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.     

龙虎斑鱼肌肉营养成分分析

对龙虎斑鱼的肌肉进行营养成分分析和肉质评价,结果表明,龙虎斑鱼新鲜肌肉中粗脂肪质量分数4.18%,粗蛋白质量分数17.05%,粗灰分质量分数1.24%,水分质量分数80.19%.肌肉蛋白质中共检测出氨基酸种类有17 种,占新鲜肌肉样品的质量分数为17.88%;7 种必需氨基酸（EAA）占新鲜肌肉样品的质量分数为7.79%,占氨基酸总量的43.57%;4 种鲜味氨基酸（DAA）占新鲜肌肉样品的质量分数为6.17%,占氨基酸总量的34.51%;必需氨基酸指数（EAAI）为101.35,其氨基酸组成比例符合FAO/WHO 标准.新鲜肌肉中共检测出脂肪酸种类有14 种.其中7 种饱和脂肪酸（SFA）占总脂肪酸的质量分数为47.29%;4 种单不饱和脂肪酸（MUFA）占总脂肪酸的质量分数为18.86%;3 种多不饱和脂肪酸（PUFA）占总脂肪酸的质量分数为33.86%,DHA 质量分数占脂肪酸总量的30.24%.龙虎斑鱼营养价值较高,海鲜味浓郁.     

Effects of polypeptide from<Emphasis Type="Italic">Chlamys farreri</Emphasis> on amino acid content in guinea pig skin irradiated by chronic ultraviolet A and B

We examined the effects of polypeptide fromChlamys farreri (PCF) on the amount of hydroxyproline in guinea pig skin irradiated by chronic ultraviolet A (UVA) and ultraviolet B (UVB) radiation. PCF was applied locally before repeated exposure of guinea pig to UVA and UVB. The contents of hydroxyproline and other amino acids in guinea pig skin were determined by automatic amino acid analyzer. Our results showed that: (1) long-time UVA and UVB radiation can reduce dramatically the amounts of hydroxyproline, aspartic acid, threonine, glycine, phenylalanine and lysine in guinea pig skin in comparison with the control group (P<0.05); (2) Compared with model group, pre-treatment with 5% and 20% PCF prior to UVA and UVB radiation can inhibit the decline of amino acids content in guinea pig skin in a dose-dependent manner (P<0.05). As the decrease of hydroxyproline, glycine and lysine contents in the skin directly reflexes type I collagen degeneration, our results indicated that the chronic application of PCF can protect skin type I collagen against UV radiation, and thus protect skin from photoaging. Project No. 39970638 supported by the NSFC, and also supported by the Science and Technology Bureau of Qingdao (No: 2001-28-50).     

Evalution on nutritive value ofPortunus trituberculatus

This study on the nutritive indexes (total nitrogen, amino acids, crude fats, inorganic elements, unsaturated fatty acids) of meat, male and female reproductive gland ofPortunus trituberculatus showed that their nutritive value is in the order meat>female reproductive gland>male reproductive gland and that they do not raise the total cholesterol (TC) and triglyceride (TG) contents of the serum of the animals which eat them but increase the contents of high density lipoprotein cholesterol (HDL-C) in the animal's blood. These findings implied that people who have high blood lipid and aortic atheroma can safely use them as food. This study showed thatP. trituberculatus has high nutritive value.     

饲粮添加人参对贵妃鸡肌肉氨基酸水平和血液生理生化指标的影响

选用90日龄的贵妃公鸡随机分成4组,在基础日粮中按质量分数0.00%﹑0.02%﹑0.04%和0.08%添加人参,实验期45 d,探讨人参对贵妃鸡肌肉氨基酸含量和血液生理生化指标影响。结果表明:0.04%的人参添加量使得腿肌总氨基酸量和鲜味氨基酸含量比对照组分别提高19.19%和14.81%;0.04%的人参添加量使得胸肌总氨基酸含量和必需氨基酸含量比对照组分别提高18.84%和16.54%,鲜味氨基酸含量比对照组提高了14.82%。日粮中添加0.04%的人参能显著提高贵妃鸡胸肌和腿肌的苏氨酸﹑蛋氨酸和缬氨酸的含量。各实验组血清谷草转氨酶和谷丙转氨酶的活性均低于对照组,差异具有统计学意义(P0.05)。饲粮中添加0.04%人参添加实验剂效果最佳。     

DETERMINATION OF AMINO ACIDS IN TWO POLYSIPHONIA SPECIES AND STUDY OF ENZYMATIC HYDROLYSIS METHOD

The total content ot the nch amino acids in two common red algae, Potystpnoma urceolata and Polysiphonia japonica growing in the Qingdao seashore were determined. The algae powder was hydrolyzed by 6 mol/L HC1 at 110℃ for 48 h and determined by amino acid analyzer. The content was 25.35% and 24.16% , respectively, much higher than that of some other species. In addition, a nutritive liquid with abundant amino acids was prepared (by the enzymatic hydrolysis method using Polysiphonia urceolata ) as raw material for a kind of health beverage. The dried seaweed was decolored by 0.25% KMnO4 and 0.5% active carbon, then enzymalized. In the selection of enzymalizing condition, the orthogonal experimental design was used. Four factors including kinds of enzyme, quantity of enzyme, temperature and time were studied at 3 levels. According to the orthogonal design results, we can choose an optimal condition: hydrolyzing at 45℃ by neutral proteinase (0.25% , w/w) for 2 h, adjusting pH to 8.5, then adding trypsin (0.25% , w/w) and hydrolyzing for 2 h. Finally the above solution was alkalized by NaOH and neutralized by casein. After the hydrolyzed liquid was filtered and concentrated, suitable additives were added. The final products contain rich amino acids.     

Vertical variations and composition of dissolved free amino acid in the seawater of the Yap Trench in the western Pacific Ocean

The composition and concentration of dissolved free amino acid (DFAA) of seawater samples collected in May 2016 from the surface to the hadal zone of the northern region of the Yap Trench were analyzed by pre-column derivatization of o-phthalaldehyde. Results show that the average concentration of DFAA in the study area was 0.47±0.36 µmol/L. In different sampling stations, the concentrations of DFAA with water depth showed complex variation patterns. At the sediment-seawater interface, the concentrations of DFAA in the western side of the trench were obviously higher than that in its eastern side. In the study area, there were no significant correlations between the concentrations of DFAA and the environmental parameters such as concentrations of chlorophyll a (Chl a), dissolved oxygen (DO), pH, and dissolved inorganic nitrogen (DIN), indicating that the concentrations of DFAA in seawater of the trench are affected by many factors, such as photosynthesis, respiration, temperature, pressure, illumination, and circulation. The dominant DFAA are similar in different water layers of sampling stations, including aspartic acid (Asp), glutamic acid (Glu), glycine (Gly), and serine (Ser). The composition of different amino acids, and the relative abundance of acidic, basic, and neutral amino acids might be related to the sources and consumption of various amino acids. Nine pairs of amino acids in the DFAA showed significantly positive relationship by correlation matrix analysis, suggesting that they might share similar biogeochemical processes. The degradation index (DI) of the DFAA in seawater of the Yap Trench could reflect the degradation, source, and freshness of DFAA in the trench to some extents. This is a preliminary study of amino acids from sea surface to hadal zone in the ocean, more works shall be done in different trenches to reveal their biogeochemical characteristics in extreme marine environments.

     

罗氏沼虾三群体间肌肉营养品质的比较

对饲养在相似环境下的罗氏沼虾(Macrobrachium rosenbergii)三群体肌肉营养成分进行了分析和比较,3个群体分别为浙江湖州的人工养殖群体(A)、缅甸引进种子二代群体(B)和“南太湖1号”杂交群体(B-+)。结果表明,3个群体粗蛋白含量的高低顺序依次为:B-+群体(17.57%)、B群体(16.65%)、A群体(15.88%),但三群体之间没有显著性差异(P>0.05);而A群体的粗脂肪含量为2.04%,显著高于B群体的1.24%(P<0.05)。3个群体中,B-+群体的总氨基酸含量、必需氨基酸含量、鲜味氨基酸和脂肪酸中的EPA+DHA含量均为最高,且必需氨基酸指数最高,为68.06;相比之下,A群体的上述各项指标值均较低。可见,以上三群体中杂交种群(B-+)的肌肉营养品质较优,而人工养殖群体(A)的肌肉品质相对较差。     

The selection of alkaline protease-producing yeasts fi＇om marine environments and evaluation of their bioactive peptide production

A total of 400 yeast strains from seawater, sediments, saltern mud, marine fish guts, and marine algae were obtained. The protease activity of the yeast cultures was estimated, after which four strains (HN3.11, N11b, YF04C and HN4.9) capable of secreting extracellular alkaline protease were isolated. The isolated strains were identified as Aureobasidium pullulans, Yarrowia lipolytica, Issatchenkia orientalis and Cryptococcus cf. aureus. The optimal pH of the protease activity produced by strains HN3.11, YF04C, and HN4.9 was 9.0, while that of the protease produced by strain N11b was 10.0. The optimal temperature for protease activity was 45°C for strains HN3.11, N11b, and YF04C, and 50°C for strain HN4.9. After digestion of shrimp (Penaeus vannamei) protein and spirulina (Arthospira platensis) protein with the four crude alkaline proteases, the filtrate from spirulina (Arthrospira platensis) powder digested by the crude alkaline protease of strain HN3.11 was found to have the highest antioxidant activity (61.4%) and the highest angiotensin I converting enzyme (ACE)-inhibitory activities (68.4%). The other filtrates had much lower antioxidant activity and ACE-inhibitory activities.     

米曲霉中性蛋白酶特性的研究

研究了米曲霉中性蛋白酶的特性。结果表明,米曲霉中性蛋白酶的最佳提取方法为加入pH 7.2的缓冲液研匀,置恒温(30℃)培养箱中提取30 min,每10 min搅动1次,最后用4层纱布过滤。酶反应最适温度为55℃,最适pH值为7.2。NaCl和EDTA对其有一定的抑制作用。酶的热稳定性好,在45℃的温度下处理24 h后,酶活力仍有45.5%。米曲霉中性蛋白酶与培养物一起保存时,稳定性也较好,在4℃保存一个月后,酶活力几乎无变化。     

The effects of nitrogen,phosphorus, vitamins and trace metals on the growth of the red tide organismProrocentrum Micans

Laboratory culture experiments showed that <100μ mol/L nitrate, amonium or mixture of amino acids promote the growth of the red tide organismProrocentrum micans Ehrenb, but that >100μmol/L of ammonium, or mixture of glycine and glutamate was harmful to growth, and that orthophosphate wasP. micans’ main phosphorous source in the ocean. Presence of 80μ mol/L EDTA, 0.5 to 1 μmol/L Fe³⁺, 1.0 to 20.0 μ mol/L Mn²⁺ 0.1 to 0.4 μmol/L Co²⁺ in the culture medium could improve the growth ofP. micans. Vitamin B₁ promoted growth, but vitamin B₁₂ and biotin did not. The estimated minimum cell quotas (q _o) for nitrogen and phosphorus being 0.74 pmole/cell and 0.045 pmole/cell show that phosphorus (more than nitrogen) limits the growth ofP. micans in the study area. This project was supported by the Natural Science Foundation of Zhejiang Province.     

Purification and characterization of an alkaline protease from Acetes chinensis

An alkaline protease from Acetes chinensis was purified and characterized in this study. The steps of purification include ammonium sulfate precipitation, ion-exchange chromatography with Q-sepharose Fast Flow, gel filtration chromatography with S300 and the second ion-exchange chromatography with Q-sepharose Fast Flow. The protease was isolated and purified, which was present and active on protein substrates (azocasein and casein). The specific protease activity was 17.15 folds and the recovery was 4.67. The molecular weight of the protease was estimated at 23.2 kD by SDS-PAGE. With azocasein as the susbstrate, the optimal temperature was 55°C and the optimal pH value was 5.5. Ion Ca²⁺ could enhance the proteolytic activity of the protease, while Cu²⁺, EDTA and PMSF could inhibit its activity.     

Proteocephalus sinocyclocheili n. sp. (Cestoda: Proteocephalidae) in a fishSinocyclocheilus grahami tingi Fang from Yunnan, China

A new species ofProteocephalus from the fishSinocyclocheilus grahami tingi, a subspecies endemic to Fuxian Lake in Southwest China,is described. This is the first report of cestode from this host. The new species differs fromP. exiguus, P. Longicollis, P. parasiluri andP. torulosus, the only other species ofProteocephalus currently reported from China, by lacking an apical sucker, fromP. exiguus by having 45–108 rather than 24–54 testes per proglottis, and a maximum of 15 rather than 28 uterine diverticula; fromP.longicollis by having an average of 45–108 rather than 75–115 testes per proglottis and testes being in a single layer instead of 2 layers; and fromP. torulosus by having 45–108 instead of 150 testes per proglottis and testes being in 3–4 layers; and fromP. parasiluri by having 45–108 rather than 180 testes per proglottis. It resemblesP. percae, P. pollanicola, andP. thymalli in number of testes and uterine diverticula, but differs by lacking an apical sucker.Proteocephalus thymalli, like the new species, has a vagina that opens dorsally to the cirrus sac, butP. thymalli slao has postovarian testes, which the new species lacks. The possible significance of parasitological examinations of the other two subspecies ofS. grahami in two lakes close to Fuxian Lake is discussed in relation to host speciation and coevolution of the host-parasite system.     

Comparative study of the nutritional composition and toxic elements of farmed and wild Chanodichthys mongolicus

Information of the difference in quality between farmed and wild fish is central to better ensuring fish products produced in aquaculture meet regulatory and consumer requirements.Proximate composition,amino acid and fatty acid profiles,and toxic elements contents of farmed and wild Chanodichthys mongolicus were established and compared.Significantly higher crude protein content while lower moisture content in farmed fish compared to wild fish were observed(P0.05).The percentages of total amino acids(TAA),total essential amino acids(TEAA),total non-essential amino acids(TNEAA)and total delicious amino acids(TDAA) in farmed fish were all significantly higher than those in the wild equivalent(PO.05).Compared to the FAO/WHO recommended reference values,the ratios of TEAA/TAA(39.84%-40.33%) were comparable to 40%and TEAA/TNEAA(66.22%-67.60%) were above 60%.Fatty acid profiles in both fanned and wild C.mongolicus were dominated by monounsaturated fatty acid(MUFA),with farmed fish contained much more MUFA content compared to wild counterpart(P0.05).Notably,wild fish exhibited significantly higher levels of total polyunsaturated fatty acid(PUFA) including eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) than farmed fish(P0.05).The EPA(C_(20:5n3))and linoleic acid(C_(18:2n6)) were the predominant PUFA in wild and fanned C.mongolicus,respectively.Moreover,farmed fish displayed an overall lower toxic element levels(As,Cd,Pb and Hg) in comparison with wild fish,and both were far lower than the established limit standard.In conclusion,our results suggest that the nutritional quality of farmed C.mongolicus was inferior to their wild counterpart with respect to fatty acids nutrition,and therefore further studies should focus on the improving C.mongolicus diet in order to enhance the overall nutritional composition.     

SIUDIES ON THE BIOCHEMICAL COMPOSITION,ENERGETICS AND ESSENTIAL AMINO ACIDS OF THREE MUDSKIPPERS IN XIAMEN HARBOUR

Measurements were made on the contents of protein,lipid,glycogen(PLG)and water,and on caloric values and amino acids,in muscle of three mudskippers periophthalmus cantonensis,Scarteiaosviridis and Boleophthalmus pectinirostris collected from Haicang,Xiamen.The essential amino acids(EAA)for these fishes were also studied with radioisotopic trace method.The results showed:(1)Thecontent of each component in tested fish muscles differed slightly,and protein was the most importantcomponent making up from 6.685% to 9.891% of the wet weight (about 44.21%-50.45% of dryweight);(2)Energy calculated from the sum of protein, lipid and glycogen in wet muscle was low (<4.3kJ/g) in these Fishes, especially in B. pectinirostris(<3.1 kJ/g), the ratios of energy to protein content (E/P) also were low(<39.873-45.535kJ/g), (3)Seventeen amino acids were determined in thesethree fishes.The content of the same amino acid(among the seventeen) tested in different species andsexes varied slightly.The amounts of methionine, phenyl     

The Complete Sequence of Mitochondrial COⅡ Gene of Fenneropenaeus chinensis and Its Applicability as a Marker for Phylogenetic Analysis

The complete mitochondrial cytochrome oxidase subunit Ⅱ （COⅡ） gene of Penaeinae shrimp Fenneropenaeus chinensis was cloned and sequenced. The gene is 688 bp in length and codes for 229 amino acids. It shows 83.2%, 87.0% and 83.8% sequence similarity to Marsupenaeus Japonicus, Penaeus monodon and Farfantepenaeus notialis, respectively. The A＋T content of the whole gene and that at the third position of codons are 64.7% and 78.2%, respectively. The phylogenetic relationship between F. chinensis and three other species representing genera Farfanatepenaeus, Marsupenaeus and Penaeus was analyzed. Results showed that the genetic distances among the four taxa ranged from 0.144 0 to 0.200 5, exceeding those estimated with COⅠ and partial 16S rRNA gene sequences among Marsupenaeus, Litopenaeus and Melicertus, and being therefore larger than the value among subgenera. It has been suggested that the COⅡ gene has a faster evolutionary rate than that of the COⅠ gene and partial 16S rRNA gene and could be used for phylogenetic analysis at genus or species level. The results of the present study indicated that Farfantepenaeus, Fenneropenaeus, Marsupenaeus and Penaeus are at a higher phylogenetic level than subgenus, which supports the opinion of the elevation of phylogenetic status of the four subgenera to genus level.     

Isolation and characterization of a marine bacterium producing protease from Chukchi Sea, Arctic

A Gram negative bacterium Ar/W/b/75°25＇N/1 producing extracellular alkaline protease was isolated from surface water of latitude 75°25＇N, and longitude 162°25＇W in Chukchi sea, Arctic. The strain can grow at the temperature range from 7℃ to 30℃, and grow better at 30(℃. It can not grow at 40℃. Keeping certain salinity concentration in medium is necessary for cell growth. It grows well in medium containing salinity concentration from 0. 5 % to 10 % sodium chloride. Glucose, sucrose and soluble starch can be utilized by the strain, among which glucose is the optimal carbon source. Peptone is the optimal organic nitrogen source for cell growth and protease producing, and ammonium nitrate is the optimal inorganic nitrogen source.About 75.7% of total protease of the strain are extracellular enzyme. Optimal temperature for proteolytic activity is at 40℃. Protease of the strain keeps stable below 40℃, and shows high proteolytic activity within the pH range from 7 to 11.     

Screening and characterization of the alkaline protease isolated from PLI-1, a strain of Brevibacillus sp. collected from Indonesia’s hot springs

A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia’s hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70℃ and pH 8.0-9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL-1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70℃ and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.     

DABS-Cl柱前衍生高效液相色谱法测定对虾肌肉中18种游离氨基酸

建立一种以DABS-Cl为柱前衍生剂,利用反相高效液相色谱测定对虾肌肉中18种游离氨基酸的方法。结果表明,在50 min内18种氨基酸得到完全分离,各氨基酸相关系数均大于0.997,氨基酸的回收率在86.60%~107.27%之间(胱氨酸除外),方法的检出限为0.18~0.66 mg/L,相对标准偏差为3.53%~8.51%(天冬氨酸除外)。