Antioxidant and Angiotensin-Converting Enzyme Inhibitory Activities of Protein Hydrolysates Prepared from Threadfin Bream (Nemipterus spp.) Surimi By-products

Solid wastes from threadfin bream (Nemipterus spp.) surimi production composed of head and frame were hydrolyzed by various commercial proteases (Alcalase, Flavourzyme, Neutrase, Protamex, papain, and pepsin) to produce protein hydrolysates with bioactive properties. An Alcalase-hydrolyzed sample at 24.4% degree of hydrolysis (DH) displayed the highest antioxidant activity based on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) assay, and potassium ferricyanide method. In addition, it showed an inhibitory activity toward angiotensin-converting enzyme (ACE) of 25.5%. Antioxidant activity of threadfin bream by-product hydrolysates increased with hydrolysis time and reached the highest DPPH activity after 6 h, while that hydrolyzed for 3 h showed the highest reducing power based on FRAP and potassium ferricyanide assays. In addition, ACE inhibitory activity was found to be at an optimum after 3 h of hydrolysis. The hydrolysates (1 mg/mL) also retarded oxidation of a linoleic acid emulsion system to a similar extent as 0.1 mg/mL 3-tert-butyl-4-hydroxy anisole (BHA), indicating a potential use in the food system. Protein hydrolysates from threadfin bream surimi by-products could be tailor-made to possess both antioxidant and ACE inhibitory activity through controlling DH of Alcalase-catalyzed reactions.     

Effect of Enzyme Type on the Antioxidant Activities and Functional Properties of Enzymatic Hydrolysates from Sea Cucumber (Cucumaria frondosa) Viscera

ABSTRACT

Effects of various proteases were obvious on the amino acid composition, antioxidant activities, and functional properties of enzymatic hydrolysates prepared from sea cucumber viscera (SCV). Alcalase, trypsin, and flavourzyme conferred better antioxidant activities on hydrolysates and endowed them with a higher degree of hydrolysis, coming to 19.08, 32.38, and 15.94%, respectively. For functional properties, neutrase- and flavourzyme-derived hydrolysates showed better foaming properties. Amino acid composition suggested that papain, neutrase, alcalase, and bromelain granted the hydrolysate’s high nutritional quality. The results indicated that SCV hydrolysates could be used in food systems to impart desirable characteristics to products.     

Kinetics of Antioxidant-Producing Maillard Reaction in the Mixture of Ribose and Sea Cucumber (Stichopus japonicus) Gut Hydrolysates

Maillard reaction products (MRPs) were produced in sea cucumber gut hydrolysates (SCGHs)-ribose system at 75°C, 85°C, and 95°C with a pH value of 7.0 or 8.0 for 12 hour. The parameters of amount of intermediate products, fluorescence intensity, browning development, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability showed a positive correlation with each other in SCGHs-ribose system. The amount of intermediate products, fluorescence intensity, and browning development followed the zero-order kinetic with activation energy from 62 to 91 kJ/mol in the Maillard reaction of SCGHs-ribose system. Moreover, 2-methyl-butanal has positive and significant correlation with DPPH radical scavenging ability of SCGHs-ribose MRPs. These results suggest that Maillard reaction improves the antioxidant activity of SCGHs, which contributes to the generation of intermediate, fluorescent, browning, and volatile products of SCGHs-ribose system.     

Assessing the Antioxidant Activity of the Ultrafiltration Fractions From Silver Carp Protein Hydrolysate by Different Antioxidant Methods

Silver carp protein hydrolysate (SH) was fractionated by 5- and 1-kDa cutoff ultrafiltration (UF) membrane, and three ultrafiltrates were obtained: SHI (>5 kDa), SHII (1–5 kDa), and SHIII (<1 kDa) which were screened for antioxidant activity by five in vitro assays. The SHIII showed higher 2, 2-azobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) values, inhibiting effects for liposome oxidation induced by Fe³⁺/V_C. The SHII showed higher ferric reducing power (FRAP) and inhibiting effects for liposome oxidation induced by 2, 2′-azobis (2-methylpropionamidine) dihydrochloride (AAPH). However, the SHI showed the highest 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging activity. Therefore, the antioxidant activity of silver carp protein hydrolysates was mainly associated with their molecular weight distribution, and the antioxidant assays used to test different samples gave comparable results.     

Evaluation of Processing Methods on the Nutritional Quality of Sea Cucumber (Apostichopus japonicus Selenka)

The close relationship between the consumption of sea cucumber and health status stems from the nutritional and functional substances found in sea cucumber that play a key role in the prevention of different diseases. The processing methods have a great effect upon the nutritional and functional substances in sea cucumber. A superior product should not only be abundant in nutritional substances, but also require an easy and cost effective processing method. This study compared the influence of processing methods on the stability of macronutrients and micronutrients present in sea cucumber in order to recommend products that could provide high nutritional quality. The five processes investigated were hot air drying (60°C), vacuum freeze-drying (0.5 cm thickness material, 5–10 Pa vacuum degree, ?85 to ?90°C cold trap temperature, ?25°C material temperature), sun drying, double-distilled water cooking followed by sun drying, and 3.5% sodium chloride solution cooking followed by sun drying. A significant decrease of all the chemical compositions determined was shown in all the sea cucumber products treated by the five processing methods (p < 0.05). Among all the processing methods, hot air drying resulted in the highest nutritional quality of the product. Moreover, hot air drying can be operated easily with low cost.     

附着刺参苗的行为观察

通过室内和海上实验,分别对体长１ｃｍ、３ｃｍ、５ｃｍ三种规格随着刺参苗的摄食、栖息和运动行为进行了观察研究。发现随着参苗有明显的昼伏夜出习性。白天刺参苗躲藏于随着基的背光处,日落后２ｈ左右参苗开始在附着基上爬行、摄食,可持续４～５ｈ,随着参苗继续躲藏于背光处。参苗夜间的摄食、运动导致了参苗从随着基上脱落,个体越大,运动能力越强,脱落比例越大。此外,自然海区的风浪作用也对参苗脱落起着重要作用。     

Optimization of Hydrolysis Conditions for the Production of the Angiotensin-I Converting Enzyme Inhibitory Peptides from Sea Cucumber Collagen Hydrolysates

The effects of different hydrolysis conditions on the angiotensin-I converting enzyme (ACE) inhibitory properties of giant red sea cucumber (Parastichopus californicus) collagen hydrolysates were investigated. Optimal conditions predicted by central composite rotatable design (CCDR) modeling for producing ACE inhibitory peptides were found to be 54.9°C, 1.76 h and an enzyme to substrate (E/S) ratio of 0.064. Compared with experiments performed under these optimal conditions, the predicted degree of hydrolysis (DH) and ACE inhibitory activities had error rates of 3 to 6%. Under optimum conditions, the molecular weights of collagen hydrolysates were less than 6.5 kDa. Lineweaver-Burk plots suggest that the collagen hydrolysates acted as competitive inhibitors with an inhibition constant (Ki) value of 0.706 mg/mL.     

Spawning and Larval Development of the Four‐Sided Sea Cucumber,Isostichopus badionotus (Selenka 1867), under Controlled Conditions

The objective of this study was to generate information for the development of Isostichopus badionotus culture protocols by determining optimal spawning methods, egg production, length and duration of larval stages, and larval habitat settlement preferences. Studies performed during the spawning seasons of 2010 and 2011 showed that this organism spawns between July and November without mechanical or chemical induction. The larval development has five well‐defined stages: early, mid and late auricularia, doliolaria, and pentactula. Juveniles (654.3 µm length) were obtained on average 25 days after fertilization (DAF), when larvae were incubated at 25 ± 1 C. No preference was observed for any of the materials used for settlement of the larvae. The easy adaptation of I. badionotus to the culture conditions and the competence of the larvae to complete metamorphosis while feeding solely on microalgae concentrates make this species an excellent candidate for farming. However, it is necessary to continue designing techniques for larval culture and to establish appropriate captivity conditions that allow multiannual reproduction.     

海参中单糖检测方法的建立及含量测定

本研究建立了高效阴离子交换–脉冲安培检测法(HPAEC-PAD)分析海参(Apostichopus japonicus)多糖的单糖组成及含量测定。通过比较3种蛋白去除法对海参多糖含量的影响,优化海参多糖纯化方法;比较淋洗液浓度对单糖分离效果的影响,优化海参中单糖分离的色谱条件。结果显示,Sevag法去蛋白的效果最好,在淋洗液为20 mmol/L Na OH的条件下分离岩藻糖、氨基半乳糖、氨基葡萄糖、半乳糖、甘露糖、阿拉伯糖、葡萄糖和乳糖;在淋洗液为160 mmol/L Na OH和200 mmol/L Na AC的条件下分离葡萄糖醛酸和半乳糖醛酸。各单糖标准曲线的线性相关系数不低于0.998,检出限为2.5~50μg/L,加标回收率在83.6%~113.1%之间。该方法可测定海参多糖的中性糖、氨基糖和糖醛酸,可作为海参及其制品中的单糖分析及定量方法,为相关标准的制定提供参考。     

Evaluation of the growth and survival rate of the Caribbean Sea cucumber,Isostichopus badionotus (Selenka, 1867), early juveniles produced in captivity

Isostichopus badionotus is a new commercially important sea cucumber in the Caribbean that has rapidly shown signs of overexploitation. There is no information on the growth and survival rates for wild or hatchery‐produced early juveniles. Hatchery production and determination of juvenile growth rates are essential for future aquaculture efforts for this species. In this study, daily growth rate (GR), specific growth rate (SGR), weight gain (WG), survival rate (SR), and weight–length relation (WLR) were evaluated using small (0.01 g) and large (0.25 g) hatchery‐produced early juveniles under laboratory conditions. Growth was rapid compared to other cultured sea cucumber species; maximal values of SGR and WG were obtained for small juveniles (4.58%/day and 2,191%, respectively), while maximal GR and SR were obtained for large juveniles (0.29 mm/day and 99.17%, respectively). A wide range of sizes was evident at the end of each trial. The WLR of small and large juveniles fit with a negative allometric growth trend. This study highlights the potential production of I. badionotus as early juvenile stages withstand hatchery handling well and exhibit a high SR, reaching commonly released mean sizes (>0.7 g and 25 mm) after almost 3 months (120 days postfertilization).     

Protein Hydrolysates and Ultrafiltration Fractions Obtained from Krill (Euphausia superba): Nutritional,Functional, Antioxidant,and ACE-Inhibitory Characterization

ABSTRACT

Krill (Euphausia superba) was hydrolyzed by proteolytic enzymes in order to produce multifunctional bioactive peptides, and their functional properties were evaluated. Krill protein hydrolysate (KPH) by pepsin with 4-h hydrolysis showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and angiotensin I converting enzyme (ACE) inhibitory activities. The solubility and foaming properties of KPH were higher than those of the unhydrolyzed krill protein at a wide range of pHs. KPH was further fractionated based on molecular weight. The 1- to 3-kDa peptide fraction exhibited the highest DPPH scavenging activity (IC₅₀ value of 0.5 mg/mL), oxygen radical absorbance capacity (497.39 ± 4.31 µM TE/mg fraction), 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid cation radical scavenging activity (48.41 ± 0.23 µM TE/mg fraction), and reducing power (110.40 ± 2.07 µM TE/mg fraction). However, the < 1-kDa peptide fraction exhibited a higher ACE inhibitory activity than that of other fractions. The 1- to 3- and < 1-kDa peptide fractions are rich in aromatic and hydrophobic amino acids, respectively.     

海参多肽的制备工艺优化及其抗氧化测定

本文对海参酶解工艺条件进行优化,提取液经超滤和冷冻干燥获得海参多肽,并测定其组成及抗氧化活性。结果显示,中性蛋白酶对海参最佳的酶解工艺为：加酶量1800U／g,酶解温度为55℃,酶解pH值为7．5,酶解时间为3h。经超滤与冷冻干燥后,多肽得率为6．9％,蛋白质含量达94．9％。且在抗氧化方面具有较好的清除羟自由基和超氧自由基能力。     

Extraction and Characterization of Pepsin Soluble Collagen from the Body Wall of Sea Cucumber Acaudina leucoprocta

Sea cucumber Acaudina leucoprocta is a potential alternative collagen source. However, the high levels of heavy metals contained in the body wall restricts its utilization. In this work, an efficient method was established to remove the heavy metals accumulated in the body wall of A.leucoprocta by demineralizing with 0.2 M ethylenediaminetetraacetic acid (EDTA). The resulting body wall of A.leucoprocta was then used for extracting pepsin-soluble collagen (PSC) with pepsin proteolysis. The PSC from the body wall of A.leucoprocta was obtained with a yield of 43.99 ± 0.65% (dry weight) and high purity. Maximum and minimum solubility for the isolated PSC in 0.5 M acetic acid was observed at pH 2.66 and 4.43, respectively. The solubility was remarkably decreased in the presence of NaCl. The denaturation temperature of PSC rehydrated in 0.5 M acetic acid was measured as 25.4°C. The PSC was characterized as type I collagen, which consists of three α₁ chains without α₂ chain. Interestingly, α₁ chain in PSC showed two isoforms with the pI values of 4.02 and 4.29. The heavy metals existing in PSC were all below the contaminant limit of edible gelatin. The PSC isolated from the body wall could be an alternative to mammalian collagens.     

养殖仿刺参溃烂病病因初探

从患病仿刺参分离获得的两株优势菌2004—A1和2004—A2,经人工感染回接健康参试验,均获得与原发病相同症状,证实此两株菌是病原菌。另通过人工创伤感染试验确认海参受外伤后,容易感染发生溃烂病。     

氨氮胁迫对不同规格刺参(Apostichopus japonicus)存活及非特异性免疫酶活性的影响

本实验以小[(6.05±0.44)g/头]、中[(14.68±1.76)g/头]、大[(25.64±3.27)g/头]的3种不同规格刺参(Apostichopus japonicus)为研究对象,研究了氨氮胁迫浓度为0、2、4、6、8、l0 mg/L时对刺参体腔液中溶菌酶(LSZ)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)活性的影响.结果显示,在氨氮浓度为8 mg/L时,第10天后,3种不同规格刺参均出现吐肠、化皮,直至死亡.氨氮浓度为10 mg/L时中规格刺参存活率最低,为86.7％.在氨氮浓度为2 mg/L和4 mg/L时,小、中、大规格刺参的3种非特异性免疫酶活性在第4天显著升高,与对照组(氨氮浓度低于0.05 mg/L)差异显著(P＜0.05);第7、10、13天时,LSZ、CAT、SOD酶活性与第4天相比,差异不显著(P＞0.05).在氨氮浓度为6、8、10 mg/L时,LSZ、CAT、SOD酶活性在第7天达到最高,与对照组差异性显著(P＜0.05).氨氮胁迫对3种不同规格刺参的非特异性免疫酶活性的影响存在差异,在同一氨氮浓度胁迫下,敏感性依次为中规格＞大规格＞小规格.研究表明,氨氮胁迫会对刺参存活与免疫产生一定影响,在刺参养殖过程中,水环境中氨氮浓度最好控制在6 mg/L以内.     

利用塑料大棚晒水进行刺参种苗越冬试验

2002年11月~2003年4月,在瓦房店市交流岛前哨海参育苗厂800 m3水体室内水泥池中,投放平均规格为5000头/kg,刺参种苗196 kg(98万头)。在没有锅炉进行水加温条件下,利用600 m3水体塑料大棚晒水进行刺参种苗越冬,水温2.0~13.2℃,平均盐度31.2,pH值8.2,DO>7 mg/L,经过近6个月的培育,共培育出规格为876头/kg的参苗836 kg(73.2万头),成活率74.7%,总重量增加了8.5倍。     

不同浒苔型饲料对幼刺参(Apostichopus japonicus)生长、消化及非特异性免疫的影响

以浒苔、石莼、豆粕、扇贝边、葡萄糖、贝壳粉、维生素和矿物质预混料为原料配制刺参(Apostichopus japonicus)饲料,利用酿酒酵母菌菌液和碱性蛋白酶制剂对刺参饲料进行4种不同的处理,得到5组实验用饲料,分别为对照组、发酵组、酶解组、复合组和鲜浒苔组.将上述饲料饲喂初始体重为(1.92-0.02)g的幼刺参42 d,每种饲料设3个重复,每个重复30头幼刺参.结果显示,不同的浒苔型饲料对幼刺参的存活率(SR)、增重率(WGR)、特定生长率(SGR)和饲料系数(FC)有显著影响(P＜0.05),而对其脏壁比(R)无显著性影响(P＞0.05).复合组和鲜浒苔组SR要显著高于酶解组(P＜0.05),而与其他2组均无显著性差异(P＞0.05).鲜浒苔组WGR远高于其他各组(P＜0.05),而复合组和发酵组WGR显著高于酶解组(P＜0.05),与对照组差异不显著(P＞0.05).幼刺参的SGR规律与WGR一致.鲜浒苔组FC显著低于对照组、发酵组和酶解组(P＜0.05),与复合组差异不显著(P＞0.05).随着摄食饲料时间的推移,对照组、发酵组、复合组和鲜浒苔组淀粉酶(AMS)活力先升高再下降后趋于稳定,而酶解组一直呈现下降趋势.酶解组纤维素酶(Cellulase)活力呈现一直下降的趋势,而其他组呈现波动变化,且均高于初始活力值.随着摄食饲料时间的推移,除酶解组外,其余各组胰蛋白酶(TRY)活力前后时间点变化差异不大,且每个采样点幼刺参TRY活力大小顺序始终是对照组＞复合组＞鲜浒苔组＞发酵组＞酶解组.不同浒苔型饲料饲喂的幼刺参体腔液中酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和超氧化物歧化酶(SOD)活力均有显著性差异(P＜0.05).鲜浒苔组ACP活力最大,且与复合组无显著性差异(P＞0.05),而显著高于其他3组(P＜0.05).鲜浒苔组和复合组AKP活力显著高于酶解组和对照组(P＜0.05),与发酵组无显著性差异(P＞0.05).复合组SOD活力最大,且显著高于发酵组和酶解组(P＜0.05),而与对照组和鲜浒苔组均无显著性差异(P＞0.05).由此得出,幼刺参在摄食先酶解后发酵的饲料后能够得到良好的生长效果,并可改善自身肠道消化,维持正常免疫.这为解决刺参饲料原料短缺以及浒苔高值化利用提供了依据和方法.     

3种水质调控方式下参池沉积物酶活性的比较研究

2015年10月至2016年9月对自然纳潮、微孔曝气、养水机3种水质调控方式下海参池塘沉积物中淀粉酶、蛋白酶、碱性磷酸酶、脱氢酶的活性进行了跟踪比较研究。研究结果显示,3种水质调控方式下池塘沉积物中的淀粉酶活性年变化为0.126~0.880 mg/g,年均值(0.410±0.180) mg/g,蛋白酶活性年变化为0.024~0.472 mg/g,年均值(0.190±0.103) mg/g,碱性磷酸酶活性年变化为0.068~1.042 mg/g,年均值(0.340±0.196) mg/g,脱氢酶活性年变化为12.092~52.794 mL/g,年均值(26.980±8.295) mL/g。3种水质调控方式下池塘沉积物中蛋白酶、碱性磷酸酶及脱氢酶活性均在自然纳潮池塘中均值最高,变化幅度最大;淀粉酶活性均值则在自然纳潮池塘中最低,养水机池塘最高,这与养水机池塘有机质最低,自然纳潮池塘最高,养水机池塘沉积物的细菌多样性最高,真菌数量最多有关。表明养水机能够快速去除沉积物中氮、磷有机化合物,有利于池塘的正常物质循环。本研究从沉积物酶活性的角度,探讨了养水机的作用效果与其他两种水质调控方式产生差异的机理。     

四种药物对刺参幼参毒性的初步研究

采用急性实验的常规方法—死亡概率法,用甲醛及几种抗生素对刺参幼参的急性毒性试验,结果表明:甲醛、氯霉素、环丙沙星、盐酸土霉素对刺参幼参24h半致死浓度(LC50)分别为:15.4×10-6、146.9×10-6、173.6×10-6、263.4×10-6;48h半致死浓度(LC50)分别为:13.6×10-6、107×10-6、144.4××10-6、244.7×10-6;安全浓度(SC)分别为:3.18×10-6、17.03×10-6、29.97×10-6、63.35×10-6。