The detection and regulation advances of food allergens

Food allergy has become an important food quality and safety issue,posing a challenge to the food industry and affecting consumer health.On the one hand,from the perspective of food processing industry,the diversity of food raw material ingredients,exogenous additives,and processing forms make the presence of allergens in modern food processing more complex.In addition,due to the lack of allergen identification,effective detection and allergenicity evaluation systems,there are serious deficiencies in the current theories and techniques for food allergen screening and detection,tracking and prediction,intervention and control;On the other hand,from the perspective of public health,meeting consumers'right to know whether there are raw materials containing food allergens in processed foods,and improving the credibility of government and people's satisfaction have become urgent matters;In addition,as people come into contact with more and more new borne novel foods,the probability of food allergy is also increasing.The food safety and health problems induced by increasingly complex,widespread and severe food allergy are difficult to avoid.In view of this,in response to the increasingly serious food allergy issues,this paper introduced the detection methods of food allergens,summarized the reduction and control techniques of food allergens,and elaborated hypoallergenic foods,which aims to provide the basis for preventing and controlling food allergy and ensuring the physical health of food allergy patients.

Expression and Characterization of a Novelλ-Carrageenase Cgl150A_Wa from Wenyingzhuangia aestuarii

λ-Carrageenan is a highly sulfated polysaccharide alternating of 1,4-O-α-D-galactopyranose-2,6-sulfate(D2S,6S)and 1,3-O-β-D-galactopyranose-2-sulfate(G2S).λ-Carrageenases are desirable tools forλ-carrageenan degradation.Based on the genome mining,a novelλ-carrageenase Cgl150A_Wa was cloned from the bacterium Wenyingzhuangia aestuarii and expressed in Escherichia coli.Cgl150A_Wa was an endo-acting enzyme and exhibited its maximum activity at 30℃and pH 8.0.By employing a glycomics strategy that combined ultra-performance liquid chromatography-mass spectrometry analysis and glycoinformatics,Cgl150A_Wa was proven to degradeλ-carrageenan octaose and hexaose,and the major hydrolysis product of Cgl150A_Wa wasλ-carrageenan tetrose.In addition to the typicalλ-carrageenan motifs,the active center of Cgl150A_Wa might tolerate desulfatedλ-carrageenan motifs.Cgl150A_Wa is a potential biotechnological tool for preparingλ-carrageenan oligosaccharides and structural investigation.

Effects of alcohol on digestion,absorption and metabolism of sea cucumber saponins in healthy mice

Sea cucumber saponins have attracted more attention in recent years due to biological activities.It is a popular practice to soak sea cucumber in Baijiu at home and being applied to industrial manufacturing in China.However,knowledge of the effect of alcohol on the absorption and metabolism of sea cucumber saponins is limited.The effects of alcohol on digestion,absorption and metabolism of sea cucumber saponins in BALB/c mice were investigated after gavage and tail intravenous injection.The results showed that the content of saponins in serum and liver was significantly higher under the influence of alcohol than that in the control group after oral administration.Alcohol promoted the absorption of sea cucumber saponins prototype as well as inhibited the process of saponins being transformed into deglycositic metabolites in the small intestine.Moreover,sea cucumber saponins remained in circulation for a long time and alcohol slowed down the clearance of sea cucumber saponins under the influence of alcohol after intravenous injection.This confirmed the feasibility of marinating sea cucumber in Baijiu to improve the efficacy of saponins and provides an important theoretical basis for the utilization of sea cucumber and the development of sea cucumber liquor.

Trimethylamine N-oxide generation process was influenced by the proportion and source of macronutrients in the diet

Trimethylamine N-oxide(TMAO)is a risk factor of various chronic diseases,which was produced by metabolism from precursors to trimethylamine(TMA)in gut and the oxidation from TMA in liver.The TMA generation was influenced by diet,mainly due to the rich TMAO precursors in diet.However,it was still unclear that the effects of different proportion and source of macronutrients in different dietary pattern on the production process of TMAO.Here,the generation of TMA from precursors and TMAO from TMA was determined after single oral choline chloride and intraperitoneal injection TMA,respectively,in mice fed with carbohydrates,proteins and fats in different proportion and sources.The results suggested that the generation of TMAO was increased by low non-meat protein and high fat via enhancing the production of TMAO from TMA,and decreased by plant protein and refined sugar via reducing TMA production from precursors in gut and TMAO transformation from TMA in liver.The high fat and high sugar diets accelerating the development of atherosclerosis did not increase the production of TMAO,the risk factor for atherosclerosis,which indicated that the dietary compositions rather than the elevated TMAO level might be a more key risk factor for atherosclerosis.

Analysis of quality-related proteins in golden pompano(Trachinotus ovatus)fillets with modified atmosphere packaging under superchilling storage

Here,we aimed to study the changes in proteome of golden pompano fillets during post-mortem storage.Tandem mass tags(TMT)-labeled quantitative proteomic strategy was applied to investigate the relationships between protein changes and quality characteristics of modified atmosphere packaging(MAP)fillets during superchilling(-3°C)storage.Scanning electron microscopy was used to show that the muscle histology microstructure of fillets was damaged to varying degrees,and low-field nuclear magnetic resonance was used to find that the immobilized water and free water in the muscle of fillets changed significantly.Total sulfhydryl content,TCA-soluble peptides and Ca2+-ATPase activity also showed that the fillet protein had a deterioration by oxidation and denaturation.The Fresh(FS),MAP,and air packaging(AP)groups were set.Total of 150 proteins were identified as differential abundant proteins(DAPs)in MAP/FS,while 209 DAPs were in AP/FS group.The KEGG pathway analysis indicated that most DAPs were involved in binding proteins and protein turnover.Correlation analysis found that 52 DAPs were correlated with quality traits.Among them,8 highly correlated DAPs are expected to be used as potential quality markers for protein oxidation and water-holding capacity.These results provide a further understanding of the muscle deterioration mechanism of packaging golden pompano fillets during superchilling.

Biotransformation of Shrimp Wastes by Bacillus subtilis OKF04 and Evaluation of Growth Promoting Effect in Crop Planting

In this study,we proposed a reliable and sustainable technique for the clean utilization of shrimp wastes,which can yield a solid inoculant of Bacillus subtilis OKF04 containing micronutrients at low cost without the risk of contamination.Study of the culture conditions revealed that the head of shrimp Litopenaus vannamei and the wheat bran acted as suitable substrates for the growth of B.subtilis OKF04.With 60%initial moisture content,30℃culture temperature,and 5%inoculation amount,followed by 48 hours of fermentation and 0.5%soluble starch added during the drying process(50℃for 6h),a solid B.subtilis OKF04 inoculant with a spore amount of 2.4×10^(10)CFU g^(-1)and a high amino acid content was obtained.The solid B.subtilis OKF04 inoculant was applied to cultivate pakchoi under pot experiment.As the result,of adding to,the size of stems and leaves,nutritional composition,and physiological activity of pakchoi were significantly(P<0.05)enhanced by solid B.subtilis OKF04 inoculant.B.subtilis OKF04 also significantly(P<0.05)increased the soil’s nutrient content and improved its microbial composition.Furthermore,pakchoi cultivated with a low dose of solid B.subtilis OKF04 inoculant(0.05 g kg^(-1)soil)resulted in the best results.This study provides a new method for the preparation of microbial inoculants with solid waste shrimp heads.

Extraction of Natural Nanostructured Hydroxyapatite from Pacific Cod(Gadus macrocephalus)Bone with a Thermostable Collagenolytic Protease and Its ex vivo Intestinal Bioavailability

Natural nano-hydroxyapatite(HA)was extracted from Pacific cod(Gadus macrocephalus)bone with a thermostable col-lagenolytic protease in the present study.Conditions for the enzymatic reaction were optimized to be 60℃and pH 7.0,and a desir-able extraction efficiency was achieved by using the crude collagenolytic protease.Dynamic light scattering,transmission electron microscopy and energy-dispersive X-ray analysis revealed that nano-HA are anionic spherical(about 110nm)particles mainly com-prised of calcium and phosphorus at an approximate ratio of 5:3.As evaluated with the mouse ex vivo intestinal segments,the extracted nano-HA displayed comparable level of intestinal bioavailability to the positive control CaCl_(2).By treating with inhibitors(NaN3,ami-loride)and low temperature(4℃),clathrin-mediated endocytosis was assumed to involve the intestinal absorption of nano-HA.Over-all,the application of thermostable collagenolytic protease is proved to be a promising alternative method for nano-HA extraction from natural resource with improved ecological and biological value.

Cloning,Expressing and Characterizing a Novel Cold-Adapted Laminarinase from Marine Bacterium Wenyingzhuangia aestuarii OF219

Laminarinases reveal potential application in the field of food and biotechnology.In this research,a novel GH16 family laminarinase,designated as Lam16A_Wa,was cloned from the genome of marine bacterium Wenyingzhuangia aestuarii OF219 and expressed in Escherichia coli.Lam16A_Wa demonstrates a relatively low optimal reaction temperature(35℃)and a cold-adapted feature.Its optimal pH value is 6.0 and is stable in a broad pH range from 3.0 to 11.0.A glycomics strategy was employed to investigate the hydrolytic pattern of Lam16A_Wa.The enzyme was confirmed as a random endo-acting glycoside hydrolase.Its minimum substrate was laminarin pentasaccharide,and the major final products are oligosaccharides,including disaccharide to pentasaccharide.The Lam16A_Wa provides a novel and well-defined tool for the molecular tailoring of laminarin.

Echinoside A from Pearsonothuria graeffei Exert the Cytotoxicity to MDA-MB-231 Cells via Mitochondrial Membrane and Modulation of PI3K/Akt/mTOR Pathway

A kind of triterpene glycosides echinoside A(EA)was extracted from sea cucumber Pearsonothuria graeffei,and its yield was about 0.78%.The purity of EA was 99.0%,and its molecular weight was 1206 Da.EA was a linear tetrasaccharide attached to a pentacyclic triterpene aglycon.It inhibited the growth of MDA-MB-231 cells in vitro.The antitumor effect was related to elevate ROS level,decrease mitochondrial membrane potential,enhance caspase-3 expression,induce cells apoptosis and arrest cell cycle at G2/M phase.EA also dose-dependently suppressed the expressions of phophorylation proteins p-PI3K,p-Akt,and p-mTOR as analyzed by western blotting.These results suggested that EA caused MDA-MB-231 cells apoptosis via intrinsic mitochondrial and PI3K/Akt/mTOR pathway.EA can be a potential anti-breast cancer agent to enhance the clinical efficacy.

Novel insight into the formation and inhibition mechanism of dipeptidyl peptidase-Ⅳ inhibitory peptides from fermented mandarin fish(Chouguiyu)

Fermented foods are a potential source to produce novel dipeptidyl peptidase-IV inhibitory peptides(D4IPs).In this study,the fermented mandarin fish(Chouguiyu)was used to screen D4IPs and their formation mechanism was studied by metagenomics and peptidomics.A total of 400 D4IPs with DPP-IV inhibition structure and high hydrophobicity were identified.The correlation network map showed that Lactococcus,Bacillus,Lysobacter,Pelagivirga,Kocuria,Escherichia,Streptococcus,and Peptostreptococcus were significantly correlated with the most D4IPs.Four stable D4IPs,including KAGARALTDAETAT,GEKVDFDDIQK,VVDADEMYLKGK,and GQKDSYVGDEAQ were respectively from the precursor proteins parvalbumin,troponin,myosin,and actin,and were mainly formed by the hydrolysis of subtilisin(EC 3.4.21.62),aspartic proteinase(EC 3.4.23.1),thermolysin(EC 3.4.24.27),oligopeptidase B(EC 3.4.21.83),and proteinase P1(EC 3.4.21.96)from Bacillus,Kocuria,Lysobacter,Lactococcus,and Peptostreptococcus.The inhibition mainly resulted from the hydrogen bond and salt bridge between D4IPs and DPP-IV enzyme.This study provides important information on the proteases and related microbial strains to directionally prepare D4IPs in Chouguiyu.

Advanced Glycation Endproducts in 35 Types of Seafood Products Consumed in Eastern China

Advanced glycation endproducts(AGEs) have been recognized as hazards in processed foods that can induce chronic diseases such as cardiovascular disease, diabetes, and diabetic nephropathy. In this study, we investigated the AGEs contents of 35 types of industrial seafood products that are consumed frequently in eastern China. Total fluorescent AGEs level and Nε-carboxymethyl-lysine(CML) content were evaluated by fluorescence spectrophotometry and gas chromatography-mass spectrometry(GC-MS), respectively. The level of total fluorescent AGEs in seafood samples ranged from 39.37 to 1178.3 AU, and was higher in canned and packaged instant aquatic products that were processed at high temperatures. The CML content in seafood samples ranged from 44.8 to 439.1 mg per kg dried sample, and was higher in roasted seafood samples. The total fluorescent AGEs and CML content increased when seafood underwent high-temperature processing, but did not show an obvious correlation. The present study suggested that commonly consumed seafood contains different levels of AGEs, and the seafood processed at high temperatures always displays a high level of either AGEs or CML.

Purification and Characterization of an Alkaline Protease from Micrococcus sp.Isolated from the South China Sea

Protease is wildly used in various fields,such as food,medicine,washing,leather,cosmetics and other industrial fields.In this study,an alkaline protease secreted by Micrococcus NH54PC02 isolated from the South China Sea was purified and characterized.The growth curve and enzyme activity curve indicated that the cell reached a maximum concentration at the 30 th hour and the enzyme activity reached the maximum value at the 36 th hour.The protease was purified with 3 steps involving ammonium sulfate precipitation,ion-exchange chromatography and hydrophobic chromatography with 8.22-fold increase in specific activity and 23.68% increase in the recovery.The molecular mass of the protease was estimated to be 25 k Da by SDS-PAGE analysis.The optimum temperature and p H for the protease activity were 50℃ and pH 10.0,respectively.The protease showed a strong stability in a wide range of pH values ranging from 6.0–11.0,and maintained 90% enzyme activity in strong alkaline environment with p H 11.0.Inhibitor trials indicated that the protease might be serine protease.But it also possessed the characteristic of metalloprotease as it could be strongly inhibited by EDTA and strongly stimulated by Mn^(2+).Evaluation of matrix-assisted laser desorption ionization/time-of-flight MS(MALDI-TOF-TOF/MS) showed that the protease might belong to the peptidase S8 family.

Adsorption of Heavy Metal Ions,Dyes and Proteins by Chitosan Composites and Derivatives-A Review

Chitosan composites and derivatives have gained wide attentions as effective biosorbents due to their low costs and high contents of amino and hydroxyl functional groups.They have showed significant potentials of removing metal ions,dyes and proteins from various media.Chemical modifications that lead to the formation of the chitosan derivatives and chitosan composites have been extensively studied and widely reported in literatures.The aims of this review were to summarize the important information of the bioactivities of chitosan,highlight the various preparation methods of chitosan-based active biosorbents,and outline its potential applications in the adsorption of heavy metal ions,dyes and proteins from wastewater and aqueous solutions.

Protective Effect of Gelatin and Gelatin Hydrolysate from Salmon Skin on UV Irradiation-Induced Photoaging of Mice Skin

The objective of this study was to investigate the effect of gelatin(SG) isolated from salmon skin and its hydrolysate(SGH) on photoaging skin, and the mechanism responsible for anti-photoaging. The average molecular weights of SG and SGH were 65 k Da and 873 Da, respectively. The amino acid compositions of SG and SGH were similar. Both of them were abundant in hydrophobic amino acids. Twenty-five peptides were identified from SGH. SG and SGH could improve UV irradiation-induced pathological changes of macroscopical tissue texture and skin morphology. Hydroxyproline content is an indicator of matrix collagen content, SG and SGH could inhibit the decrease of hydroxyproline content in photoaging skin in a dose dependent manner. In addition, SG and SGH could alleviate UV irradiation-induced oxidative damages to skin by increasing the activities of total superoxide dismutase(T-SOD), glutathione peroxidase(GSH-Px) and catalase(CAT), increasing the content of glutathione(GSH) and decreasing the content of malonaldehyde(MDA). Moreover, SG and SGH could enhance immune regulation system by increasing the thymus index. Thus, the anti-photoaging mechanisms of SG and SGH were by inhibiting the depletion of antioxidant defense components, involving in the synthesis of collagen and enhancing the function of immune system. Besides, SGH showed a better result in protecting skin from photoaging than SG.

Changes in the Contents of ATP and Its Related Breakdown Compounds in Various Tissues of Oyster During Frozen Storage

The changes in the contents of adenosine 5’-triphosphate (ATP) and its related breakdown compounds were investigated in the adductor muscle, mantle, gill and body trunk of oyster (Crassostrea gigas) during frozen storage at -20℃ and -30℃ and compared with that of the fresh oyster. The investigation was performed using an HPLC system. Different extents of ATP decomposi- tion were found in various tissues frozen at the two temperatures. The K, K’ and A.E.C values were calculated as the chemical fresh- ness indices. Considering the results of sensory evaluation, the A.E.C. value in body trunk at -20℃ and -30℃ could be used as the best freshness index for frozen oyster.

Rheological and Structural Properties of Sea Cucumber Stichopus japonicus During Heat Treatment

Changes in tissue structure, rheological properties and water content of raw and heated sea cucumber meat werestudied. Sea cucumber Stichopus japonicus was heated at 25°C, 70°C and 100 °C water for 5 min. The structural changeswere observed using a light microscope and the rheological parameters (rupture strength, adhesive strength and deformation)determined using a texture meter. Microscopic photograph revealed that the structural change of heated meat was greaterthan that of raw meat. The rupture strength, adhesive strength and deformation of raw meat were smaller than those of theheated meat. Meanwhile, rheological parameters showed positive correlation with heating temperature. These changes aremainly caused by thermal denaturation and gelatinization of collagen during heating. These changes were also evidenced inobservations using a light microscope and differential scanning calorimetry.

Assessment and Comparison of in vitro Immunoregulatory Activity of Three Astaxanthin Stereoisomers

In recent years,the immune-modulatory role of all-trans astaxanthin from different pigment sources has been studied.It was reported that all-trans astaxanthin might exist as three stereoisomers,and the composition of all-trans stereoisomers in natural materials differs from that of synthetic products.However,the different biological effects of various all-trans stereoisomers still remain unclear.In the present study,we evaluated the bioactivity of three astaxanthin stereoisomers,(3S,3'S)-trans-,(3R,3'R)-transand meso-trans-astaxanthin,in regulating cell-mediated immune response using mice lymphocytes and peritoneal exudates cells(PECs) systems.After the treatment with three astaxanthin stereoisomers(20 μmol L-1),the lymphocyte proliferation capacity,neutral red phagocytosis of PECs and natural killer(NK) cell cytotoxic activity were comparatively assessed.The results showed that all three astaxanthin stereoisomers significantly promoted lymphocyte proliferation,phagocytic capacity of PECs,and cytotoxic activity of NK cells.Moreover,the(3S,3'S)-trans-astaxanthin exhibited a much higher response than others.

Isolation and Characterization of Collagen from Squid(Ommastrephes bartrami) Skin

Collagen of squid(Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen(ASC) and pepsin-solubilized collagen(PSC) were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type Ⅰ collagen,containing α1 and α2 chains. FTIR(fourier transform infrared spectroscopy) investigations confirmed the existence of helical arrangements in PSC of squid skin. The denaturation temperature(Td) and shrinkage temperature(Ts) of PSC were 29.4℃ and 52.8℃,respectively.

Degradation of Extraction from Seaweed and Its Complex with Rare Earths for Organophosphorous Pesticides

On the basis of several experiments carried out in China, it was proved that both seaweed liquid fertilizer and rare earth （RE） could promote the growth of crops and increase their yield. The effects of extraction from seaweed and its complex with RE on the degradation of organophosphorous pesticides and the yield of vegetables were investigated. The resuits showed that the extract and its complex with RE could degrade organophosphorous pesticides in neutral solvent. The residues of the pesticides treated by the extract decreased by 96.88 %, 52.30%, 49.52%, and 22.88 %, respectively, for chlorpyrifos, dichlorvos, omethoate, and dimethoate, and those by the complex decreased by 95.99%, 54.23%, 48.79%, and 25.66%, respectively, when compared with the control. The residues of chlorpyrifos and dimethoate in spinach sprayed with the complex were decreased by 90.64% and 76.56%, respectively, compared with those in spinach from control plots when the interval between spraying and sampling was 8 d. The fresh weight of brassica chinensis and cabbage increased by 28.62% and 18.72%, and their dry weight increased by 44.49% and 14.74%, respectively, compared with those of the controls. The chlorpyrifos and dimethoate residues in brasscia chinese were decreased by 36.36% and 50.00%, respectively, and their rate of decrease in cabbage was 40.00% and 75.00%, respectively, on 5th day after spraying with the complex, when compared with those in the vegetable from control. These results suggest that this complex can increase the agricultural productivity and reduce the use of pesticide residues in the production of vegetables.

Identification and Characterization of a New IgE-binding Protein in Mackerel (Scomber japonicus) by MALDI-TOF-MS

As fish is one source of the ‘big eight’ food allergens,the prevalence of fish allergy has increased over the past few years.In order to better understand fish allergy,it is necessary to identify fish allergens.Based on the sera from fish-allergenic patients,a 28 kDa protein from local mackerel (Scomber japonicus),which has not been reported as a fish allergen,was found to be reactive with most of the patients’ sera.The 28 kDa protein was analyzed by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry).Mascot search in NCBI database (Date: 08/07/2010) showed that the top protein matched,i.e.triosephosphate isomerase (TPI) from Xiphophorus maculatus and Poecilia reticulata,had a mowse (molecular weight search) score of 98.In addition,TPI from Epinephelus coioides also matched this mackerel protein with a mowse score of 96.Because TPI is con-sidered as an allergen in other non-fish organisms,such as lychee,wheat,latex,archaeopotamobius (Archaeopotamobius sibiriensis) and crangon (Crangon crangon),we consider that it may also be an allergen in mackerel.