Three novel umami peptides derived from the alcohol extract of the Pacific oyster(Crassostrea gigas):identification,characterizations and interactions with T1R1/T1R3 taste receptors

Oyster(Crassostrea gigas),the main ingredient of oyster sauce,has a strong umami taste.In this study,three potential umami peptides,FLNQDEEAR(FR-9),FNKEE(FE-5),and EEFLK(EK-5),were identified and screened from the alcoholic extracts of the oyster using nano-HPLC-MS/MS analysis,i Umami-Scoring Card Method(i Umami-SCM)database and molecular docking(MD).Sensory evaluation and electronic tongue analysis were further used to confirm their tastes.The threshold of the three peptides ranged from 0.38 to 0.55 mg/m L.MD with umami receptors T1R1/T1R3 indicated that the electrostatic interaction and hydrogen bond interaction were the main forces involved.Besides,the Phe592 and Gln853 of T1R3 were the primary docking site for MD and played an important role in umami intensity.Peptides with two Glu residues at the terminus had stronger umami,especially at the C-terminus.These results contribute to the understanding of umami peptides in oysters and the interaction mechanism between umami peptides and umami receptors.

Non-Targeted Metabolomics Reveals the Metabolic Alterations in Response to Artificial Selective Breeding in the Fast-Growing Strains of Pacific Oyster

Pacific oyster(Crassostrea gigas)is one of the most important mollusks cultured all around the world.Selective breeding programs of Pacific oysters in China is initiated since 2006 and developed the genetically improved strain with fast-growing trait.However,little is known about the metabolic signatures of the fast-growing trait.In the present study,the non-targeted metabolomics was performed to analyze the metabolic signatures of adductor muscle tissue in one-year old Pacific oysters from fast-growing strain and the wild population.A total of 7767 and 10174 valid peaks were extracted and quantified in ESI^(+)and ESI^(−)modes,resulting in 399 and 381 annotated metabolites,respectively.PCA and OPLS-DA revealed that considerable separation among samples from fastgrowing strain and wild population,suggesting the differences in metabolic signatures.Meanwhile,81 significantly different metabolites(SDMs)were identified in the comparisons between fast-growing strain and wild population,based on the strict thresholds.It was found that there were highly correlation and conserved coordination among these SDMs.KEGG enrichment analysis indicated that the SDMs were tightly related to pantothenate and CoA biosynthesis,steroid hormone biosynthesis,riboflavin metabolism,and arginine and proline metabolism.Of them,the CoA biosynthesis and metabolism,affected by pantetheine and pantothenic acid,might be important for the growth of Pacific oysters under artificial selective breeding.The study provides the comprehensive views of metabolic signatures in response to artificially selective breeding,and is helpful to better understand the molecular mechanism of fastgrowing traits in Pacific oysters.

Characterisation of the Bacteria and Archaea Community Associated with Wild Oysters, At Three Possible Restoration Sites in the North Sea

With 85% of the global oyster reefs destroyed, there is an urgent need for large scale restoration to benefit from the ecosystem services provided by biogenic oyster reefs and their associated biodiversity, including microorganisms that drive marine biogeochemical cycles. This experiment established a baseline for the monitoring of the bacterial and archaeal community associated with wild oysters, using samples from their immediate environment of the Voordelta, with cohabiting Crassostrea gigas and Ostrea edulis, Duikplaats with only C. gigas attached to rocks, and the Dansk Skaldyrcentre, with no onsite oysters. The microbial profiling was carried out through DNA analysis of samples collected from the surfaces of oyster shells and their substrate, the sediment and seawater. Following 16S rRNA amplicon sequencing and bioinformatics, alpha indices implied high species abundance and diversity in sediment but low abundance in seawater. As expected, Proteobacteria, Bacteroidetes, Firmicutes and Thaumarchaeota dominated the top 20 OTUs. In the Voordelta, OTUs related to Colwellia, Shewanella and Psychrobium differentiated the oysters collected from a reef with those attached to rocks. Duikplaats were distinct for sulfur-oxidizers Sulfurimonas and sulfate-reducers from the Sva 0081 sediment group. Archaea were found mainly in sediments and the oyster associated microbiome, with greater abundance at the reef site, consisting mostly of Thaumarchaeota from the family Nitrosopumilaceae. The oyster free site displayed archaea in sediments only, and algal bloom indicator microorganisms from the Rhodobacteraceae, Flavobacteriaceae family and genus [Polaribacter] huanghezhanensis, in addition to the ascidian symbiotic partner, Synechococcus. This study suggests site specific microbiome shifts, influenced by the presence of oysters and the type of substrate.

Biological oyster shell waste enhances polyphenylene sulfide composites and endows them with antibacterial properties

To date,there is no research that deals with biological waste as fillers in polyphenylene sulfide(PPS).In this study,oyster shells were recycled and treated to prepare thermally-treated oyster shells(TOS),which were used as PPS fillers to make new bio-based antibacterial composite materials.The effect of varying the content of TOS was studied by means of structure and performance characterization.PPS/TOS composites were demonstrated to have an antibacterial effect on the growth of E coli and S.aureus.Qualitative analysis showed that when the TOS content was≥30%and 40%,the composite materials had an apparent inhibition zone.Quantitative analysis showed that the antibacterial activity increased with the TOS content.Fourier transform infrared spectroscopy indicated the formation of hydrogen bonds between the molecular chains of TOS and PPS and the occurrence of a coordination reaction.At 10%TOS,the composite tensile strength reached a maximum value of 72.5 MPa,which is 9.65%higher than that of pure PPS.The trend of bending properties is the same as that of tensile properties,showing that the maximum property was reached for the composite with 10%TOS.At the same time,the crystallinity and contact angle were the highest,and the permeability coefficient was the lowest.The fatigue test results indicated that for the composite with 10%TOS,the tensile strength was 23%lower than static tensile strength,and the yield strength was 10%lower than the static yield strength.The results of the study showed that TOS not only could reduce the cost of PPS,but also could impart antibacterial properties and enhance the mechanical and,barrier properties,the thermostability,as well as the crystallinity.

Near-infrared spectroscopy method for rapid proximate quantitative analysis of nutrient composition in Pacific oyster Crassostrea gigas

Glycogen,amino acids,fatty acids,and other nutrient components affect the flavor and nutritional quality of oysters.Methods based on near-infrared reflectance spectroscopy(NIRS)were developed to rapidly and proximately determine the nutrient content of the Pacific oyster Crassostreagigas.Samples of C.gigas from 19 costal sites were freeze-dried,ground,and scanned for spectral data collection using a Fourier transform NIR spectrometer(Thermo Fisher Scientific).NIRS models of glycogen and other nutrients were established using partial least squares,multiplication scattering correction first-order derivation,and Norris smoothing.The R_(C) values of the glycogen,fatty acids,amino acids,and taurine NIRS models were 0.9678,0.9312,0.9132,and 0.8928,respectively,and the residual prediction deviation(RPD)values of these components were 3.15,2.16,3.11,and 1.59,respectively,indicating a high correlation between the predicted and observed values,and that the models can be used in practice.The models were used to evaluate the nutrient compositions of 1278 oyster samples.Glycogen content was found to be positively correlated with fatty acids and negatively correlated with amino acids.The glycogen,amino acid,and taurine levels of C.gigas cultured in the subtidal and intertidal zones were also significantly different.This study suggests that C.gigas NIRS models can be a cost-effective alternative to traditional methods for the rapid and proximate analysis of various slaughter traits and may also contribute to future genetic and breeding-related studies in Pacific oysters.

Detection and quantification of Pb and Cr in oysters using laser-induced breakdown spectroscopy

The quantitative determination of heavy metals in aquatic products is of great importance for food security issues.Laser-induced breakdown spectroscopy(LIBS)has been used in a variety of foodstuff analysis,but is still limited by its low sensitivity when targeting trace heavy metals.In this work,we compare three sample enrichment methods,namely drying,carbonization,and ashing,for increasing detection sensitivity by LIBS analysis for Pb and Cr in oyster samples.The results demonstrate that carbonization can remove a significant amount of the contributions of organic elements C,H,N and O;meanwhile,the signals of the metallic elements such as Cu,Pb,Sr,Ca,Cr and Mg are enhanced by3–6 times after carbonization,and further enhanced by 5–9 times after ashing.Such enhancement is not only due to the more concentrated metallic elements in the sample compared to the dried ones,but also the unifying of the matter in carbonized and ashed samples from which higher plasma temperature and electron density are observed.This condition favors the detection of trace elements.According to the calibration curves with univariate and multivariate analysis,the ashing method is considered to be the best choice.The limits of detection of the ashing method are 0.52 mg kg-1 for Pb and0.08 mg kg-1 for Cr,which can detect the presence of heavy metals in the oysters exceeding the maximum limits of Pb and Cr required by the Chinese national standard.This method provides a promising application for the heavy metal contamination monitoring in the aquatic product industry.

The dual-function of bioactive peptides derived from oyster (Crassostrea gigas) proteins hydrolysates

Oysters(Crassostrea gigas)have a wide range of functionality due to their nutritional and bioactive components. However, the bioactive peptides of oyster proteins are rarely reported, particularly their antidiabetes effects and antioxidants. Oyster proteins were extracted from fresh oysters using phosphatebuffered saline and simulated gastrointestinal digestion was performed. The degree of hydrolysis(DH), structural characterization, molecular weight(Mw)distribution, free amino acid, anti-diabetic activity, and antioxidant activity were studied during in vitro simulated gastrointestinal digestion. The results showed that the α-glucosidase inhibitory activity, α-amylase inhibitory activity, DPPH radical scavenging activity, and ABTS radical scavenging activity of the oyster protein gastrointestinal digest were increased(P < 0.05)from 0 to 33.96%, from 9.17% to 44.22%, from 9.01 μg trolox/mg protein to 18.48 μg trolox/mg protein, and from 21.44 μg trolox/mg protein to 56.21 μg trolox/mg protein, respectively. Additionally, the DH, β-turn structure, fluorescence intensity, free amino acid, and short peptide content(Mw < 1 000 Da)increased in the simulated gastrointestinal digestion. These results indicate that the digestive hydrolysates obtained from oyster proteins could be used as natural anti-diabetic and antioxidant agents.

Preparation of Iron-Pillared Bentonite/Oyster Shell Composite and Phosphate Adsorption in Water

Iron-pillared bentonite(FB)was prepared by Fe(III)modified bentonite,and then the composites(FB-OS)were prepared by iron-pillared bentonite and oyster shell powder.The composites were characterized by FTIR,SEM,TGA,and EDS,and the phosphorus removal test was carried out.The results showed that FB-OS contained a large amount of CaO.Its structure was compact,but there were gaps in it.The maximum bending stress and compressive strength were 43.7 N and 0.927 MPa,respectively.The phosphorus removal test showed that the phosphorus removal rate of FB-OS was more than 90%,and measured the maximum adsorption capacity was 48.31 mg/g.A large amount of spherical products were produced on the surface and inside of FB-OS after phos-phorus removal,it was speculated that spherical products were amorphous calcium phosphate in the paper.Ana-lysis indicated that there was chemical adsorption during phosphorus removal.The kinetic equation of phosphorus adsorption by FB-OS was qt=10:193t/1+2:574t (R^(2)=0.995).The adsorption rate was mainly controlled by outerfilm diffusion and intraparticle diffusion.

Purification of β-Glucan of Oyster Mushroom (Pleurotus pulmonarius) and Its Application in Model Food

Mushrooms are recognized as sources of β-glucan. The aim of this study was to purify β-glucan from oyster mushrooms (Pleurotus pulmonarius) by alkaline extraction. The extracted β-glucan was added to a cracker recipe and the sensory, physical and estimated glycemic index of the final product were evaluated. Optimum extraction parameters were determined to be 80˚C, 90 minutes and 30% KOH. The β-glucan crackers had a glycemic index ranging from 88.07 to 67.32, while glucose had a glycemic index of 100. Based on the sensory study, it can be concluded that crackers enriched with β-glucan and possessing functional qualities may meet consumer expectations.

Heavy metals in oysters, mussels and clams collected from coastal sites along the Pearl River Delta,South China

Concentrations of 8 heavy metals: cadmium (Cd), copper (Cu), zinc (Zn), lead (Pb), nickel (Ni), chromium (Cr), antimony (Sb) and tin (Sn) were examined in 3 species of bivalves (Perna viridis, Crassostrea rivularis and Ruditapes philippinarum) collected from 25 sites along the Pearl River Delta coastal waters in the South China Sea from July to August 1996. In general, Cd, Cu, Zn and Sn concentrations in the three bivalve species collected from the Estuarine Zone were significantly higher than those collected from the Western and Eastern Zones of the Pearl River Delta, which are related to the existence of various anthropogenic activities in the catchment of the Pearl River Delta. The Western Estuarine Zone is mainly impacted by Cr, Ni and Cu contamination. In Victoria Harbor, heavy metal contamination is mainly due to Cu and Pb. Cd, Cu and Zn concentrations in oysters were significantly higher than those in mussels and clams. This could be explained by the fact that oysters live mainly in the Estuarine Zone of the Pearl River Delta which receives most of the polluting discharges from the catchment of the Delta. During turbid condition, heavy metals(soluble or adsorbed on suspended particulates) discharged from the Delta are filtered from the water column and subsequently accumulated into the soft body tissues of oysters. Heavy metal concentrations in the three bivalve species were compared with the maximum permissible levels of heavy metals in seafood regulated by the Public Health and Municipal Services Ordinance, Laws of Hong Kong, and it was revealed that Cd and Cr concentrations in the three bivalve species exceeded the upper limits. At certain hotspots in the Delta, the maximum acceptable daily load for Cd was also exceeded.

Comparative Study on the Antioxidant Activity of Peptides from Pearl Oyster(Pinctada martensii) Mantle Type V Collagen and Tilapia(Oreochromis niloticus) Scale Type Ⅰ Collagen

In this study, Pearl oyster mantle type V collagen(POMC) and tilapia scale type I collagen(TSC) were extracted and hydrolyzed by various proteases in order to obtain peptides. The antioxidant activity of the peptides was investigated by DPPH, hydroxyl radical scavenging experiments and a dynamic digestion model in vitro. The results show that there are significant differences in amino acid composition between POMC and TSC. The collagen peptides obtained from pearl oyster mantle(POMCP) by treating with alkaline protease exhibited higher antioxidant activity than that from tilapia scale(TSCP) treated with papaya protease, and both of them showed greater DPPH and hydroxyl radical scavenging activity than other peptides. After being separated via Sephadex G-25 chromatography, the M1 fraction isolated from POMCP, and the S1 fraction from TSCP with which both had higher molecular weights showed the strongest antioxidant activity than other fractions, and the M1 fraction exhibited stronger antioxidant activity than the S1 fraction in scavenging free-radicals and protecting cells from the oxidation damage. Furthermore, after treating the dynamic digestion system model in vitro, the DPPH and hydroxyl radical scavenging activity of the M1 fraction increased slightly. These results suggest that POMCP exhibits stronger antioxidant activity than TSCP, which means that PMOP may be a good candidate to be a potential natural antioxidant in the food-processing industry.

High Pressure Treatment Changes Spoilage Characteristics and Shelf Life of Pacific Oysters(Crassostrea gigas)During Refrigerated Storage

The effects of high pressure(HP) treatment on spoilage characteristic and shelf life extension of Pacific oysters(Crassostrea gigas) during refrigerated storage were studied.Results showed that HP treatment of 275 MPa for 3 min or 300 MPa for 2 min could achieve 100% full release of oyster adductor muscle,pressures higher than 350 MPa caused excessive release as the shells of oysters were broken,thus use of higher pressures should be cautious in oyster processing industry because of its adverse impact on the appearance of shells.HP treatment(300 MPa,2 min) was proper for the shucking of Pacific oyster(Crassostrea gigas) in China.This treatment caused no organoleptic disadvantage.Moreover,HP treatment resulted in obvious differences in biochemical spoilage indicators(p H,TVB-N and TBARS) changes and volatile compounds profile determined by electronic nose during storage.HP treatment(300 MPa,2 min) also led to a reduction of aerobic bacterial count(APC) by 1.27 log cycles.Furthermore,the APC values of oysters treated by HP were always lower than those of the control samples during storage.Based on the organoleptic,biochemical and microbiological indicators,shelf life of 6-8 d for control and 12 d for HP-treated oysters could be expected.HP treatment showed great potential in oyster processing and preservation.

Researches on the Treatment of Phosphorous Wastewater with Oyster Shells

Based on the analysis of adsorptive features of oyster shells,the researches on the treatment of phosphorous wastewater with oyster shells and the effect of temperature on phosphorus removal were carried out.XRD was used to characterize the crystalline phases,and the main component of oyster shells was shown to be CaCO3.When the pretreatment temperature reached 800 ℃,some CaCO3 decomposed into CaO.As the temperature was further raised,CaO increased gradually.Via SEM testing,the oyster shell was a kind of natural porous materials.The pore wall partially collapsed from 550 to 900 ℃.No obvious porous structure was found at 900 ℃.However,without preheating,the oyster shell phosphorous removal material can not adsorb the phosphorus.Pretreatment made calcium activate,thus greatly increasing the absorption of phosphorus.

Preparation, Characteristics, and Formation Mechanism of Oyster Peptide-Zinc Nanoparticles

Oyster peptide zinc nanoparticles (OPZNPs)(28 108 nm) were prepared in the presence of 0.5% 0.9% zinc sulfate at pH 6.0 11.0. The obtained nanoparticles exhibited uniform size distribution and spherical shapes. Nanoparticle characteristics, such as size, surface charge, and hydrophobicity, could be adjusted by controlling zinc sulfate concentration and environmental pH. In- creasing pH value or decreasing zinc sulfate concentration tended to reduce nanoparticle size and increase nanoparticle surface charge and hydrophobicity. OPZNPs presented good stability at near-neutral pH and could be stored for at least 20 days at 4℃. The results of the peptide conformation study and nanoparticle dissociation test proved that zinc ions and carboxyl groups are the key factors that affect OPZNP formation. The intermolecular combinations of carboxyl groups via zinc bridging facilitated the aggrega- tion of oyster peptides. Nanoparticle formation was accompanied by aggregate association and conformational changes. These changes included increments in β-sheets, especially intermolecular β-sheets, at the expense of α-helixes. Overall, this work provided a green alternative route for the synthesis of OPZNPs.

New Active Organic Substance in Oyster Shell Capable of Scavenging Oxygen Free Radicals with High Efficiency

A light purple organic active substance capable of scavenging hydroxyl radical（OH） with a high efficiency was extracted from Oyster shell at an extraction rate of 2.49%. It was found for the first time that this active substance may scavenge OH with the efficiency far higher than that of vitamin C. This active substance may scavenge also superoxide radical（O2） although the scavenging efficiency is far lower than that of vitamin C. Infrared spectrometry and routine chemical analysis primarily reveal that this active substance belongs to glycoprotein.

Mass Selection for Growth Improvement in Black Shell Line of Pacific Oyster Crassostrea gigas

The Pacific oyster,Crassostrea gigas,naturally distributing along the coast of northwest Pacific,is one of the most important bivalve species due to its high economic value and fecundity.In China,we have initiated a selective breeding program on both shell color and growth rate of C.gigas since 2010.A black shell line was obtained through four-generation family selection.In this study,mass selection for growth improvement was conducted in the sixth generation and seventh generation of black shell lines.To assess the progress of potential genetic improvement,the progeny of two generations of black shell lines were selected to evaluate their shell heights via a 450-day farming experiment.As the results,after growing for 450 days,the sixth generation and seventh generation of selected lines were 9.03% and 11.42% larger than the control lines,respectively.During the grow-out stage,the genetic gain of two generations was 8.82%±0.18% and 11.54%±0.43%,respectively;and the corresponding realized heritability was 0.45±0.04 and 0.41±0.04,respectively.These results indicated that the mass selection for shell height achieved steady progress in the two generations of C.gigas.

论Oysters

林奈于1758年建立了Ostrea (s.l.)属。此后的两百多年中经拉马克(Lamark)、杜威尔(Douville)、乌雅洛夫(Vyalov)、斯坦兹尔(Stenzel)等许多学者的深入研究,不断修正、补充与完善,原来的Oatrea (s.l.)属现已扩展为牡蛎超科(SuperfamilyOstreacea),人们通常称之为Oysters(牡蛎类)。目前对Oysters的特征、分类、生态、起源、演化等方面的认识正不断深化,但仍存在不少分歧。

Researches on the Removal of Pb(Ⅱ) by Hydrothermally Modified Oyster Shells

A novel wastewater purification material was prepared by a hydrothermal method. It was mainly made from oyster shells with the merits of long service time, large surface area, high lead removal efficiency and excellent recyclable properties. The technological conditions were decided respectively based on the lead removal efficiencies. At pH = 5 and 30 ℃, for the wastewater with the initial concentration of Pb2＋ to be 5 mg/L, adsorption time 24 h, and 1 mg to 40 mL of mass ratio between adsorbent and Pb2＋, the maximum adsorption capacity can reach 0.19 mg/g. The lead removal material prepared by hydrothermal method has excellent recycle performance. The equilibrium adsorption capacity can get to 9.71 mg/g and the average Pb2＋ removal rate is as high as 66.39%. After reusing for 60 times, the SEM observation shows that the hydrates of reticular formation is formed after hydrothermal modification, which provides a good attachment position for Pb2＋, indicating the physical adsorption is dominant.

Structure Characterization and Dephosphorization Effect Analysis of Oyster Shell-silica Micropowder Waste Water Dephosphorization Materials

In this work,the effects of pH value of waste water and initial concentration of phosphorus on dephosphorization materials were investigated.The materials were prepared by shaping,sintering and hydrothermal reshaping oyster shell and silica micro-powder.Different concentrations of phosphorus-contained waste water were simulated with potassium dihydrogen phosphate solution,the effect of dephosphorization was tested with phosphomolybdenum blue spectrophotometer method,and the crystal phase and microstructure of materials were characterized by XRD and SEM methods. It was indicated that dephosphorization was completed in 6 h when the initial phosphorus concentration in waste water was lower than 15 mg/L, and the dephosphorization time prolonged as the increase of phosphorus concentration. It was observed that the pH value of waste water influenced dephosphorization significantly, and neutral subalkalic environment favored dephosphorization. When the pH value was 11, the efficiency of dephosphozation was the greatest. For waste water with an initial concentration of 20 mg/L, the dephosphozation rate is close to 100% in8 h.

Study on the Structures and Properties of Copper Removal Adsorbent Prepared from Sinter-free Pulverized Oyster Shell Materials

A hollow tubular copper removal adsorbent was prepared with oyster shell and cement as the main raw materials. The effects of different formulas, different initial copper concentrations and different pH values of samples on the copper removal efficiency were investigated to determine the optimal conditions for copper removal. The content of copper in the wastewater is determined by Atomic Absorption Spectrophotometer. The microstructure and elemental composition of the samples were characterized by scanning electron microscopy（SEM） and EDS. As a result, the formula with the content of cement to be 8 wt% and the oyster shell powder of 92 wt% is optimal. Under the condition of 30 ℃, when the pH value was 9.0, the Cu2＋ adsorption capacity of the sample could reach 0.59 mg/g at 48 h. SEM analysis revealed that there are abundant pores in the sample, which is beneficial for Cu2＋ absorption on the adsorbent.