Amelioration of Acute Alcoholic Liver Injury via Attenuating Oxidative Damage and Modulating Inflammation by Means of Ursodeoxycholic Acid-Zein Nanoparticles.

Ursodeoxycholic acid (UDCA) has been broadly adopted for the clinical treatment of hepatic and biliary diseases; however, its poor water-solubility becomes an obstacle in wide applications. To overcome these challenges, herein, a two-tier UDCA-embedded system of zein nanoparticles (NPs) along with a polyelectrolyte complex was designed under facile conditions. Both the UDCA-zein NPs and their inclusion microcapsules showed a spherical shape with a uniform size. A typical wall plus capsule/core structure was formed in which UDCA-zein NPs distributed evenly in the interior. The UDCA inclusion microcapsules had an encapsulation rate of 67% and were released in a non-Fickian or anomalous transport manner. The bioavailability and efficacy of UDCA-zein NPs were assessed in vivo through the alcoholic liver disease (ALD) mouse model via intragastric administration. UDCA-zein NPs ameliorated the symptoms of ALD mice remarkably, which were mainly exerted through attenuation of antioxidant stress levels. Meanwhile, it notably upregulated the intestinal tight junction protein expression and improved and maintained the integrity of the mucosal barrier effectively. Collectively, with the improvement of bioavailability, the UDCA-zein NPs prominently alleviated the oxidative damage induced by alcohol, modulating the inflammation so as to restore ALD. It is anticipated that UDCA-zein NPs have great therapeutic potential as sustained-nanovesicles in ALD treatment.

Synergistic enhancement of glucagon-like peptide-1 release by γ-aminobutyric acid and L-phenylalanine in enteroendocrine cells-searching active ingredients in a water extract of corn zein protein.

This study investigated the glucagon-like peptide-1 (GLP-1)-releasing activity of an aqueous extract (ZeinS) from corn zein protein and aimed to identify the active compounds responsible for this activity. Glucagon-like peptide-1-releasing activity was evaluated using a murine enteroendocrine cell line (GLUTag). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on purified fractions of ZeinS to identify active molecules. ZeinS stimulated more GLP-1 secretion from GLUTag cells compared to zein hydrolysate. Fractions displaying biological activity were determined by solid-phase extraction and high-performance liquid chromatography (HPLC) fractionation. Subsequent LC-MS/MS analysis identified several amino acids in the active fractions of ZeinS. In particular, γ-aminobutyric acid (GABA) exhibited significant GLP-1-releasing activity both alone and synergistically with L-phenylalanine (Phe). Moreover, ZeinS-induced GLP-1 secretion was attenuated by antagonists for the GABA receptor and calcium sensing receptor. These results demonstrate that GABA and Phe identified in ZeinS synergistically stimulate GLP-1 secretion in enteroendocrine cells.

Zein-Based Nanoparticles Improve the Therapeutic Efficacy of a TrkB Agonist toward Alzheimer's Disease.

The brain-derived neurotrophic factor (BDNF)/TrkB pathway plays a crucial role in neural plasticity and neuronal survival but is often deficient in neurodegenerative diseases like Alzheimer's disease (AD). CF3CN acts as a specific TrkB agonist that displays therapeutic effects in the AD mouse model, but its brain/plasma ratio (B/P ratio) distribution is not satisfactory. To increase its brain exposure, we synthesized several derivatives and employed nanoparticle (NP) formulation to optimize the most potent #2 derivative's in vivo PK profiles. We generated stable #2-loaded zein/lactoferrin composite NPs (#2/zein/LF) using the antisolvent co-precipitation method. In vivo PK studies revealed that nanoencapsulation improved #2's oral bioavailability by approximately 2-fold and significantly enhanced its plasma Cmax and t1/2, but the brain profiles were comparable. Pharmacodynamics showed that #2/zein/LF activates TrkB signaling that phosphorylates asparagine endopeptidase (AEP) T322 and decreases its enzymatic activity, resulting in reduced AEP-cleaved amyloid precursor protein and Tau fragments in the brains of AD mice, correlating with its PK profiles. After 3 months of treatment in 3xTg mice, #2/zein/LF decreased AD pathologies and alleviated cognitive dysfunction. Hence, zein/LF composite nanoencapsulation is a promising drug delivery method for improving the PK profiles of a potential preclinical candidate for treating neurodegenerative diseases.

The transcription factors ZmNAC128 and ZmNAC130 coordinate with Opaque2 to promote endosperm filling in maize.

Endosperm filling in maize (Zea mays), which involves nutrient uptake and biosynthesis of storage reserves, largely determines grain yield and quality. However, much remains unclear about the synchronization of these processes. Here, we comprehensively investigated the functions of duplicate NAM, ATAF1/2, and CUC2 (NAC)-type transcription factors, namely, ZmNAC128 and ZmNAC130, in endosperm filling. The gene-edited double mutant zmnac128 zmnac130 exhibits a poorly filled kernel phenotype such that the kernels have an inner cavity. RNA sequencing and protein abundance analysis revealed that the expression of many genes involved in the biosynthesis of zein and starch is reduced in the filling endosperm of zmnac128 zmnac130. Further, DNA affinity purification and sequencing combined with chromatin-immunoprecipitation quantitative PCR and promoter transactivation assays demonstrated that ZmNAC128 and ZmNAC130 are direct regulators of 3 (16-, 27-, and 50-kD) γ-zein genes and 6 important starch metabolism genes (Brittle2 [Bt2], pullulanase-type starch debranching enzyme [Zpu1], granule-bound starch synthase 1 [GBSS1], starch synthase 1 [SS1], starch synthase IIa [SSIIa], and sucrose synthase 1 [Sus1]). ZmNAC128 and ZmNAC130 recognize an additional cis-element in the Opaque2 (O2) promoter to regulate its expression. The triple mutant zmnac128 zmnac130 o2 exhibits extremely poor endosperm filling, which results in more than 70% of kernel weight loss. ZmNAC128 and ZmNAC130 regulate the expression of the transporter genes sugars that will eventually be exported transporter 4c (ZmSWEET4c), sucrose and glucose carrier 1 (ZmSUGCAR1), and yellow stripe-like2 (ZmYSL2) and in turn facilitate nutrient uptake, while O2 plays a supporting role. In conclusion, ZmNAC128 and ZmNAC130 cooperate with O2 to facilitate endosperm filling, which involves nutrient uptake in the basal endosperm transfer layer (BETL) and the synthesis of zeins and starch in the starchy endosperm (SE).

Effects of endosperm type and storage length of whole-plant corn silage on nitrogen fraction, fermentation products, zein profile, and starch digestibility.

Zeins are commercially important proteins found in corn endosperms. The objective of this study was to evaluate the effect of altering zein levels in corn inbred lines carrying endosperm mutations with differential allelic dosage and analyze the effects on the composition, nutritive value, and starch digestibility of whole-plant corn silage (WPCS) at 5 storage lengths. Three inbred lines carrying 3 different endosperm modifiers (opaque-2 [o2], floury-2 [fl2], and soft endosperm-1 [h1]) were pollinated with 2 pollen sources to form pairs of near-isogenic lines with either 2 or 3 doses of the mutant allele for each endosperm modifier. The experiment was designed as a split-plot design with 3 replications. Pollinated genotype was the main plot factor, and storage length was the subplot-level factor. Agronomic precautions were taken to mimic hybrid WPCS to the extent possible. Samples were collected at approximately 30% dry matter (DM) using a forage harvester and ensiled in heat-sealed plastic bags for 0, 30, 60, 120, and 240 d. Thus, the experiment consisted of 30 treatments (6 genotypes × 5 storage lengths) and 90 ensiling units (3 replications per treatment). Measurements included nutrient analysis, including crude protein, soluble crude protein, amylase-treated neutral detergent fiber, acid detergent fiber, lignin, starch, fermentation end products, zein concentration, and in vitro starch digestibility (ivSD). The nutritional profile of the inbred-based silage samples was similar to hybrid values reported in literature. Significant differences were found in fresh (unfermented) sample kernels for endosperm vitreousness and zein profiles between and within isogenic pairs. The o2 homozygous (3 doses of mutant allele) had the highest reduction in vitreousness level (74.5 to 38%) and zein concentration (6.2 to 4.7% of DM) compared with the heterozygous counterpart (2 doses of mutant allele). All genotypes showed significant reduction of total zeins and α-zeins during progressive storage length. In vitro starch digestibility increased with storage length and had significant effects of genotype and storage length but not for genotype by storage length interaction, which suggests that the storage period did not attenuate the difference in ivSD between near-isogenic pairs caused by zeins in WPCS. Both total zeins and α-zeins showed a strong negative correlation with ivSD, which agrees with the general hypothesis that the degradation of zeins increases ruminal starch degradability. Homozygous o2 was the only mutant with significantly higher ivSD compared with the heterozygous version, which suggests that, if all other conditions remain constant in a WPCS systems, substantial reductions in endosperm α-zeins are required to significantly improve ivSD in the silo.

Intestinal Barrier Protective Study of Jujube Peel Polyphenols/Zein Complexes by a Combined Caco-2 Cell and Caenorhabditis elegans Model: A Perspective of Proteomics.

Jujube peels have been recognized as a promising resource of several bioactive ingredients. The main composition of jujube peel polyphenols (JPP) has been identified as rutin, kaempferol-3-O-rutinosid, and salicylic acid. The JPP/zein complexes, whose bioavailability reached 69.73% ± 5.06% in vitro, have been formed successfully. The Caco-2 cell and Caenorhabditis elegans (C. elegans) models have been combined to detect the intestinal barrier protective effect of JPP and its complexes. Results showed that JPP/zein complexes contain better protection capability than JPP in both models. In the Caco-2 cell model, the complex relieved intestinal barrier damage by regulating the tight junction proteins. Moreover, the lysosome pathway has been activated, further regulating immune responses and lipid transportation, improving the barrier function of C. elegans after incubation with JPP/zein complexes according to the proteomics study. This work provides new insights into intestinal barrier protection with bioactive compounds.

Presence of Unabsorbed Free Amino Acids at the End of the Small Intestine Indicates the Potential for an Increase in Amino Acid Uptake in Humans and Pigs.

BACKGROUND: Unabsorbed free amino acids (AAs) at the end of the small intestine result in a potential preventable nutritional loss. OBJECTIVES: This study aimed to quantify free AAs in terminal ileal digesta of both humans and pigs to investigate its relevance for the nutritional value of food proteins. METHODS: Two studies with three diets were performed: a human study-ileal digesta from eight adult ileostomates were collected over 9 h after ingestion of a single meal unsupplemented or supplemented with 30 g zein or whey; pig study-12 cannulated pigs were fed for 7 d with a diet containing whey or zein or no-protein diet, and ileal digesta were collected on the last 2 d. Digesta were analyzed for total and 13 free AAs. True ileal digestibility (TID) of AAs was compared with and without free AAs. RESULTS: All terminal ileal digesta samples contained free AAs. The TID of AAs in whey was 97% ± 2.4% (mean ± SD) in human ileostomates and 97% ± 1.9% in growing pigs. If the analyzed free AAs would have been absorbed, TID of whey would increase by 0.4%-units in humans and 0.1%-units in pigs. The TID of AAs in zein was 70% ± 16.4% in humans and 77% ± 20.6% in pigs and would increase by 2.3%-units and 3.5%-units, respectively, if the analyzed free AAs would have been fully absorbed. The largest difference was observed for threonine from zein: if free threonine was absorbed, the TID would increase by 6.6%-units in both species (P < 0.05). CONCLUSIONS: Free AAs are present at the end of the small intestine and can potentially have a nutritionally relevant effect for poorly digestible protein sources, whereas the effect is negligible for highly digestible protein sources. This result provides insight into the room for improvement of a protein's nutritional value if all free AAs are to be absorbed. J Nutr 2023;xx:xx-xx. This trial was registered at clinicaltrials.gov as NCT04207372.

Nitrogen-dependent binding of the transcription factor PBF1 contributes to the balance of protein and carbohydrate storage in maize endosperm.

Fluctuations in nitrogen (N) availability influence protein and starch levels in maize (Zea mays) seeds, yet the underlying mechanism is not well understood. Here, we report that N limitation impacted the expression of many key genes in N and carbon (C) metabolism in the developing endosperm of maize. Notably, the promoter regions of those genes were enriched for P-box sequences, the binding motif of the transcription factor prolamin-box binding factor 1 (PBF1). Loss of PBF1 altered accumulation of starch and proteins in endosperm. Under different N conditions, PBF1 protein levels remained stable but PBF1 bound different sets of target genes, especially genes related to the biosynthesis and accumulation of N and C storage products. Upon N-starvation, the absence of PBF1 from the promoters of some zein genes coincided with their reduced expression, suggesting that PBF1 promotes zein accumulation in the endosperm. In addition, PBF1 repressed the expression of sugary1 (Su1) and starch branching enzyme 2b (Sbe2b) under normal N supply, suggesting that, under N-deficiency, PBF1 redirects the flow of C skeletons for zein toward the formation of C compounds. Overall, our study demonstrates that PBF1 modulates C and N metabolism during endosperm development in an N-dependent manner.

Electrospun Mucoadhesive Zein/PVP Fibroporous Membrane for Transepithelial Delivery of Propranolol Hydrochloride.

Mucoadhesive drug delivery systems have been extensively studied to effectively reduce the limitations of conventional drug delivery systems. Zein and polyvinyl pyrrolidone (PVP) are appraised for mucoadhesive properties. This study focuses on developing a mechanically stable zein/PVP electrospun membrane for propranolol hydrochloride (PL) transport. Fourier transform infrared, Raman spectra, and swelling studies gave evidence for PVP crosslinking, whereas circular dichroism spectroscopy revealed crosslinking of zein owing to the conformational change from α-helix to ß-sheet. A 10 h thermal treatment of zein/PVP imparted 3.92 ± 0.13 MPa tensile strength to the matrix. Thermally crosslinked electrospun zein/PVP matrix showed 22.1 ± 0.1 g mm work of adhesion in porcine buccal mucosa tissue. Qualitative and quantitative evaluation of cytotoxicity in RPMI 2650 has been carried out. The in vitro drug release profile of PL from thermally crosslinked zein/PVP best fitted with the Korsmeyer-Peppas model. Immunostaining of ß-catenin adherens junctional protein confirmed the absence of paracellular transport through the junctional opening. Still, drug permeation was observed through the porcine buccal mucosa, attributed to the transcellular transport of PL owing to its lipophilicity. The ex vivo permeation of PL through porcine buccal mucosa was also evaluated.

Effect of multi-mode sweep frequency ultrasound pretreatment on properties of the zeins and ACE inhibitory peptides activity of the hydrolysates.

Effects of sweep frequency ultrasound (SFU) pretreatment of a new multifunctional ultrasonic equipment on hydrolysis characteristics of zeins and angiotensin-converting enzyme (ACE) inhibitory activity of zein hydrolysates were investigated. Degree of hydrolysis of zeins reached the highest of 25.93 % and 25.72 % at 40 kHz and 25/40 kHz, respectively. While 25/40 kHz increased solubility, surface hydrophobicity, particle size uniform of zeins and ACE inhibitory activities of the hydrolysates significantly. Endogenous fluorescence indicated that 25/40 kHz promoted unfolding of protein molecules and exposure of hydrophobic residues, thereby facilitating enzymatic hydrolysis. Circular dichroism spectrum and Fourier transform infrared spectrometer illustrated that 25/40 kHz unfolded protein molecules and decreased α-helical contents remarkably. Gel permeation chromatography showed that more small-molecule active peptides were obtained from hydrolysates at 25/40 kHz. In conclusion, SFU pretreatment at 25/40 kHz with the new equipment before proteolysis is an efficient method to improve ACE inhibitory activity of the hydrolysates.

Endosperm-specific accumulation of human α-lactalbumin increases seed lysine content in maize.

KEY MESSAGE: This study demonstrated high expression and accumulation of human α-lactalbumin in transgenic maize, and significant improvement of lysine content in maize endosperm. As a high-yield crop, lack of lysine in endosperm storage protein is a major defect of maize (Zea mays L.). Specifically expression of foreign proteins is a potential way to improve lysine content in maize endosperm. Human α-lactalbumin is such a protein with high lysine content and high nutritional value. In this study, the codon-optimized human lactalbumin alpha (LALBA) gene was driven by maize endosperm-specific 27 kD γ-zein promoter, and transformed into maize. Five independent transgenic lines were obtained, and LALBA was highly expressed in endosperm in all these lines. Protein assay indicated that human α-lactalbumin was highly accumulated in maize endosperm. Immuno-localization assay indicated that human α-lactalbumin was mainly deposited into the protein body (PB). Protein interaction assay showed that human α-lactalbumin interacted with 16 kD γ-zein, which might lead to its deposition to the PBs. Amino acid analysis of two independent transgenic lines showed significant increase of lysine contents in transgenic endosperm, with 47.26% and 45.15% increase to their non-transgenic seeds, respectively. We obtained transgenic maize with endosperm-specific accumulation of human α-lactalbumin at high level and increased the lysine content in maize endosperm. This study demonstrated an effective way to improve the nutritional value of maize seeds.

Zein enhanced the digestive stability of five citrus flavonoids via different binding interaction.

BACKGROUND: Zein is commonly used to construct food flavonoid delivery systems. This study investigated the effect and mechanism of zein on the digestive stability of five citrus flavonoids, namely hesperetin (HET), hesperidin (HED), neohesperidin (NHD), naringenin (NEN), and naringin (NIN). RESULTS: Zein enhanced the digestive stability of the five citrus flavonoids, especially that of HET and NEN, during digestion in the stomach and small intestine. Fluorescence spectroscopy results suggested that citrus flavonoids spontaneously quenched the endogenous fluorescence of zein in static quenching mode. The binding of HET, HED and NHD to zein was driven respectively by electrostatic, hydrophobic and electrostatic interaction. However, Van der Waals' force and hydrogen (H)-bond interaction represented the primary driving force for binding NEN, and NIN to zein to form complexes. The binding of the five citrus flavonoids to zein also caused a diverse bathochromic shift in ultraviolet absorbance. Analysis using Fourier-transform infrared and Raman spectroscopy revealed that the binding behavior of the five citrus flavonoids had different effects on changes in the secondary structures, disulfide bonds, and tyrosine exposure of zein. The results were also partially verified by molecular dynamic simulation. CONCLUSIONS: Zein enhanced the digestive stability of the five citrus flavonoids via different binding interactions that was due to the difference in molecular structure of citrus flavonoids. © 2022 Society of Chemical Industry.

Progressive Aggregation of 16 kDa Gamma-Zein during Seed Maturation in Transgenic Arabidopsis thaliana.

Prolamins constitute a unique class of seed storage proteins, present only in grasses. In the lumen of the endoplasmic reticulum (ER), prolamins form large, insoluble heteropolymers termed protein bodies (PB). In transgenic Arabidopsis (Arabidopsis thaliana) leaves, the major maize (Zea mays) prolamin, 27 kDa γ-zein (27γz), assembles into insoluble disulfide-linked polymers, as in maize endosperm, forming homotypic PB. The 16 kDa γ-zein (16γz), evolved from 27γz, instead forms disulfide-bonded dispersed electron-dense threads that enlarge the ER lumen without assembling into PB. We have investigated whether the peculiar features of 16γz are also maintained during transgenic seed development. We show that 16γz progressively changes its electron microscopy appearance during transgenic Arabidopsis embryo maturation, from dispersed threads to PB-like, compact structures. In mature seeds, 16γz and 27γz PBs appear very similar. However, when mature embryos are treated with a reducing agent, 27γz is fully solubilized, as expected, whereas 16γz remains largely insoluble also in reducing conditions and drives insolubilization of the ER chaperone BiP. These results indicate that 16γz expressed in the absence of the other zein partners forms aggregates in a storage tissue, strongly supporting the view that 16γz behaves as the unassembled subunit of a large heteropolymer, the PB, and could have evolved successfully only following the emergence of the much more structurally self-sufficient 27γz.

ZmGRAS11, transactivated by Opaque2, positively regulates kernel size in maize.

Although the genetic basis for endosperm development in maize (Zea mays) has been well studied, the mechanism for coordinating grain filling with increasing kernel size remains elusive. Here, we report that increased kernel size was selected during modern breeding and identify a novel DELLA-like transcriptional regulator, ZmGRAS11, which positively regulates kernel size and kernel weight in maize. We find that Opaque2, a core transcription factor for zein protein and starch accumulation, transactivates the expression of ZmGRAS11. Our data suggest that the Opaque2-ZmGRAS11 module mediates synergistic endosperm enlargement with grain filling.

Two γ-zeins induce the unfolded protein response.

The rapid, massive synthesis of storage proteins that occurs during seed development stresses endoplasmic reticulum (ER) homeostasis, which activates the ER unfolded protein response (UPR). However, how different storage proteins contribute to UPR is not clear. We analyzed vegetative tissues of transgenic Arabidopsis (Arabidopsis thaliana) plants constitutively expressing the common bean (Phaseolus vulgaris) soluble vacuolar storage protein PHASEOLIN (PHSL) or maize (Zea mays) prolamins (27-kDa γ-zein or 16-kDa γ-zein) that participate in forming insoluble protein bodies in the ER. We show that 16-kDa γ-zein significantly activates the INOSITOL REQUIRING ENZYME1/BASIC LEUCINE ZIPPER 60 (bZIP60) UPR branch-but not the bZIP28 branch or autophagy-leading to induction of major UPR-controlled genes that encode folding helpers that function inside the ER. Protein blot analysis of IMMUNOGLOBULIN-BINDING PROTEIN (BIP) 1 and 2, BIP3, GLUCOSE REGULATED PROTEIN 94 (GRP94), and ER-localized DNAJ family 3A (ERDJ3A) polypeptides confirmed their higher accumulation in the plant expressing 16-kDa γ-zein. Expression of 27-kDa γ-zein significantly induced only BIP3 and ERDJ3A transcription even though an increase in GRP94 and BIP1/2 polypeptides also occurred in this plant. These results indicate a significant but weaker effect of 27-kDa γ-zein compared to 16-kDa γ-zein, which corresponds with the higher availability of 16-kDa γ-zein for BIP binding, and indicates subtle protein-specific modulations of plant UPR. None of the analyzed genes was significantly induced by PHSL or by a mutated, soluble form of 27-kDa γ-zein that traffics along the secretory pathway. Such variability in UPR induction may have influenced the evolution of storage proteins with different tissue and subcellular localization.

Enzyme- and Relative Humidity-Responsive Antimicrobial Fibers for Active Food Packaging.

Active food packaging materials that are sustainable, biodegradable, and capable of precise delivery of antimicrobial active ingredients (AIs) are in high demand. Here, we report the development of novel enzyme- and relative humidity (RH)-responsive antimicrobial fibers with an average diameter of 225 ± 50 nm, which can be deposited as a functional layer for packaging materials. Cellulose nanocrystals (CNCs), zein (protein), and starch were electrospun to form multistimuli-responsive fibers that incorporated a cocktail of both free nature-derived antimicrobials such as thyme oil, citric acid, and nisin and cyclodextrin-inclusion complexes (CD-ICs) of thyme oil, sorbic acid, and nisin. The multistimuli-responsive fibers were designed to release the free AIs and CD-ICs of AIs in response to enzyme and RH triggers, respectively. Enzyme-responsive release of free AIs is achieved due to the degradation of selected polymers, forming the backbone of the fibers. For instance, protease enzyme can degrade zein polymer, further accelerating the release of AIs from the fibers. Similarly, RH-responsive release is obtained due to the unique chemical nature of CD-ICs, enabling the release of AIs from the cavity at high RH. The successful synthesis of CD-ICs of AIs and incorporation of antimicrobials in the structure of the multistimuli-responsive fibers were confirmed by X-ray diffraction and Fourier transform infrared spectrometry. Fibers were capable of releasing free AIs when triggered by microorganism-exudated enzymes in a dose-dependent manner and releasing CD-IC form of AIs in response to high relative humidity (95% RH). With 24 h of exposure, stimuli-responsive fibers significantly reduced the populations of foodborne pathogenic bacterial surrogates Escherichia coli (by ∼5 log unit) and Listeria innocua (by ∼5 log unit), as well as fungi Aspergillus fumigatus (by >1 log unit). More importantly, the fibers released more AIs at 95% RH than at 50% RH, which resulted in a higher population reduction of E. coli at 95% RH. Such biodegradable, nontoxic, and multistimuli-responsive antimicrobial fibers have great potential for broad applications as active and smart packaging systems.

Carbonized zein nanosheets with intrinsic enzyme-mimicking activities and high photothermal conversion efficiency for synergistic cancer therapy.

With the rapid development of biology and nanotechnology, designing nanomaterials with intrinsic enzyme-like activities has attracted huge attention in recent years. Herein, for the first time, we use zein as a new protein precursor to prepare N-rich carbonized zein nanosheets (C-Zein) via facile pyrolysis. Zein is an inert, biodegradable and sustainable natural biopolymer. After high-temperature carbonization, zein can be converted into highly catalytically active C-Zein, which can possess excellent peroxidase- and oxidase-like catalytic activities. Such intrinsic enzyme-like activities of C-Zein are closely related to its graphitization degree, the ratio of graphitic nitrogen and the formation of disordered graphene. Intriguingly, C-Zein also exhibits high photothermal conversion efficiency in the near-infrared (NIR) region. Coupling their unique photothermal and catalytic properties, the as-prepared C-Zein can act as a robust agent for synergistic photothermal-catalytic cancer treatment under the irradiation of NIR light. We expect that this work paves the way to use zein for designing efficient artificial enzymes and accelerate further growth in exploring its new biomedical and pharmaceutical applications.

Effect of temperature and pH on the conversion between free and hidden zearalenone in zein.

Food processing might induce the transformation of hidden ZEN (zein-bound ZEN) in maize. The objective of this study was to assess the effect of processing factors on free ZEN and hidden ZEN. After zein was treated under different temperature and pH, ZEN was quantified in samples before and after in vitro digestion. The ratios of hidden to total ZEN in zein are decreased from 54.25% to 40.74% after thermal treatment and from 54.25% to 0 after alkaline treatment, respectively. Conversely, acid treatment increased the ratio of hidden to total ZEN from 54.25% to 100%. Thus, it can be concluded that thermal or alkaline condition induced the conversion of hidden ZEN to free ZEN while acid condition promoted the ZEN-zein interactions to form the hidden ZEN. Overall, temperature and pH values played a vital role in the conversion of hidden ZEN during food processing.

Integrated multi-spectroscopic and molecular modeling techniques to study the formation mechanism of hidden zearalenone in maize.

Hidden mycotoxins have been reported to be "protected" by macromolecular substances to escape routine determination, but release to free mycotoxins under gastrointestinal conditions. Nowadays, the hidden zearalenone (ZEN) that binding with macromolecular zein has been found in maize. However, the binding mechanism of ZEN with zein in maize has not been clarified. In this study, the formation of ZEN-zein complex was investigated applying ultrafiltration, multi-spectroscopic and molecular modeling techniques. The steady-state and transient fluorescence analysis suggested the ZEN could interact with zein to form the complex driven by hydrophobic force and hydrogen bonds, which is in accordance with the molecular modeling studies. The conformational changes of zein induced by binding with ZEN were revealed by Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD). Elucidating the binding mechanism between zein and ZEN could help the development of detecting hidden ZEN and guarantee the safety of maize products.

Tuning of Molecular Interactions between Zein and Tannic Acid to Modify Sunflower Sporopollenin Exine Capsules: Enhanced Stability and Targeted Delivery of Bioactive Macromolecules.

There are multiple obstacles for the storage and digestion of orally administered bioactive macromolecules. This study developed a low-cost and sustained-release delivery system (sporopollenin exine capsules with zein/tannic acid modification) of proteins with excellent storage stability, and at the same time provided insights into the sustained-release mechanism through exploring the interaction between zein and tannic acid (TA). ß-Galactosidase (ß-Gal) was utilized as a model protein and loaded into sporopollenin exine capsules (SECs), which were then coated with the zein/TA system. Under the optimized zein/TA conditions, the zein/TA system showed better performance than the zein alone system in the sustained release of ß-Gal, with the residual activity of about 70.26% after 24 h of simulated digestion. Evaluation of the storage stability demonstrated a ß-Gal residual activity of nearly 90% for 28 days at 25 °C. Additionally, FTIR analysis demonstrated that the stability of the zein/TA system depends on both hydrogen bonding and certain covalent bonding through the Schiff-base reaction, and the sustained release is regulated by the bonding strength.