Production of the Blood Pressure Lowing Peptides from Brown Alga （ Undaria pinnatifida ）

Brown alga （ Undaria pinnatifida） was treated with alginate lyase and hydrolyzed using 17 kinds of proteases and the inhibitory activity of the hydrolysates for the angiotensin-I-converting enzyme （ACE） was measured. Four hydrolysates with potent ACE-inhibitory activity were administered singly and orally to spontaneously hypertensive rats （SHRs）. The systolic blood pressure of SHRs decreases significantly after single oral administration of the brown alga hydrolysates by pro- tease S ＇ Amano＇ （from Bacillus stearothermophilus） at the concentration of 10 （mg protein） （kg body weight）^ - 1. In the 17 weeks of feeding experiment, 7-week-old SHRs were fed standard diet supplemented with the brown alga hydrolysates for 10 weeks. In SHRs fed 1.0 and 0.1% brown alga hydrolysates, elevating of systolic bloodpressure was significantly suppressed for 7 weeks. To elucidate the active components, the brown alga hydrolysates were fractionated by 1-butanol extraction and HPLC on a reverse-phase column. Seven kinds of ACE-inhibitory peptides were isolated and identified by amino acid composition analysis, sequence analysis, and LC-MS with the results Val-Tyr, Ile-Tyr, Ala-Trp, Phe-Tyr, Val-Trp, Ile-Trp, and Leu-Trp. Each peptide was determined to have an antihypertensive effect after a single oral administration in SHRs. The brown alga hydrolysates were also confirmed to decrease the blood pressure in humans.     

罗非鱼鱼鳞胶制备降血压胶原肽的工艺及酶解物活性

以罗非鱼鱼鳞胶为原料,制备对血管紧张素转换酶(ACE)有抑制活性的降血压肽。以水解度(DH)及ACE抑制率为指标,筛选水解鱼鳞胶的最适酶种,并对其水解工艺条件进行优化;采用超滤技术对所制备的降血压肽进行分离纯化,比较不同组分的ACE抑制率,测定高ACE抑制率组分的氨基酸组成,并运用MALDI-TOF-MS法分析其分子质量分布。结果表明,复合蛋白酶是水解鱼鳞胶最适酶种,在50℃、pH7.5、料液质量体积比1∶5、酶与底物质量比(mE/mS)1.5%条件下,酶解6 h,酶解液的ACE抑制作用最强,其水解度DH 73%,ACE的抑制率76.15%。酶解液经超滤分离所获得的分子质量小于2 000 u的组分对ACE抑制率最高,达83.15%;该组分的氨基酸主要含有Gly和Pro,含量分别为22.04%和11.40%;该组分分子质量范围为1 300~2 000 u。     

Amino acid composition and functional properties of giant red sea cucumber (Parastichopus californicus) collagen hydrolysates

Giant red sea cucumber (Parastichopus californicus) is an under-utilized species due to its high tendency to autolysis.The aim of this study was to evaluate the functional properties of collagen hydrolysates from this species.The degree of hydrolysis (DH),amino acid composition,SDS-PAGE,emulsion activity index (EAI),emulsion stability index (ESI),foam expansion (FE),and foam stability (FS) of hydrolysates were investigated.The effects of pH on the EAI,ESI FE and FS of hydrolysates were also inves-tigated.The results indicated that the β and α1 chains of the collagen were effectively hydrolyzed by trypsin at 50℃ with an En-zyme/Substrate (E/S) ration of 1:20 (w:w).The DH of collagen was up to 17.3% after 3 h hydrolysis with trypsin.The hydrolysates had a molecular weight distribution of 1.1 17 kDa,and were abundant in glycine (Gly),proline (Pro),glutamic acid (Glu),alanine (Ala) and hydroxyproline (Hyp) residues.The hydrolysates were fractionated into three fractions (< 3 kDa,3 10 kDa,and > 10 kDa),and the fraction of 3 10 kDa exhibited a higher EAI value than the fraction of > 10 kDa (P<0.05).The fraction of > 10 kDa had higher FE and FS values than other fractions (P<0.05).The pH had an important effect on the EAI,ESI,FE and FS.All the fractions showed undesirable emulsion and forming properties at pH 4.0.Under pH 7.0 and pH 10.0,the 3 10 kDa fraction showed higher EAI value and the fraction of > 10 kDa showed higher FE value,respectively.They are hoped to be utilized as functional ingredients in food and nutraceutical industries.     

Production of the Blood Pressure Lowing Peptides from Brown Alga ( Undaria pinnatifida )

1 IntroductionIncreased blood pressure appears to be one of theprimary risk factors of circulatory organ diseases suchas encepharo-apoplexy, encepharo-infarction and cardi-ac infraction. Angiotensin-I-converting enzyme (ACE)plays an important role in the rennin-angiotensin sys-tem by regulating blood pressure. Antihypertensivedrugs such as captopril and enalapril are potent ACEinhibitors (Ondetti et al., 1977). Recently, severalinhibitory peptides derived from food proteins havebeen isolat…     

Study on application of polyenzyme method to offal of Harengula zunasi

The new polyenzyme method for making gravy fromHarengula zunasi offal involves protein enzymolysis with flavorase after proper alkaline and neutral protease levels were established by orthogonal trials to select the best hydrolytic conditions for processing offal with alkaline and neutral protease. The conditions for the polyenzyme method were pH of 7.0, temperature of 50°C, alkaline and neutral protease concentration of 1.5% respectively, hydrolysis time of 120 min, and flavorase concentration of 2.0%, for 60 min. The new gravy-making technology yields a nutritious and delicious gravy containing 40.3% of total essential amino acids, with delicious amino acids Glu, Asp, Gly, Ala, Pro and Ser comprising 49.5%, total and amino nitrogen being respectively 1.9 and 1.1 g/100 g (amino acid nitrogen being 61.0% of total nitrogen), The polyenzyme method was used to make 14.8% protein gravy fromHarengula zunasi offal. In addition, inorganic elements, the phosphorus content is the highest. This project was aided financially by the Guangdong Science and Technology Office (No. 2KM06002S).     

Optimization of dilute acid hydrolysis of Enteromorpha

Acid hydrolysis is a simple and direct way to hydrolyze polysaccharides in biomass into fermentable sugars. To produce fermentable sugars effectively and economically for fuel ethanol, we have investigated the hydrolysis of Enteromorpha using acids that are typically used to hydrolyze biomass:H2SO4, HCl, H3PO4 and C4H4O4 (maleic acid). 5%(w/w) Enteromorpha biomass was treated for different times (30, 60, and 90 min) and with different acid concentrations (0.6, 1.0, 1.4, 1.8, and 2.2%, w/w) at 121°C. H2SO4 was the most effective acid in this experiment. We then analyzed the hydrolysis process in H2SO4 in detail using high performance liquid chromatography. At a sulfuric acid concentration of 1.8% and treatment time of 60 min, the yield of ethanol fermentable sugars (glucose and xylose) was high, (230.5 mg/g dry biomass, comprising 175.2 mg/g glucose and 55.3 mg/g xylose), with 48.6% of total reducing sugars being ethanol fermentable. Therefore, Enteromorpha could be a good candidate for production of fuel ethanol. In future work, the effects of temperature and biomass concentration on hydrolysis, and also the fermentation of the hydrolysates to ethanol fuel should be focused on.     

Study on Application of Polyenzyme Method to Offal of Harengula zunasi

The new polyenzyme method for making gravy from Harengula zunasi offal involves protein enzymolysis with flavorase after proper alkaline and neutral protease levels were established by orthogonal trials to select the best hydrolytic conditions for processing offal with alkaline and neutral protease. The conditions for the polyenzyme method were pH of 7.0, temperature of 50℃ , alkaline and neutral protease concentration of 1.5% respectively, hydrolysis time of 120 min, andflavorase concentration of 2.0% , for 60 min. The new gravy-making technology yields a nutritious and delicious gravy containing 40.3 % of total essential amino acids, with delicious amino acids Glu, Asp, Gly, Ala, Pro and Ser comprising 49.5% , total and amino nitrogen being respectively1.9 and 1.1 g/100 g (amino acid nitrogen being 61.0% of total nitrogen), The polyenzyme method was used to make 14.8% protein gravy from Harengula zunasi offal. In addition, in inorganic elements, the phosphorus content is the highest.     

The Effect of Hydrolysis with Neutrase on Molecular Weight,Functional Properties,and Antioxidant Activities of Alaska Pollock Protein Isolate

In this study, the Alaska pollock protein isolate(APPI) was hydrolyzed by Neutrase for 20, 40, 80, 120, 160, 200, and 240 min. Hydrolysates with different molecular weights were produced and they were named as H1–H7. Furthermore, the effects of hydrolysis on the average molecular weights, functional properties(solubility, oil-holding capacities, foaming activities, and emulsifying properties), and antioxidant activities(1, 1-diphenyl-2-picrylhydrazyl, superoxide, and hydroxyl free radical-scavenging activities) were determined. It was found that when the degree of hydrolysis(DH) increased, the average molecular weights of the hydrolysates decreased significantly. The functional properties of APPI were also significantly improved. The hydrolysates of APPI exhibited better solubility, emulsifying activities, and foaming activities. Hydrolysates with low molecular weights(1 kDa) had better solubility, oil-holding capacities, and emulsifying activities, while hydrolysates with higher molecular weights(1 kDa) had better foaming activities. In addition, the hydrolysates exhibited excellent antioxidant properties, while the inhibition values of 1, 1-diphenyl-2-picryl hydroxyl(DPPH), superoxide, and hydroxyl free radical-scavenging activities, were 85.22%, 53.56%, and 75.00% respectively, when the concentration of the hydrolysates was 5.0 mg mL~(-1). The lower the average molecular weight was, the higher was the antioxidant activity. These results indicated that hydrolysis with Neutrase is an effective method for improving the functional and antioxidant properties of APPI. The hydrolysates of APPI displayed great potentials to be used as natural antioxidants in protein-rich aqueous foods such as nutrient supplements and sports beverages.     

罗非鱼下脚料自溶条件的初步探讨

采用单因素和正交优化实验L9(43)确定酶法自溶罗非鱼下脚料的最适条件,结果表明,最适自溶条件为:料液质量比1∶3,反应温度50℃,pH7.5,反应时间6.5h;酶解结束蛋白质回收率为81.75%。     

Biochemical Composition and Nutritional Value of Different Shell Color Strains of Pacific Oyster <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

The shell color of Pacific oyster (Crassostrea gigas) is a desirable trait; but the nutritional studies on C. gigas with different shell colors have not been conducted. Through successive selective breeding, five shell color strains of black (B), purple (P), orange (O), golden (G) and white (W) C. gigas have been developed. The aim of this study was to evaluate the chemical composition and nutritional value of five shell color strains and one commercial population with a common color. The biochemical composition including moisture, total protein, glycogen, ash, total fat, fatty acids (FA), amino acids and minerals was detected. The results indicated that the protein (50.76%–56.57%) was the major component. The content of glycogen showed a significant difference between orange shell and golden shell strains, as well as between commercial population and golden shell strain. In addition, all shell color strains contained a large amount of essential amino acids (12.20–14.15 g (100 g)^?1), of them leucine (2.81–3.29 g (100 g)^?1) and lysine (2.79–3.28 g (100 g)^?1) were predominant. The oysters were rich in polyunsaturated fatty acids (42.26%–45.24% of total fatty acid) with high levels of DHA (18.53%–21.16% of total fatty acid) and EPA (17.23%–18.68% of total fatty acid). Significant differences of mineral contents (Mg, Zn, Fe and Cu) were identified among the six populations. These results indicated that C. gigas with different shell colors presented rich nutritional value with high protein, glycogen, essential amino acids and polyunsaturated fatty acids. The biochemical composition obtained in this study is useful for selective breeding of C. gigas with different shell colors.     

Construction and characterization of a normalized cDNA library of Nannochloropsis oculata (Eustigmatophyceae)

We constructed and characterized a normalized cDNA library of Nannochloropsis oculata CS-179, and obtained 905 nonredundant sequences (NRSs) ranging from 431–1 756 bp in length. Among them, 496 were very similar to nonredundant ones in the GenBank (E ≤1.0e-05), and 349 ESTs had significant hits with the clusters of eukaryotic orthologous groups (KOG). Bases G and/or C at the third position of codons of 14 amino acid residues suggested a strong bias in the conserved domain of 362 NRSs (>60%). We also identified the unigenes encoding phosphorus and nitrogen transporters, suggesting that N. oculata could efficiently transport and metabolize phosphorus and nitrogen, and recognized the unigenes that involved in biosynthesis and storage of both fatty acids and polyunsaturated fatty acids (PUFAs), which will facilitate the demonstration of eicosapentaenoic acid (EPA) biosynthesis pathway of N. oculata. In comparison with the original cDNA library, the normalized library significantly increased the efficiencies of random sequencing and rarely expressed genes discovering, and decreased the frequency of abundant gene sequences.     

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.     

Monthly Changes of Glycogen, Lipid and Free Amino Acid of Oyster

Monthly difference of the chemical composition of oyster cultured along the eastern coast of Shandong Province was analyzed. The components analyzed included glycogen, fatty acid and free amino acid （FAA）. The content of glycogen was high in January and March （2.89 and 2.82 g（100 g） ^-1 on average, respectively） and low in October （2.07g（100g）^- 1 on average）. The low content of neutral lipids in October reflected a relatively poor nutritional value of oyster （1.42 g（ 100 g）^- 1 on average）. The main fatty acids of oyster were palmitic acid （16：0）, oleic acid （18： 1）, eicosapentaenoic acid （EPA, 20： 5（0 3） and docosahexaenoic acid （DHA, 22：6（0 3）. The major FAAs of oyster were Taurine, Glutamicacid, Glycin, Alanine, Arginine and Proline. Taurine was the most abundant FAA with its content ranging from 603 mg（100 g） ^-1 to 1 139 mg（ 100 g） ^-1. The high contents of glycogen, polyunsaturated fatty acid and FAA showed that oyster cultured along the eastern coast of Shandong Province was nutritionally good in January and March.     

Preparation by enzymolysis and bioactivity of iron complex of fish protein hydrolysate （Fe-FPH）from low value fish

Preparation of Fe^2＋ chelate of fish protein hydrolysate （Fe-FPH） obtained from low value fish proteins was introduced and its bioactivity was studied by compound enzymolysis. The optimum conditions for hydrolysate chelating Fe^2＋ are DH （degree of hydrolysis） at 5%, pH 7.0, 20℃ and 15 min chelating time for FM （material not being defatted）. Four types of Fe-FPH including CA （deposit after chelating）, CB （deposit in 50% of absolute ethanol solution）, CC （suspended deposit in 80% of absolute ethanol solution）, and CD （bottom deposit in 80% of absolute ethanol solution） were fractionated with absolute ethanol from FM. Structural analysis through infra-red spectrum revealed that Fe^2＋ was combined strongly with amino-group and carboxyl-group in each chelate and each Fe^2＋ could form two five-member ring structures. All of the four chelates were shown more significant antioxidative activity and can be used as natural hydrophobic and hydrophilic antioxidant. Among all the chelates, the CB possesses the most effective antioxidative activity at 92% as high as that of a-tocopherol. Among all Fe-FPHs, only CD showed the most effective antibacterial activity against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Bacillus subtilis and can be used as natural antibacterial. It provides a more effective way for utilization of low value fish proteins and key information of Fe-FPH as additive in food industry.     

4种斑节对虾亲虾饵料蛋白质的营养价值评价

分析了沙蚕、缢蛏、拖鱿鱼胴体、花蟹蟹肉以及斑节对虾亲虾肌肉和成熟卵巢基本营养成分、氨基酸组成。通过计算4种饵料与斑节对虾亲虾肌肉和成熟卵巢的必需氨基酸比率(A/E)的比值和必需氨基酸指数(EAAI),评价4种斑节对虾亲虾饵料蛋白质的营养价值。结果表明:沙蚕、缢蛏、拖鱿鱼胴体、花蟹蟹肉以及斑节对虾(Penaeusmonodon)亲虾肌肉和成熟卵巢中的粗蛋白质量分数分别为9.18%、8.14%、12.10%、17.00%、21.90%、18.60%;总氨基酸质量分数分别为7.30%、7.47%、11.70%、14.80%、19.40%、14.60%。以斑节对虾亲虾肌肉蛋白为参比,沙蚕、缢蛏、拖鱿鱼胴体以及花蟹蟹肉的必需氨基酸指数分别0.93、0.94、0.94、0.97;以斑节对虾亲虾成熟卵巢蛋白为参比,沙蚕、缢蛏、拖鱿鱼胴体以及花蟹蟹肉的必需氨基酸指数分别0.95、0.93、0.88、0.84。这说明4种亲虾饵料相对斑节对虾亲虾而言是一种优质蛋白质,能够满足亲虾蛋白需求。不过,精氨酸、组氨酸等必需氨基酸,不能完全满足亲虾需求。     

Metabolic plasticity of nitrogen assimilation by Porphyra umbilicalis (Linnaeus) Kützing

The physical stresses associated with emersion have long been considered major factors determining the vertical zonation of intertidal seaweeds.We examined Porphyra umbilicalis(Linnaeus) Kützing thalli from the vertical extremes in elevation of an intertidal population(i.e.upper and lower intertidal zones) to determine whether Porphyra thalli acclimate to different vertical elevations on the shore with different patterns of nitrate uptake and nitrate reductase(NR) and glutamine synthetase(GS) activities in response to different degrees of emersion stress.We found that the nitrate uptake and NR recovery in the emersed tissues took longer in lower intertidal sub-population than in upper intertidal sub-population;and GS activity was also significantly affected by emersion and,interestingly,such an activity was enhanced by emersion of thalli from both upper and lower intertidal zones.These results suggested that intra-population variability in post-emersion recovery of physiological functions such as nutrient uptake and NR activity enables local adaptation and contributes to the wide vertical distribution of P.umbilicalis.The high GS activity during periodic emersion stress may be a protective mechanism enabling P.umbilicalis to assimilate nitrogen quickly when it again becomes available,and may also be an evidence of photorespiration during emersion.     

Cloning and Characterization of an Abalone （Haliotis discus hannai ） Actin Gene

1　Introduction　Actinisanimportantcontractileproteinineukary oticcells .Itisoneofthetwomajorcomponentsin volvedinthecontractionofmusclecells .Innon mus clecells ,itisthemajorpartofcytoskeletoninvolvedinmanyprocessessuchascellmotility ,endocytosis ,exocyt…     

Standardization of CPUE for Chilean jack mackerel (Trachurus murphyi) from Chinese trawl fleets in the high seas of the Southeast Pacific Ocean

The generalized linear model (GLM) and generalized additive model (GAM) were applied to the standardization of catch per unit effort (CPUE) for Chilean jack mackerel from Chinese factory trawl fishing fleets in the Southeast Pacific Ocean from 2001 to 2010 by removing the operational, environmental, spatial and temporal impacts. A total of 9 factors were selected to build the GLM and GAM, i.e., Year, Month, Vessel, La Niña and El Niño events (ELE), Latitude, Longitude, Sea surface temperature (SST), SST anomaly (SSTA), Nino3.4 index and an interaction term between Longitude and Latitude. The first 5 factors were significant components in the GLM, which in combination explained 27.34% of the total variance in nominal CPUE. In the stepwise GAM, all factors explained 30.78% of the total variance, with Month, Year and Vessel as the main factors influencing CPUE. The higher CPUE occurred during the period April to July at a SST range of 12–15°C and a SSTA range of 0.2–1.0°C. The CPUE was significantly higher in normal years compared with that in La Niña and El Niño years. The abundance of Chilean jack mackerel declined during 2001 and 2010, with an increase in 2007. This work provided the relative abundance index of Chilean jack mackerel for stock assessment by standardizing catch and effort data of Chinese trawl fisheries and examined the influence of temporal, spatial, environmental and fisheries operational factors on Chilean jack mackerel CPUE.     

Measurement of trace manganese (II) by the catalytic kinetic spectrophotometric method

A new kinetic spectrophotometric method is developed for the measurement of manganese (II) in water. The method is based on the catalytic effect of manganese (II) with the oxidation of weak acid brilliant blue dye (RAWL) by KIO₄ using the Nitrilo triacetic acid (NTA) as an activation reagent. The optimum conditions obtained are 40 mgL⁻¹ RAWL, 1×10⁻⁴molL⁻¹ KIO₄, 2×10⁻⁴ molL⁻¹ Nitrilo triacetic acid (NTA), pH = 5.8, the reaction time of 3.00 min and the temperature of 20.0 °C. Under the optimum conditions, the proposed method allows the measurement of manganese (II) in a range of 0–50.0 ng mL⁻¹ and with a detection limit of down to 0.158 ng mL⁻¹. The recovery efficiency in measuring the standard manganese (II) solution is in a range of 98.5%–102%, and the RSD is in a range of 0.76%–1.25%. The new method has been successfully applied to the measurement of manganese (II) in both fresh water and seawater samples with satisfying results. Moreover, few cations and anions interfere with the measurement of manganese (II). Compared with other kinetic catalytic methods and instrumental methods, the proposed method shows fairly good selectivity and sensitivity, low cost, cheapness, low detection limit and rapidity. It can be applied on boats easily.     

Comparative Analysis of Nutrient Composition of Caulerpa lentillifera from Different Regions

This study investigated the difference of nutrient composition in Caulerpa lentillifera collected from different regions.The nutrient compositions of C.lentillifera from China's Hainan and Shandong provinces were determined and compared with those from Semporna(Malaysia),Sabah(Malaysia),Petchburi(Thailand),and two other species of seaweeds(Malaysia).The results showed that the polysaccharide and protein contents of C.lentillifera from Hainan(44.82%±0.98%and 12.5%±0.70%),Shandong(43.22%±1.42%and 14.7%±0.72%),Petchburi(59.27%±2.31%and 12.49%±0.30%),and Sabah(44.02%±2.01%and 19.38%±1.48%)were higher than those of Eucheuma cottonii(26.49%±3.01%and 9.76%±1.33%)and Sargassum polycystum(33.49%±1.70%and 5.40%±0.07%),respectively.The polyunsaturated fatty acid contents in C.lentillifera from Hainan(29.98%)and Shandong(22.11%)were higher than that in Semporna(16.76%±0.27%),Sabah(9.49%),and S.polycystum(20.34%±0.43%),but lower than that in E.cottonii(51.55%±0.57%).In Caulerpa lentillifera of Hainan and Shandong provinces,the essential amino acid(EAA)/total amino acid(TAA)ratios were 35.22%and 38.78%,respectively;and the EAA/ETAA ratios were 54.36%and 63.35%,respectively.The EAA composition of C.lentillifera was close to the ideal model of Food and Agriculture Organization/World Health Organization.C.lentillifera was rich in mineral elements,including calcium((3315.85±127.55)mg(100 g)^-1 and(3728.35±92.38)mg(100 g)^-1),magnesium((6715.74±82.58)mg(100 g)^-1 and(8128.59±242.72)mg(100 g)^-1),and trace elements,including iron((510.65±5.47)mg(100 g)^-1 and(1972.97±183.35)mg(100 g)^-1),selenium((1.04±0.08)mg(100 g)^-1 and(0.83±0.15)mg(100 g)^-1),and zinc((33.90±0.13)mg(100 g)^-1 and(11.75±2.05)mg(100 g)^-1).These results showed that C.lentillifera was more nutritious than S.polycystum or E.cottonii in terms of polysaccharide,protein,and fatty acid contents.Furthermore,both C.lentillifera species collected from Hainan and Shandong provinces show more basic nutrients.Therefore,C.lentillifera has important development and application prospects.