaction was maintained mainly by hydrogen-bond (Pro-287, Asn-340 and Tyr-234). PZ-curcumin complex possessed better encapsulation efficiency (64.10 %) and loading capacity (5.49 µg/mg) for curcumin. In addition, binding with PZ not only improved the thermal, light and digestive stability of curcumin significantly, but lowered its toxic effect on Caco-2 cells and improved relative bioavailability (305 %) compared with that of curcumin only.

Advances of nanopore-based sensing techniques for contaminants evaluation of food and agricultural products.

Food safety assurance systems are becoming more stringent in response to the growing food safety problems. Rapid, sensitive, and reliable detection technology is a prerequisite for the establishment of food safety assurance systems. Nanopore technology has been taken as one of the emerging technology capable of dealing with the detection of harmful contaminants as efficiently as possible due to the advantage of label-free, high-throughput, amplification-free, and rapid detection features. Start with the history of nanopore techniques, this review introduced the underlying knowledge of detection mechanism of nanopore-based sensing techniques. Meanwhile, sensing interfaces for the construction of nanopore sensors are comprehensively summarized. Moreover, this review covers the current advances of nanopore techniques in the application of food safety screening. Currently, the establishment of nanopore sensing devices is mainly based on the blocking current phenomenon. Sensing interfaces including biological nanopores, solid-state nanopores, DNA origami, and de novo designed nanopores can be used in the manufacture of sensing devices. Food harmful substances, including heavy metals, veterinary drugs, pesticide residues, food toxins, and other harmful substances can be quickly determined by nanopore-based sensors. Moreover, the combination of nanopore techniques with advanced materials has become one of the most effective methods to improve sensing properties.

Recent Advances in the Study of Wheat Protein and Other Food Components Affecting the Gluten Network and the Properties of Noodles.

Upon hydrating and mixing wheat flour, wheat protein forms a network that strongly affects the structure and physicochemical properties of dough, thus affecting the properties of noodles. Different approaches have been taken to alter the gluten network structure in order to control the dough properties. In the current review, we summarize the structure and function of wheat protein, including glutenin and gliadin, and describe food components that may affect noodle quality by interacting with wheat protein. In fact, the ratio of glutenin to gliadin is closely related to the viscosity of dough, and disulfide bonds also contribute to the gluten network formation. Meanwhile, wheat protein coexists with starch and sugar in wheat dough, and thus the nature of starch may highly influence gluten formation as well. Salts, alkali, enzymes and powdered plant food can be added during dough processing to regulate the extensional properties of wheat noodles, obtaining noodles of high quality, with improved sensory and storage properties. This review describes specific methods to reinforce the wheat protein network and provides a reference for improving noodle quality.

Spatiotemporal control over 3D protein nanocage superlattices for the hierarchical encapsulation and release of different cargo molecules.

Taking inspiration from Nature, we have constructed a two-compartment system based on 3D ferritin nanocage superlattices, the self-assembly behavior of which can be spatiotemporally controlled using two designed switches. One pH switch regulates the assembly of the ferritin subunit into its shell-like structure, whereas the other metal switch is responsible for assembly of the 3D superlattices from ferritin nanocages as building blocks. Consequently, this system holds great promise for the hierarchical encapsulation and release of two different cargo molecules.

Directed Self-Assembly of Dimeric Building Blocks into Networklike Protein Origami to Construct Hydrogels.

Engineering proteins to construct self-assemblies is of crucial significance not only for understanding the sophisticated living systems but also for fabricating advanced materials with unexplored functions. However, due to the inherent chemical heterogeneity and structural complexity of the protein surface, designing complex protein assemblies in an anisotropic fashion remains challenging. Here, we describe a self-assembly approach to fabricating protein origami with a networklike structure by designing dual noncovalent interactions on the different positions of a single protein building block. With dimeric proteins as building blocks, 1D protein filaments were constructed by the designed metal coordination at key protein interfaces. Subsequently, the network superstructures were created by the cross-linking of the 1D protein filaments at branch point linkages through the second designed π-π stacking interactions. Notably, upon increasing the protein concentration, the formed protein networks convert into hydrogels with reversible, injectable, and self-healing properties, which have the ability to promote bone regeneration. This strategy could be used to fabricate other protein-based materials with unexplored functions.

Construction of Sol-Gel Phase-Reversible Hydrogels with Tunable Properties with Native Nanofibrous Protein as Building Blocks.

Reversible sol-gel transforming behaviors combined with tunable mechanical properties are vital demands for developing biomaterials. However, it remains challenging to correlate these properties with the hydrogels constructed by denatured protein as building blocks. Herein, taking advantage of naturally high-affinity coordination environments consisting of i, i + 4 His-Glu motifs offered by paramyosin, a ubiquitous nanofibrous protein, we found that Zn2+ rather than Ca2+ or Mg2+ has the ability to trigger the self-assembly of native abalone paramyosin (AbPM) into protein hydrogels under benign conditions, while the addition of EDTA induces the hydrogels back into protein monomers, indicative of a reversible process. By using such sol-gel reversible property, the AbPM gels can serve as a vehicle to encapsulate bioactive molecules such as curcumin, thereby protecting it from degradation from thermal and photo treatment. Notably, based on the high conserved structure of native AbPM, the mechanical property and biological activity of the fabricated AbPM hydrogels can be fined-tuned by its noncovalent interaction with small molecules. All these findings raise the possibility that native paramyosin can be explored as a new class of protein hydrogels which exhibit favorable properties that the traditional hydrogels constructed by denatured protein building blocks do not have.

Fabrication of a donkey spleen ferritin-pectin complex to reduce iron release and enhance the iron supplementation efficacy.

Iron deficiency is a global issue, influencing more than one-third of the population in the world. Ferritin as a natural iron-containing protein is considered a marvelous iron supplement due to its biocompatibility, biodegradability and bioavailability. However, foodstuffs contain plenty of reductants which could induce iron release from the cavity of ferritin and cause oxidative damage. In this study, we aimed to prevent the iron release from donkey spleen ferritin (DSF) by pectin encapsulation driven by the electrostatic interaction and evaluated the iron supplementation of the DSF-pectin complex (DPC). After DSF was purified, we fabricated the DPC and the iron release was decreased by 53.68% after 60 min when DSF : pectin was 1 : 10 (w/w). TEM analysis showed that ferritin in the DPC is clustered in a linear pattern, and the cell viability assay indicated that the DPC has no toxicity towards Caco-2 cells. In the mouse experiment, the DPC increased the content of serum iron and serum ferritin with no significant difference from the control check. Furthermore, the DPC increased the iron content in the liver, suppressed the expression of hepcidin and increased the expression of ferroportin. These results suggested that the DPC could prevent the interactions between food components and ferritin and is a promising iron supplement to ameliorate iron deficiency.

Designing Stacked Assembly of Type III Rubisco for CO2 Fixation with Higher Efficiency.

The slow catalytic rate of the carboxylation enzyme d-ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a major barrier to increasing the rate of carbon assimilation from the atmosphere into the biosphere. It is of great importance to establish a method to improve the carboxylation efficiency of Rubisco. Inspired by the assembly of Rubisco in carboxysomes, herein, we presented a rational protein engineering approach for the construction of one-dimensional (1D) protein arrays of type III Rubisco through designed π-π stacking interactions by using crystal structural information as a guide. In aqueous solutions, the dimensions of these 1D protein arrays collectively span nearly the entire nano- and micrometer scale (200 nm to 5.0 µm) by adjusting protein and NaCl concentrations. As a result, the stacked Rubisco assemblies increase by 40% in the carboxylase activity, while their turnover number (kcat) is around twofold larger than that of wild-type III Rubisco. Notably, upon heat treatment at temperature up to 75 °C for 30 min, most of the assembled nanostructures and the enzyme activity are retained. More importantly, the initial relative activity of stacked assemblies retained 91% after 10 cycles of reuse. This work provides a simple, effective solution for the improvement of the CO2 carboxylation efficiency of Rubisco.