Biosynthesis of the phycocyanin ß-subunit in Escherichia coli BL21 and its antioxidant activity and application in the preservation of fresh-cut apples.

In this study, the biosynthesis of phycocyanin ß-subunit (CpcB) in Escherichia coli BL21 was investigated, and its antioxidant activity and application in anti-browning of fresh-cut apples was explored. Four genes (cpcB, cpeS, hox1 and pcyA) involved in the biosynthesis of CpcB were cloned and transformed into E. coli BL21 by constructing recombinant plasmid pETDuet-5. The positive transformant was screened by ampicillin resistance. The analysis of SDS-PAGE and zinc fluorescence spectrum showed that CpcB was successfully expressed in E. coli BL21 with a molecular weight of 21 kDa. The purified CpcB had a maximum absorption peak at 615 nm, and its maximum florescence emission wavelength was 640 nm. It exhibited a stronger ability to scavenge four free radicals than Vc. The color change in fresh-cut apples was obviously delayed by the CpcB treatment. These results suggest that CpcB may be used as a potential anti-browning agent for food preservation.

High intestinal isoleucine is a potential risk factor for food allergy by regulating the mTOR/AKT pathway in dendritic cells.

Food allergy is a global food safety problem with a growing prevalence. People in industrial regions are more susceptible to allergy, but the mechanisms behind this are not fully understood. In this study, the probiotic Lactobacillus casei Zhang (LcZ) was administered to allergic individuals and the impact on allergy-related factors were determined. LcZ alleviated allergenic responses, and there was a significant correlation between the intestinal isoleucine content and IgE concentration. Metagenomics results suggest that the metabolism of the gut microbiota is a source of isoleucine. In a mouse model of food allergy, a high isoleucine diet exacerbated allergic responses and increased the activity of allergenic dendritic cell. In a dendritic cell model, a protein array revealed that the mTOR/AKT pathway mediated the function of isoleucine, and molecular docking suggested that Sestrin2 could be the potential receptor. Overall, this study revealed the role of isoleucine in promoting food allergy, elucidated the underlying mechanisms, and suggested that a high intake of isoleucine could be a potential risk factor for food allergy.

Effect of glycation derived from α-dicarbonyl compounds on the in vitro digestibility of ovalbumin: Tracing of advanced glycation end-products and immuno-active peptides.

Currently, the biological consequences of advanced glycation end-products (AGEs) and their link to the antigenicity of food allergens are largely unknown due to the uncertainty in their digestive fates within the body. In this study, the influence of glycation derived from α-dicarbonyl compounds (α-DCs), precursors of AGEs, on digestive behaviors of ovalbumin (OVA) was investigated in a two-step simulated gastrointestinal (GI) model. Methylglyoxal (MGO), glyoxal (GO), and 3-deoxyglucosone were selected as typical α-DCs to obtain glycated OVA with different AGE-modifications (AGE-Ms). It was unveiled that α-DC-glycation reduced the digestibility of OVA via blocking tryptic cleavage sites and inducing steric hindrance, especially seen in the GO- and MGO-OVA groups. The formed AGE-Ms, depending on the precursor type, showed masking effects on the epitopes of OVA, which counteracted the negative effects of reduced digestibility on its antigenicity. Substantial changes in the peptide release patterns were also noted in glycated OVA, including alterations in the sequences and structures of several known protease-resistant epitopes of OVA. This study provides new insights into the nutritional and healthy effects of MRPs in heat-processed foods, as well as their potential connection to the modulation of egg allergy.

Parallel generation of extra advanced glycation end-products during co-digestion of whey proteins and α-dicarbonyls in a simulated gastrointestinal model.

Advanced glycation end-products (AGEs) are a group of heterogeneous compounds formed during the Maillard Reaction (MR) and have been proven to be detrimental to human health. In addition to thermally processed foods, the digestive tract may be an additional site for exogenous AGE formation since the MR would possibly occur between (oligo-)peptides, free amino acids, and reactive MR products (MRPs) such as α-dicarbonyl compounds (α-DCs) along the digestion. In this study, through establishing a simulated gastrointestinal (GI) model consisting of whey protein isolate (WPI) and two typical α-DCs, i.e., methylglyoxal (MGO) or glyoxal (GO), we first validated that co-digestion of WPI with α-DCs generated extra amounts of AGEs in a precursor-dependent manner, especially seen in the intestinal stage. At the end of GI digestion, the contents of total AGEs in WPI-MGO and WPI-GO systems were 4.3-242 and 2.5-73.6 times higher than those formed in the control system, respectively. Evaluation of the protein digestibility further showed that AGE formation along the digestion process slightly affected the digestibility of whey protein fractions. However, as sequenced and identified by high-resolution mass spectrometry, different types of AGE modifications were identified in peptides released from ß-lactoglobulin and α-lactalbumin in the final digests, as well as changes in peptide sequence motifs. This suggested that the glycated structures formed during co-digestion affected the action of digestive proteases toward whey proteins. Overall, these results highlight the GI tract as an additional source of exogenous AGEs and provide new insights into the biochemical consequences of MRPs in heat-processed foods.

Extraction, Structural Characterization, Biological Functions, and Application of Rice Bran Polysaccharides: A Review.

Rice bran is a "treasure house of natural nutrition". Even so, utilization of rice bran is often ignored, and this has resulted in the wastage of nutrients. Polysaccharides are one of the active substances in rice bran that have gained widespread attention for their antioxidant, antitumor, immune-enhancing, antibacterial, and hypoglycemic properties. This review summarizes the extraction methods, structural characterization, bioactivity, and application of rice bran polysaccharides that have been developed and studied in recent years, laying a foundation for its development into foods and medicines. In addition, we also discuss the prospects for future research on rice bran polysaccharides.

Identification of allergens and allergen hydrolysates by proteomics and metabolomics: A comparative study of natural and enzymolytic bee pollen.

Bee pollen as a plant-derived food is consumed as nutritional/functional supplements by humans. But it might confer foodborne allergenicity in susceptible populations, limiting its extensive application. In this study, five potential allergens including profilin, cystatin, prolamin, expansin, and alcohol dehydrogenase in bee pollen derived from Brassica campestris (BP-Bc), were identified through mass spectrometry-based proteomic analysis. Moreover, different types of enzymes (cellulases, pectases, and papains) serve biological roles in pollen wall breaking and expansion, but also promote allergen release and degradation. Proteomic analysis showed that profilin, cystatin, and alcohol dehydrogenase were significantly reduced in BP-Bc following joint treatment with three enzymes. Metabolomic characterization of potential enzymatic hydrolysates of these significantly-decreased allergens was performed, which showed nine major oligopeptides and six amino acids at significantly higher levels in the enzyme-treated BP-Bc. These findings clarified the culprit responsible for bee pollen allergy and the mechanism of enzymatic desensitization for its further development.

Antibacterial Gelidium amansii polysaccharide-based edible films containing cyclic adenosine monophosphate for bioactive packaging.

A homogeneous polysaccharide (GAP), with a molecular weight of 51.8 kDa, was isolated from edible red seaweed Gelidium amansii. Composition analysis suggested GAP contained 5.31% sulfate and 17.33% 3,6-anhydro-galactose and was mainly composed of galactose. Furthermore, GAP, as a biopolymer matrix, was used to form the composite films with the small biological molecules cytidine-5'-monophosphate (CMP), adenosine-5'-monophosphate (AMP), and cyclic adenosine monophosphate (cAMP). Scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectrum, and X-ray diffraction (XRD) results showed that CMP, AMP, and cAMP interacted with the film substrates and might made films more complex. Notably, the addition of CMP, AMP, and cAMP promoted the light, water vapor, and oxygen barrier ability, surface wettability, mechanical strength, and antimicrobial activity against Gram-negative and -positive bacteria. Finally, GAP-based films composited with cAMP (cAMPF) exhibited the best characteristics were applied to fish packaging and preservation at 4 °C and extended the fish shelf life. All these data suggested the potential value of cAMPF as a functional edible polysaccharide film applied in food industries.

Degraded polysaccharides from Porphyra haitanensis: purification, physico-chemical properties, antioxidant and immunomodulatory activities.

To explore effect of the structural properties of porphyra haitanensis polysaccharide on its biological activity, degraded porphyra polysaccharides were separated and purified by Cellulose DEAE-52 and Sephadex G-100 chromatography, obtaining three purified components (P1, P2 and P3). All the three components were sulfate polysaccharides containing the repeating units of → 3) ß-D-galactose (1 → 4) 3,6-anhydro-α-L-galactose (1 →, and → 3) ß-D-galactose (1 → 4) α-L-galactose-6-S (1 →, and → 3) 6-O-methyl-ß-D-galactose (1 → 4) 3,6-anhydro-α-L-galactose (1 →. The molecular weight of the three fractions was measured to be 300.3, 130.4 and 115.1 kDa, respectively. Their antioxidant activity was investigated by the determination of the free radical scavenging effect and ferric reducing power. It was found that P1, P2 and P3 possessed marked antioxidant activity. It was also found that they appreciably enhanced the proliferation, phagocytic ability and nitric oxide secretion in RAW264.7 cells. Lower molecular weight and higher sulfate content were beneficial to bioactivities of P. haitanensis polysaccharides. Overall, P2 and P3 possess superior immuno-modulatory activity to that of P1 and PHP. Thus, the current work will provide the basis for the better utilization of P. haitanensis to develop the related functional foods.

Suppression of Hippo Pathway by Food Allergen Exacerbates Intestinal Epithelia Instability and Facilitates Hypersensitivity.

SCOPE: Hippo signaling is a crucial pathway in innate immune responses, but the relationship between food allergy and Hippo pathway is unknown. The aim of this work is to investigate the regulation of food allergy by Hippo pathway and reveal the molecular mechanisms. METHODS AND RESULTS: Two food allergens tropomyosin and ovalbumin are used to challenge a mouse model and CMT93 intestinal epithelia cell model. The allergic responses and the activation of Hippo pathway are tested in these models. In the mouse model, both allergens trigged significant allergic responses, and Hippo pathway is suppressed after allergen challenge. In CMT93, both allergens upregulate the expression of allergic cytokines thymic stromal lymphopoietin, interleukin (IL)-25, and IL-33. In TAZ KD CMT93, the Hippo pathway is blocked, and the expression of allergenic cytokines are also suppressed. CONCLUSIONS: Both in vivo and in vitro data demonstrate that the two food allergens suppressed Hippo pathway by downregulating TAZ expression, resulting in intestinal epithelia instability, and finally leading to hypersensitivity reactions. These findings provide potential therapeutic targets and molecular markers for food allergy, and provide dietary guidelines for allergenic individuals.

Improved antioxidant and immunomodulatory activities of enzymatically degraded Porphyra haitanensis polysaccharides.

Porphyra haitanensis polysaccharide (CPH) was degraded by pectinase to improve its biological activities. Box-Behnken response surface design was used to optimize the hydrolysis conditions. The molecular weight of CPH and the degraded P. haitanensis polysaccharide (DCPH) were measured to be 524 and 217 kDa, respectively. GC-MS spectrometry results showed that CPH and DCPH were mainly composed of galactose. In vitro antioxidant assays indicated that DCPH possessed improved radical scavenging activity and ferric iron reducing power when compared to those of CPH. In H2 O2 -treated RAW264.7 cells, DCPH was also found to be more effective in reducing the generation of malondialdehyde and reactive oxygen species than CPH. The immunomodulatory assays demonstrated that DCPH possessed superior activities in enhancing the proliferation, phagocytosis, and NO secretion in a RAW264.7 macrophage cell model to those of CPH. PRACTICAL APPLICATIONS: Polysaccharide is the most abundant bioactive component of an edible red algae Porphyra haitanensis. However, the use of CPH is limited due to its relatively low biological activities. Thus, in order to fully utilize P. haitanensis, it is necessary to enhance the biological activities of CPH for its practical use. An efficient and practical method to enhance the bioactivities of P. haitanensis polysaccharide has been developed in the present work. The DCPH prepared in this work could have potential applications in food and medicinal areas.

Two polysaccharides from Porphyra modulate immune homeostasis by NF-κB-dependent immunocyte differentiation.

Porphyra polysaccharides possess multiple pharmacological activities, such as immunoregulatory, anti-tumor and anti-inflammatory effects, but the specific underlying mechanisms are not fully understood. The present work was to investigate the immunomodulatory activity of two different Porphyra polysaccharides, PH and PY, in a BALB/c mouse model and a mouse splenic cell model. Results showed that PH and PY regulated Th1 and Th2 responses, which could be due to the proliferation of CD4+CD25+ Treg. Further investigations demonstrated that PY induced the proliferation and maturation of upstream MHC II+CD11c+ DC. Moreover, both PH and PY activate NF-κB signaling pathways in splenic cells, but the loss-of-function assay with a NF-κB inhibitor demonstrated that the direct Treg-induction activity of PH, but not PY, was mediated by NF-κB. These results suggested that PH and PY show strong immunomodulatory activity by NF-κB-dependent immunocyte maturation and differentiation, which facilitates further application of Porphyra polysaccharides as potential functional foods or immunomodulators.

Quorum sensing system-regulated genes affect the spoilage potential of Shewanella baltica.

Large yellow croaker (Pseudosciaena crocea) is a popular and nutritious but also highly perishable fish species, with Shewanella baltica being the primary spoilage bacteria during low-temperature storage. Clarifying the factors promoting spoilage will facilitate efforts to predict and control the shelf life of foods. This study focused on spoilage-related genes in two Shewanella baltica strains with different spoilage potentials. Using whole genome sequencing and alignment, three distinguishing genes (torT, cysM and trxB) were identified. Further protein sequence comparison and protein structure modeling revealed possible motifs responsible for the spoilage activity. Moreover, diketopiperazine (DKP) quorum sensing (QS) signaling molecules regulated biofilm formation and spoilage gene expression, indicating a relationship between the QS system, biofilm formation and spoilage potential. Our results suggest that DKPs and spoilage genes are potential targets for developing novel food antiseptics, as well as new markers for fish product spoilage.

Intestinal inflammation modulates expression of the iron-regulating hormone hepcidin depending on erythropoietic activity and the commensal microbiota.

States of chronic inflammation such as inflammatory bowel disease are often associated with dysregulated iron metabolism and the consequent development of an anemia that is caused by maldistribution of iron. Abnormally elevated expression of the hormone hepcidin, the central regulator of systemic iron homeostasis, has been implicated in these abnormalities. However, the mechanisms that regulate hepcidin expression in conditions such as inflammatory bowel disease are not completely understood. To clarify this issue, we studied hepcidin expression in mouse models of colitis. We found that dextran sulfate sodium-induced colitis inhibited hepcidin expression in wild-type mice but upregulated it in IL-10-deficient animals. We identified two mechanisms contributing to this difference. Firstly, erythropoietic activity, as indicated by serum erythropoietin concentrations and splenic erythropoiesis, was higher in the wild-type mice, and pharmacologic inhibition of erythropoiesis prevented colitis-associated hepcidin downregulation in these animals. Secondly, the IL-10 knockout mice had higher expression of multiple inflammatory genes in the liver, including several controlled by STAT3, a key regulator of hepcidin. The results of cohousing and fecal transplantation experiments indicated that the microbiota was involved in modulating the expression of hepcidin and other STAT3-dependent hepatic genes in the context of intestinal inflammation. Our observations thus demonstrate the importance of erythropoietic activity and the microbiota in influencing hepcidin expression during colitis and provide insight into the dysregulated iron homeostasis seen in inflammatory diseases.

Effect of selenium nanoparticles with different sizes in primary cultured intestinal epithelial cells of crucian carp, Carassius auratus gibelio.

Nano-selenium (Se), with its high bioavailability and low toxicity, has attracted wide attention for its potential application in the prevention of oxidative damage in animal tissues. However, the effect of nano-Se of different sizes on the intestinal epithelial cells of the crucian carp (Carassius auratus gibelio) is poorly understood. Our study showed that different sizes and doses of nano-Se have varied effects on the cellular protein contents and the enzyme activities of secreted lactate dehydrogenase, intracellular sodium potassium adenosine triphosphatase, glutathione peroxidase, and superoxide dismutase. It was also indicated that nano-Se had a size-dependent effect on the primary intestinal epithelial cells of the crucian carp. Thus, these findings may bring us a step closer to understanding the size effect and the bioavailability of nano-Se on the intestinal tract of the crucian carp.

Effect of tea polyphenols on microbiological and biochemical quality of Collichthys fish ball.

BACKGROUND: Tea polyphenols (TP), as the most active constituents of tea, are considered natural food additives. This study examined the preservative properties of TP for Collichthys fish ball in well storage. Vacuum-packed Collichthys fish balls were treated with 0, 0.1, 0.15, 0.20, 0.25, and 0.30 g kg(-1) TP and stored at 0 °C for 17 days. RESULTS: Microbiological results were obtained using a biochemical test, API system kit, and 16S rDNA sequence analysis. Results confirmed that the dominant bacteria in Collichthys fish balls are the genera Serratia and Pseudomonas. Total viable counts dropped two orders of magnitude in Collichthys fish balls with 0.25 g kg(-1) TP compared with the control. The advantages of total volatile basic nitrogen value, 2-thiobarbituric acid value and texture value were clearly observed, whereas pH and whiteness value exhibited no significant decrease for the group treated with 0.25 g kg(-1) TP. More than 0.25 g kg(-1) TP added could retain excellent fish ball characteristics in terms of sensory assessment after 17 days. CONCLUSION: The shelf life of Collichthys fish balls supplemented with tea polyphenols can be prolonged for an additional 6 days in good condition at 0 °C storage.

Tea polyphenols inhibit Pseudomonas aeruginosa through damage to the cell membrane.

This paper aims to delineate the inhibition mechanism of tea polyphenols (TP) toward Pseudomonas aeruginosa by cell membrane damage. Morphological changes in bacteria treated with TP were investigated by transmission electron microscopy, with results indicating that the primary inhibitory action of TP is to damage bacterial cell membranes. TP also increased the permeability of the outer and inner membranes of P. aeruginosa and disrupted the cell membrane with the release of small cellular molecules. A proteomics approach based on two-dimensional gel electrophoresis and MALDI-TOF/TOF MS analysis was used to study the differences in the membrane proteins of TP-treated P. aeruginosa and those of control samples. Twenty-seven differentially expressed proteins were observed in the treated and the control groups. Most of the proteins identified by MALDI-TOF/TOF MS were enzymes (dihdrollpoamide dehydrogenase 50s ribosomal protein, and so on), which may have induced the metabolic disorder of the bacteria and resulted in their death.

Changes in growth performance, metabolic enzyme activities, and content of Fe, Cu, and Zn in liver and kidney of tilapia (Oreochromis niloticus) exposed to dietary Pb.

Tilapia (Oreochromis niloticus) were exposed to 0, 100, 400, and 800 microg/g concentrations of Pb in diet for 60 days, and changes in growth performance, metabolic enzyme activities, and essential trace elements (Fe, Cu, and Zn) content in liver and kidney were investigated. Daily weight gain, feed conversation ratio, and survival of tilapia were not significantly affected by dietary Pb. Alanine transaminase (ALT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) activities in liver and kidney were affected by dietary Pb in a dissimilar way: Pb concentration-related decreases in ALT, AST, and LDH activities were observed in kidney, while these enzyme activities in liver were stimulated in a Pb concentration-dependent manner. It was demonstrated that the inhibitory effects of dietary Pb on alkaline phosphatase, Na, K-adenosine triphosphatase (ATPase), Ca, and Mg-ATPase activities in both liver and kidney were Pb concentration-dependent. It was also indicated that the content of Fe, Cu, and Zn in liver and kidney decreased with the increasing dietary Pb concentrations. The results suggested that long-term dietary Pb exposure could affect metabolic enzyme activities and the content of Fe, Cu, and Zn in liver and kidney, whereas growth impairment was not observed in tilapia.