Pufferfish are considered a culinary delicacy but require careful preparation to avoid ingestion of the highly toxic tetrodotoxin (TTX), which accumulates in certain tissues. In this study, the tissue distribution of peroxiredoxin-1 from Takifugu bimaculatus was investigated. The peroxiredoxin-1 protein was obtained by in vitro recombinant expression and purification. The recombinant protein had a strong ability to scavenge hydroxyl radicals, protect superhelical DNA plasmids from oxidative damage, and protect L929 cells from H2O2 toxicity through in vitro antioxidant activity. In addition, we verified its ability to bind to tetrodotoxin using surface plasmon resonance techniques. Further, recombinant proteins were found to facilitate the entry of tetrodotoxin into cells. Through these analyses, we identified, for the first time, peroxiredoxin-1 protein from Takifugu bimaculatus as a potential novel tetrodotoxin-binding protein. Our findings provide a basis for further exploration of the application of peroxiredoxin-1 protein and the molecular mechanisms of tetrodotoxin enrichment in pufferfish.

Extracting peptides from fish bones as calcium supplements is of great significance for patients with osteoporosis. Novel peptides were obtained from Muraenesox cinereus bone (MCBP) by enzymatic hydrolysis and membrane separation. Molecular dynamics simulation and electron microscope analysis shown MCBP could chelate with calcium ions. A Caco-2 cell monolayer model was used to investigate the effect of peptide–calcium chelates on promoting calcium transport. MC3T3-E1 osteoblasts were treated with different concentrations of MCBP (0, 0.25, 0.5, or 1 mg/mL) to analyze the effects of MCBP on calcium absorption and cell differentiation factors. The bone formation of zebrafish evaluate the therapeutic effect of MCBP on the zebrafish OP model. The results showed that there were at least 13 kinds of calcium chelation modes in the mixed peptide, and peptide–calcium chelate could significantly improve calcium transport in the Caco-2 cell monolayer. After 14 days of culture with different concentrations of MCBP, the calcium deposition in MC3T3-E1 cells increased significantly. The type I collagen (COL-I), osteocalcin (OCN) and run-related transcription factor 2 (RUNX2), and activities of alkaline phosphatase (ALP) were dose-dependent with MCBP. MCBP can increase bone mineral density in an OP model of zebrafish. After being treated with MCBP, the expression levels of ALP, OCN, osteoprotegerin (OPG) and transforming growth factor-β (TGF-β) in the model group increased, while the expression levels of tartrate-resistant acid phosphatase (TRAP) decreased. The results indicated that MCBP may act as a nutritional food and provide an effective means for dietary supplementation in OP patients.

Alcalase, neutral protease, and pepsin were used to hydrolyze the skin of Takifugu flavidus. The T. flavidus hydrolysates (TFHs) with the maximum degree of hydrolysis (DH) and angiotensin-I-converting enzyme (ACE)-inhibitory activity were selected and then ultra-filtered to obtain fractions with components of different molecular weights (MWs) (<1, 1–3, 3–10, 10–50, and >50 kDa). The components with MWs < 1 kDa showed the strongest ACE-inhibitory activity with a half-maximal inhibitory concentration (IC50) of 0.58 mg/mL. Purification and identification using semi-preparative liquid chromatography, Sephadex G-15 gel chromatography, RP-HPLC, and LC–MS/MS yielded one new potential ACE-inhibitory peptide, PPLLFAAL (non-competitive suppression mode; IC50 of 28 μmmol·L−1). Molecular docking and molecular dynamics simulations indicated that the peptides should bind well to ACE and interact with amino acid residues and the zinc ion at the ACE active site. Furthermore, a short-term assay of antihypertensive activity in spontaneously hypertensive rats (SHRs) revealed that PPLLFAAL could significantly decrease the systolic blood pressure (SBP) and diastolic blood pressure (DBP) of SHRs after intravenous administration. These results suggested that PPLLFAAL may have potential applications in functional foods or pharmaceuticals as an antihypertensive agent.

[目的]研究菊黄东方鲀(Takifugu flavidus)精巢的醇提物(AETT)对小鼠的抗疲劳作用及毒理安全性.[方法]配制不同剂量的菊黄东方鲀精巢醇提物溶液及对照组,通过动物实验,测定小鼠的负重游泳时间和血清尿素氮、血清乳酸、肝糖原含量的变化,并观察在一次性给予最大剂量的条件下小鼠的死亡情况.[结果]菊黄东方鲀精巢醇提物高剂量组能显著延长小鼠负重游泳时间(P<0.01),提高小鼠体内肝糖原的储备量(P<0.01),降低运动后血清乳酸含量(P<0.01),降低运动后血清尿素氮含量(P<0.01),加速体内尿素氮的清除,具有显著的抗疲劳作用.以15 g/kg剂量给予小鼠菊黄东方鲀精巢醇提物,小鼠无死亡,体质量、毛色、脏器及行为活动均正常,无毒副作用.[结论]菊黄东方鲀精巢醇提物具有明显的抗疲劳活性,且安全无毒.

The von Willebrand factor type D (VWD) domain in vitellogenin has recently been found to bind tetrodotoxin. The way in which this protein domain associates with tetrodotoxin and participates in transporting tetrodotoxin in vivo remains unclear. A cDNA fragment of the vitellogenin gene containing the VWD domain from pufferfish (Takifugu flavidus) (TfVWD) was cloned. Using in silico structural and docking analyses of the predicted protein, we determined that key amino acids (namely, Val115, ASP116, Val117, and Lys122) in TfVWD mediate its binding to tetrodotoxin, which was supported by in vitro surface plasmon resonance analysis. Moreover, incubating recombinant rTfVWD together with tetrodotoxin attenuated its toxicity in vivo, further supporting protein–toxin binding and indicating associated toxicity-neutralizing effects. Finally, the expression profiling of TfVWD across different tissues and developmental stages indicated that its distribution patterns mirrored those of tetrodotoxin, suggesting that TfVWD may be involved in tetrodotoxin transport in pufferfish. For the first time, this study reveals the amino acids that mediate the binding of TfVWD to tetrodotoxin and provides a basis for further exploration of the molecular mechanisms underlying the enrichment and transfer of tetrodotoxin in pufferfish.

为了研究牡蛎寡肽对自由基清除作用及对人肝细胞L02细胞氧化损伤的保护作用,测定了牡蛎寡肽对羟自由基(·OH)的清除率,并建立过氧化氢(hydrogen peroxide,H2O2)氧化损伤人肝细胞L02的模型,研究牡蛎寡肽对氧化损伤L02细胞的存活率、活性氧(Reactive Oxygen Species,ROS)、抗氧化酶系(superoxide dismutase/SOD,glutathione/GSH)含量的影响.结果 表明,牡蛎寡肽对羟自由基的IC50为0.38 mg/mL.2 mg/mL牡蛎寡肽对于L02细胞的增殖率仍为123.98％.模型组SOD和GSH水平分别降低了40％、64.87％,而ROS增加了1倍.当添加不同剂量牡蛎寡肽后,中剂量组细胞中SOD含量几乎恢复到正常组水平,GSH活性也提高了118.89％,ROS水平降到模型组的62.43％,说明牡蛎寡肽对L02细胞无毒,能降低氧化损伤的L02细胞内的ROS水平,显著提高SOD和GSH含量,对细胞起到保护作用.因此,牡蛎寡肽对H2O2致氧化损伤的人肝L02细胞具有保护作用,可能是通过清除自由基,保护细胞内抗氧化酶系活性,抑制ROS的过多积累来实现.

为探讨牡蛎提取物对小鼠化学性肝损伤的保护作用,研究建立酒精导致小鼠急性肝损伤模型,监测各剂量组小鼠体重,测定肝组织丙二醛(Malondialdehyde,MDA) 、还原型谷胱甘肽(Glutathione,GSH) 和甘油三酯(Triglycerides,TG) 的含量及观察牡蛎提取物对肝组织病理的影响.结果表明,牡蛎提取物各剂量组小鼠的体重与模型组和空白组无显著性差异(P>0.05);高剂量组的MDA、TG明显低于模型对照组,高剂量组的GSH明显高于模型对照组,均呈显著性差异(P<0.05).高剂量组小鼠肝脏脂肪变性的程度明显低于模型组,差异显著(P<0.05).从而说明,牡蛎提取物对小鼠急性化学性肝损伤具有辅助保护功效.

为研究河鲀鱼精蛋白的制备工艺,以菊黄东方鲀(Takifugu Flavidus)精巢为原料,以鱼精蛋白提取率为指标,通过单因素试验研究NaCl溶液的pH、硫酸浓度、硫酸用量、提取温度、提取时间以及乙醇用量等单因素对鱼精蛋白提取率的影响,在此基础上,采用响应面法优化鱼精蛋白的提取工艺条件.结果显示,采用响应面法优化获得河鲀鱼精蛋白的最佳提取工艺条件为:NaCl溶液pH 8.5,硫酸浓度0.6 mol/L,硫酸用量4.0倍沉淀体积,提取温度45℃,提取时间2.4 h,乙醇用量4.0倍沉淀体积,此时鱼精蛋白提取率达到19.07％.研究表明,响应面法对河鲀鱼精蛋白提取条件的优化方案合理可行,可为河鲀精巢的开发利用提供参考依据.

为了对红毛藻的营养价值进行全面的分析及评价.采用国家标准生化测定法检测红毛藻(Bangia fusco-purpurea)的营养成分.测得红毛藻干品的水分、灰分、粗脂肪、粗蛋白质以及粗纤维的质量分数分别为10.90％、10.00％、3.78％、39.42％和17.20％;检出含有18种氨基酸,必需氨基酸质量分数为总量的35.53％,鲜味和甜味氨基酸质量分数占总氨基酸的49.87％,谷氨酸的质量分数最高(4.79％);测得脂肪酸总质量分数为3.77％,其中ω-3系列的多不饱和脂肪EPA的质量分数最高,占总脂肪酸质量分数的39.26％;红毛藻中含有多种矿物质如钾、钠、磷、镁、锰、钙、铁等元素,宏量及微量元素里质量分数最高分别为钾(2 627.17 mg/hg)和铁元素(31.80mg/hg).研究表明,红毛藻为高蛋白质和膳食纤维、营养价值丰富的经济海藻,具有较好的开发前景.

The effect of oyster oligopeptide on regulation of immune function in cyclophosphamide （CTX)-induced immunosuppressive mice was evaluated.The indexes of immune organs such as liver and spleen,the transformation and propagation ratio of T lymphocyte,the activity of NK cell,the humoral immunity,macrophage phagocytosis,the level of cell cytokines and hemolysin were assessed.The experiment results revealed that compared with the model immunosuppressed group,oyster oligopeptide could significantly enhance the proliferation of spleen lymphocyte,NK cell activity,expurgation index k,devouring index a,ability of devouring neutral red,the level of TNF-α、IL-6 and hemolysin,and regulate the distribution of CD3+ 4T and CD3+ 8T spleen lymphocyte subsets.However,the influence of oyster oligopeptide on the immune organs was not significant.It was concluded that oyster oligopeptide can improve the cell immunity,humoral immunity and non-specific immune function of immunosuppressive mice induced by CTX,and it can regulate the immune function positively in mice.

To develop a better quality microencapsulation of Anthocidaris crassispina gonad oil,Anthocidaris crassispina gonad oil was microencapsulated with the method of spray drying by using n-octenylsuccinate-derivatized starch(HI-CAP100) and β-cyclodextrin as the principal wall materials. Using emulsion stability and embedding rate as evaluation indexes,the effects of wall material ratio,the concentration of emulsion and the addition amount of oil on the microcapsules were studied through single factor experiments.Based on single factor experiments,orthogonal method was used to optimize the process conditions,and the physicochemical properties and hygiene indicators were detected. The results showed that, the optimum conditions of Anthocidaris crassispina gonad oil micro-encapsulation were:the ratio of HI-CAP100 and β-cyclodextrin was 4: 2,the concentration of emulsion 17%,and the addition amount of oil 15%;after being emulsified with IKA homogenizer for 30 min and homogenizing with a cooking machine for 5 min,the inlet air temperature of spray dryer 180 ℃,outlet air temperature 80 ℃,and the sampling rate of 500 mL/h,under which,95. 7% of embedding rate and 96. 8% of emulsion stability were obtained. The moisture content in the Anthocidaris crassispina gonad oil microcapsule was 2. 82%,the solubility was 71. 94%,the angle of auto-flowing was 22.05°,the total number of colonies was 480 CFU/g,the number of coliforms was less than 30 MPN/100 g, mold and yeast were less than 10 CFU/ g,Salmonella,Shigella and Staphylococcus aureus were not detected. The physicochemical properties and safety and hygiene indicators of Anthocidaris crassispina gonad oil microcapsules are all in line with the industry standards.

通过走访调研福建省、 市、 县/区的渔业主管部门以及福建省水产加工企业,结合近年的渔业统计数据等资料,分析了福建省水产加工现状、 主要问题及发展思路,形成福建省水产加工及鱼、 甲壳类、 贝、 藻产业现状及发展思路等系列综述性论文.十二五期间,福建省捕捞水产品产量稳中有升;养殖水产品产量增加明显;实施了渔业转型升级政策,福建省水产品加工产业规模不断扩大和加工技术水平不断上升,水产品质量安全水平大大提高,但加工比例偏低、 地区发展不平衡、 加工技术创新不够等问题仍然突出;提出并阐明符合福建水产品加工发展方向的模式,以期为实现福建渔业经济新增长提供参考依据.

目的:优化仿刺参抗氧化多肽的酶解工艺,并研究其对过氧化氢(hydrogen peroxide,H2O2)诱导的人脐静脉内皮细胞株EA.hy926损伤的保护效应.方法:以酶解产物的水解度、体外DPPH自由基清除率为指标,筛选出最适蛋白酶;在单因素实验基础上,选取温度、加酶量、pH作为影响因子,以体外DPPH自由基清除率为响应值,结合响应面试验优化酶解工艺条件;进一步探讨酶解多肽体外抗氧化活性.以MTS法检测低、中、高剂量组的仿刺参抗氧化多肽对H2O2诱导的血管内皮细胞损伤的保护作用,以MDA含量、SOD活力测定细胞氧化及抗氧化水平.结果:动物蛋白水解酶为最适蛋白酶,酶解工艺优化条件为:料液比1:20 g/mL、酶解时间2h、酶解温度50℃、加酶量7000 U/g、pH7.5,该条件下制备的仿刺参多肽体外DPPH自由基清除率为68.81％,与模型预测值(68.35％),相对误差为1％,回归模型可靠.酶解多肽对DPPH自由基、羟自由基(·OH)、超氧阴离子自由基(O2-·)和ABTS自由基(ABTS+·)的半数抑制浓度IC50分别是9.01、0.63、10.89和20.53 mg/mL,说明其具有较好的体外抗氧化活性.选取200 μmol/L浓度H2O2建立细胞损伤模型,与H2O2组相比,中、高剂量组仿刺参抗氧化多肽能明显抑制H2O2诱导的血管内皮细胞氧化损伤,降低MDA含量,提高SOD活力.结论:采用动物蛋白水解酶酶解优化工艺制备的仿刺参抗氧化多肽对人脐静脉内皮细胞EA.hy926具有显著的保护作用并呈显著的剂量-效应关系.

高效氯氰菊酯在农业中应用广泛,其中水产品可以富集水系统中的高效氯氰菊酯,并通过食物链,影响人体的健康.本研究通过铕(Eu)颗粒标记氯氰菊酯抗体,研发针对高效氯氰菊酯检测的时间分辨免疫荧光检测卡.在对虾样品中添加高效氯氰菊酯标准品分析检测卡的准确度、 精确度等指标时,研究发现氯氰菊酯检测卡的平均相对误差在4.24％ ～8.66％之间,检测卡的批内变异系数在3.0％ ～8.6％之间,批间变异系数在4.8％ ～6.6％之间,最低检测限为0.35 ng/mL,检测范围为1～19.3 ng/mL,四参数拟合曲线决定系数R2=0.9990;检测卡检测结果与气相色谱检测结果的相对误差在8％ ～20％之间,线性相关系数R=0.9899.结果表明本技术方法可以应用于水产品中高效氯氰菊酯的快速定量检测.

The enzymatic hydrolysis parameters for preparation of antioxidant peptide from seahorse protein were optimized.Base on the results of single factor experiment,content of free amino acids,reducing power and scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were chosen as the measure indexes,the influences of enzyme species,pH value,enzymolysis temperature,reaction time and enzyme in solid matter for enzymatic hydrolysis were inspected.Optimum hydrolysis conditions were confirmed as enzyme in solid matter,6000 U/g;enzymolysis temperature,45℃;and reaction time,9 hours.Content of the free amino acids,reducing power and scavenging activity on DPPH radical under optimum conditions was 4.198 mg/mL,92.758％ and 1.091,respectively.The optimum hydrolysis conditions were generated and laid a solid foundation for the development of health care products.

In this study, the structural changes of gut microbiota in response to abalone viscera protein hydrolysate (AVPH) were examined in alcohol induced injured mice. The gut microbiota of mice without treatment (blank), alcohol treatment (control), and alcohol+AVPH treatment (sample) were profiled with Illumina sequencing of the V4 region of 16S rDNA genes. The sequencing results revealed that 705, 654, and 888 of Operational Taxonomic Units (OTUs) were determined in the blank, control, and sample groups, respectively. Compared with the control and blank groups, the AVPH had a higher diversity index and a larger number of reads subjecting to Bacilli (class), Lactobacillales (order), Lactobacillaceae (family), and Lactobacillus (genus) levels. However, insignificant difference of beta-diversity or principal component among control, blank, and sample groups was found. This study demonstrates that the in vivo oxidation inhibition pathway in alcohol induced injured mice by AVPH was not via modulating the gut microbiota structure.

Fresh Crassostrea angulata ~om Fujian was used as a raw material to produce oyster hydrolysate by enzymolysis using mixed enzymes,isolation and purification with an ultrafiltmtion membrane,and freeze-drying.Amino acid composition of the oyster hydrolysate was analyzed using an automatic amino acid analyzer.The hypoglycemic effects of the oyster hydrolysate were evaluated using streptozotocin (STZ)-induced diabetic mice that were divided into seven groups:normal control group,model control group,metformin group (Met,at a daily dose of 0.5 g/kg BW),homogenate group (oyster homogenate,at a daily dose of 2.0 g/kg BW),low-dose group (enzymolysis product,at a daily dose of 0.5 g/kg BW),medium-dose group (enzymolysis product,at a daily dose of 1.0 g/kg BW),and high-dose group (enzymolysis product,at a daily dose of 2.0 g/kg BW).After 28 d of intragastric administration,the body weight,fasting blood glucose level,and glucose tolerance of mice were measured,and the mechanism of hypoglycemic activity of the oyster hydrolysate was explored.Nutritional component analysis showed that the oyster hydrolysate had high protein content and low fat content.It also showed that essential amino acids accounted for 42.34％ of the total amino acids and 73.43％ of the nonessential amino acids;this is in agreement with the ideal protein model suggested by the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO).Furthermore,the oyster hydrolysate had no effect on the blood glucose level in normal mice,but improved symptoms such as body weight loss,heavy drinking,and binge eating in diabetic mice.Oyster hydrolysate administered at low,medium,and high doses significantly reduced fasting blood glucose levels in diabetic mice by 21.09％,32.12％,and 37.05％,respectively (p＜0.05),and the glucose area under the curve (AUC) value by 51.67,47.30,and 43.41,respectively (p＜0.01).The results indicate that the oyster hydrolysate exerts potent hypoglycemic activity in diabetic mice.

采用福建省水产研究所研制的“河豚毒素半定量快速检测试剂盒”,在双斑东方鲀人工繁殖及苗种培育过程中,对不同繁殖群体亲种,不同繁殖群体交配所得受精卵、胚胎及仔稚幼鱼的河豚毒素进行跟踪检测.结果表明:不同繁殖群体亲鱼组织器官毒性差异大,亲鱼眼珠、精巢为阴性,皮肤、肌肉、肝脏、卵巢呈现不同阳性检出率,分别为5.4％、16.2％、40.5％、40.5％;养殖繁殖群体亲鱼皮肤、肝脏为阳性,野生繁殖群体亲鱼肝脏和雌性性腺河豚毒性阳性检出率比养殖繁殖群体亲鱼高;野生群体雌性亲鱼与野生群体雄性亲鱼、养殖群体雄性亲鱼交配组受精卵河豚毒素均为阳性,早期胚胎为阳性,后期胚胎为阴性;养殖群体雌性亲鱼与野生群体雄性亲鱼、养殖群体雄性亲鱼交配组受精卵、胚胎河豚毒素皆为阴性;野生群体雌性亲鱼与野生群体雄性亲鱼交配组仔稚鱼河豚毒素为阳性,早期幼鱼为阳性,后期幼鱼为阴性;野生群体雌性亲鱼与养殖群体雄性亲鱼交配组仔鱼河豚毒素为阴性,稚鱼、早期幼鱼为阳性,后期幼鱼为阴性;养殖群体雌性亲鱼与野生群体雄性亲鱼、养殖群体雄性亲鱼交配组仔稚鱼河豚毒素皆为阴性.

研究喷雾干燥制备牡蛎肽粉对体内抗氧化活性的影响.以新鲜葡萄牙牡蛎为原料,通过选择性复合酶解后喷雾干燥制备牡蛎肽粉,分别测定小鼠丙二醛(MDA)、蛋白质羰基、超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)含量等指标考察牡蛎肽粉对小鼠抗氧化活性的影响.结果显示,与模型对照组相比,经过喷雾干燥后的牡蛎肽粉不同剂量组均可极显著地降低小鼠血清丙二醛含量和蛋白质羰基含量(p＜0.01),并显著提高超氧化物歧化酶和还原型谷胱甘肽的含量(p＜0.01).结果表明喷雾干燥制备的牡蛎肽粉具有良好的体内抗氧化作用.

The changes in flavor volatile compounds in discus abalone Haliotis discus hannai Ino before and after cooked were analyzed by an electronic nose and solid phase micro-extraction ( SPME) coupled with gas chromatog-raphy-mass spectrometry ( GC-MS) to reveal the main factors causing flavor formation by cooking processing. The results showed that the change in volatile compounds generated by cooking processing was sensitively detected by the electronic nose. The principal component analysis indicated that there was significant difference in volatile com-pounds in discus abalone before and after it was cooked. GC-MS analysis demonstrated that the volatile compounds were changed significantly before and after cooking, and that 42 volatile compounds were detected in uncooked dis-cus abalone and 61 volatile compounds in cooked discus abalone, including ethanol, aldehydes, ketones, hydrocar-bons, esters, acids, aromatic series, and nitrogen containing compounds. In the flavor volatile compounds, benzal-dehyde (68. 15%) and 1-octen-3-ol (9. 22%) were of the major compounds in the raw discus abalone,while 2, 3-butanediol(22. 80%) and(2R,3R)-(-)-2,3-butanediol(19. 84%) were the major volatile compounds in the cooked discus abalone. It was concluded that the electronic nose and solid phase micro-extraction ( SPME) cou-pled with gas chromatography-mass spectrometry ( GC-MS ) can perfectly analyze the changes in flavor volatile compounds in discus abalone processed.