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为了探究养殖花鳗鲡(Anguilla marmorata)鱼皮胶原蛋白的特性,利用胃蛋白酶结合酸抽提法提取花鳗鲡鱼皮胶原蛋白,SDS-PAGE电泳法展示胶原蛋白的电泳图谱,氨基酸分析仪测定胶原蛋白的氨基酸组成,分析胶原蛋白分子的紫外和红外光谱特征,用浊度实验和流变黏弹性实验探究胶原蛋白分子的体外自组装动力学特性。结果显示,花鳗鲡鱼皮胶原蛋白属于Ⅰ型胶原蛋白,氨基酸组成中甘氨酸、脯氨酸与羟脯氨酸三者的比值为8∶3∶2,胶原蛋白分子的最大紫外吸收峰出现在222.6 nm处,热变性温度28℃,红外光谱图和扫描电镜扫描结果图均表明经胃蛋白酶及酸溶得到的胶原蛋白保持了胶原原有的结构。胶原蛋白分子的体外自组装动力学曲线是含有迟滞期、成长期和稳定期的三阶段曲线。以上结果表明花鳗鲡鱼皮胶原具有接近淡水鱼皮胶原的特征。 相似文献
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以暗纹东方鲀(Takifugu obscurus)精巢组织为原料,采用酸提法提取鱼精蛋白。以得率为指标,通过正交实验,确定了最佳的提取参数。结果显示,提取鱼精蛋白的影响因素重要性依次为：提取次数>硫酸用量>硫酸浓度>95%乙醇用量;最佳提取工艺条件：硫酸浓度为0.2 mol/L、硫酸用量为2.5倍、提取次数为2次、95%乙醇用量为2.5倍。在此工艺条件下,暗纹东方鲀鱼精蛋白的得率为3.82%,蛋白含量达89.01%。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(Tricine-SDS-PAGE)可知,提取的粗鱼精蛋白有2个条带,分子量分别在25和20 kDa附近。分析其氨酸酸组成发现,暗纹东方鲀鱼精蛋白属于双鱼精蛋白,其中,精氨酸和丙氨酸含量相对较高,分别占31.40%和17.39%。本研究对暗纹东方鲀鱼精蛋白更好地应用在食品和医药领域具有重要意义。 相似文献
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暗纹东方鲀养殖关键技术浅析 总被引：1，自引：0，他引：1
<正>暗纹东方鲀俗称"河豚",其肉质鲜美,但含剧烈神经性麻痹毒素。暗纹东方鲀属海江生殖洄游鱼类,是我国长江下游地区重要的渔业资源。随着环境变化以及人类活动的影响,野生的暗纹东方鲀资源日渐稀少,江苏省扬中市水产技术研究所从1995年起开展了暗纹东方鲀的人工繁殖和养殖技术研究。目前,人工养殖的暗纹东方鲀已经全面取代了野生的暗纹东方鲀成为了食用鱼源。本文所要探讨的便是人工养殖暗纹东方鲀的一些关键技术。 相似文献
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为研究不同温度条件下盐度对暗纹东方鲀幼鱼越冬成活率的影响,于2012年底进行了盐度梯度试验,测定在不同温度(8、10、12、15℃)下,(50±2.32)g的暗纹东方鲀在不同盐度(0、1.5、3.0、4.5、6.0)下的越冬成活率,并通过大棚自然越冬试验进行验证。结果表明:同一温度条件下,在试验盐度范围内,随着盐度的升高,暗纹东方鲀的越冬成活率也逐渐提高;而在同一盐度下,随着温度的升高,暗纹东方鲀的越冬成活率也逐渐提高。当盐度为0时,暗纹东方鲀无法在自然温度下安全越冬;当盐度为4.5时,水温在8℃以上,暗纹东方鲀即能安全过冬,其越冬成活率达到80%以上。暗纹东方鲀大棚自然越冬试验证明,随着温度的下降逐渐增加水体盐度的方法经济而有效,当温度为15、12、10、9℃时,将相应的水体盐度调节为1.5、3.0、4.5、4.5,经过150 d的越冬期,两种规格(50~60 g和20~40 g)暗纹东方鲀的成活率分别为93.7%和80.5%。 相似文献
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为探究暗纹东方鲀(Takifugu fasciatus)TGF-β1基因与其生长性状的关联及作用,采用RACE技术克隆了暗纹东方鲀TGF-β1基因,分析了其基因结构与表达特征,并通过直接PCR扩增法对TGF-β1基因进行多片段比对,在此基础上筛选出与生长性状相关的SNP位点。研究发现TGF-β1基因cDNA全长共2 217 bp,包含5′UTR 51 bp, 3′UTR 1 014 bp, ORF 1 152 bp,编码383个氨基酸。氨基酸序列比对分析表明,暗纹东方鲀TGF-β1基因与红鳍东方鲀(Takifugu rubripes)同源性最高(92.82%)。进化树分析表明,暗纹东方鲀TGF-β1基因编码氨基酸序列与红鳍东方鲀聚为一支,与鲈形目鱼类亲缘关系最近(68.46%～73.33%)。qRT-PCR检测显示,TGF-β1基因在暗纹东方鲀8种不同组织(心脏、肝脏、脑、脾脏、肾脏、肌肉、中肠和鳃)都有表达,其中肾脏表达量最高,心脏和脾脏表达量次之。通过SNP位点不同基因型与个体生长性状的相关性分析,发现TGF-β1上的SNP g. 3908 C>A位点与暗纹东方鲀多项生长性状... 相似文献
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比较投饵率水平分别为3%、4%、5%、6%时,对暗纹东方鲀幼鱼生长、饲料效率的影响,以确定暗纹东方鲀幼鱼阶段的适宜投饵率。在23℃下,对规格为3.45～3.56g的暗纹东方鲀幼鱼进行30d的生长实验,结果表明:不同投饵率对暗纹东方鲀幼鱼的特定生长率、质量相对增加率、饲料效率、存活率均有显著影响(p<0.05);暗纹东方鲀幼鱼的适宜投饵率为4%,在投饵率为4℅时,质量相对增加率为116.04±0.04%,饲料效率达83.70±0.02%,特定生长率为111.49±0.03%/d,存活率为84.44±0.02%。 相似文献
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为研究盐度对暗纹东方鲀幼鱼生长和摄食的影响,将暗纹东方鲀幼鱼在盐度分别为0、10、17、26、40的水体中饲养6周,比较不同盐度下幼鱼存活率、特定生长率、摄食率和饲料系数的差异。结果表明:当盐度为40时,暗纹东方鲀幼鱼的死亡率为100 %;各盐度组的特定生长率无显著差异;幼鱼的摄食率随着盐度的增高而下降。在淡水环境中,暗纹东方鲀幼鱼的摄食率最高,达5.02 %;在高盐度水体中最低,为3.75 %。暗纹东方鲀幼鱼的饲料系数随着盐度的升高逐渐升高,淡水组和26盐度组与10、17盐度组有显著差异。 相似文献
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性腺型芳香化酶基因是一种调节雌雄激素平衡的基因。为初步探究暗纹东方鲀性腺型芳香化酶基因在性腺发育和性别分化过程中的作用,通过RACE及荧光定量PCR技术,成功克隆出暗纹东方鲀性腺型芳香化酶基因的cDNA全长序列,共1786 bp,编码514个氨基酸,与红鳍东方鲀及星点东方鲀同源性最高。氨基酸序列分析显示,该基因编码的蛋白是一种稳定的亲水性蛋白,但不存在信号肽序列;性腺型芳香化酶含有38个磷酸化位点、4个N-糖基化位点、2个跨膜保守结构域;其氨基酸序列在哺乳动物和鱼类中均十分保守,且含有跨膜区、Ⅰ-螺旋区、Ozol′s肽区、芳香化酶特异性保守区和亚铁血红素结合区等功能保守区。表达分析显示,暗纹东方鲀性腺型芳香化酶基因主要在肌肉和卵巢中表达,其次在其他组织中也有少量表达,而且在幼鱼不同发育期卵巢中的表达量呈现出逐渐升高再降低的表达趋势,在精巢中则基本不表达。研究结果表明,性腺型芳香化酶基因很可能参与了暗纹东方鲀雌鱼性腺分化后卵巢的发育、雌性特征的维持和其他组织的发育等过程。 相似文献
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Pornpimol Sungperm Chompoonuch Khongla 《Journal Of Aquatic Food Product Technology》2020,29(7):650-660
ABSTRACT The objectives of this study were to characterize physicochemical properties of two collagens, tilapia skin (TS) and hybrid catfish skin (HS). Angiotensin-converting enzyme (ACE) inhibitory peptides from extracted TS and HS collagen using pepsin were also determined. HS collagen had a higher amount of imino acid than TS collagen, while TS contained higher amounts of tyrosine (Tyr) and lysine (Lys). Fourier-transform infrared spectra of both collagens showed predominant helix structure. The HS collagen hydrolysate prepared by pepsin was fractionated using sequential ultrafiltration membranes, and the fraction with molecular weight (MW) <5 kDa showed the highest ACE inhibitory activity (p < .05). After cation exchange and two steps size exclusion chromatography, peptides showed ACE inhibitory activity of 72.06%. This study revealed that ACE inhibitory peptides derived from HS collagen could be developed as a functional food with potential antihypertensive properties. 相似文献
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草鱼鱼鳞胶原蛋白的提取及其部分生物学性能 总被引：5，自引：2，他引：3
以草鱼鱼鳞为原料, 分别提取鱼鳞中的酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(PSC), 着重开展了其包括热稳定性、体外酶降解性以及胶原海绵材料特性在内的相关研究, 并与哺乳动物来源的猪皮胶原(PC)相比较。实验结果表明, 制备所得的3种胶原蛋白均为典型的Ⅰ型胶原并具有完整的三螺旋结构; PC的热变性温度(41.6 ℃)明显高于ASC(34.8 ℃)和PSC(35.2 ℃); 3种胶原蛋白的体外酶降解性能受水解酶的种类、胶原蛋白提取方法、胶原蛋白来源、胶原蛋白受热历史以及蛋白的自组装程度影响。胶原蛋白酶、胰蛋白酶和木瓜蛋白酶对淡水鱼胶原均具有不同程度的降解能力, 但胶原蛋白酶的降解能力最强; 相同条件下, 3种胶原蛋白体外酶降解率依次为ASC>PSC>PC; 经热变性处理后胶原蛋白的体外酶降解率明显提高而经体外自组装处理后其体外酶降解率均出现不同程度的降低; 3种胶原样品冻干后得到的胶原海绵材料具有不同的机械性能和组织结构, ASC和PSC海绵是一种多孔但拉伸承受力较弱的海绵材料, 而PC则与之相反。 相似文献
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Shoshi Mizuta Mayu Nishizawa Fuuta Sekiguchi Kazuhiko Matsuo Yoshihiro Yokoyama Reiji Yoshinaka 《Fisheries Science》2013,79(5):841-848
Enzymatic solubilization of collagen from the skin tissue of diamond squid Thysanoteuthis rhombus, an underutilized resource in Japan, was attempted using an acid protease from the fungus Rhizopus niveus. This novel approach was compared with the conventional method using porcine pepsin. Both proteases were able to solubilize most of the skin collagen (>90 % of the total collagen) by performing the treatment in 0.5 M acetic acid at 4 °C for 72 h and at an enzyme/substrate ratio (w/w) of 1/10. The SDS-PAGE patterns of the solubilized collagen preparations were quite similar to each other, and two types of collagen (major and minor collagens) were purified from each preparation by cation-exchange column chromatography. These collagen types from the porcine pepsin-solubilized collagen showed similar features to those from the Rhizopus acid protease-solubilized collagen. These results suggest that the Rhizopus acid protease, a protease of non-animal origin, is applicable for solubilizing collagen in the skin of diamond squid. 相似文献
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Haibo Wang Yanping Liang Haiyin Wang Hanjun Zhang Min Wang Liangzhong Liu 《Journal Of Aquatic Food Product Technology》2013,22(3):264-277
In order to study the similarities and differences of physical-chemical properties of collagens from different parts of the fish body, acid-soluble collagens and pepsin-soluble collagens were isolated from skin, scale, and bone of grass carp (Ctenopharyngodon idellus), with yields ranging from 0.7 to 25.5% on the basis of dry weight. The content of acid-soluble collagen in skin and scale was much higher than that in bone according to the data of extraction yield. Collagens from grass carp have similar amino acid composition, protein patterns, and sequences of primary structure. Thermal transition temperatures of collagens from different parts of grass carp body decreased in the order of bone > skin > scale. The pepsin-soluble collagens showed more resistance to collagenase, compared to acid-soluble collagens extracted from the same part of the grass carp body. All grass carp collagens did not induce a significant cytotoxic effect according to the results of in vitro cytotoxicity test. 相似文献
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Min Zhang 《Journal Of Aquatic Food Product Technology》2013,22(1):44-58
Fish skin collagen solutions (0.3%, w/v) were treated using N-hydroxysuccinimide activated adipic acid (NHS-AA) as a cross-linking agent. Partial important physicochemical properties such as the molecular weight, the morphologies, the interaction between molecules, and especially the thermal stability of the cross-linked collagens were examined by various methods. Results from sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated the formation of cross-links, and thus the molecular weight of collagen was increased. Atomic force microscopy (AFM) suggested that the fiber diameter of cross-linked collagens was larger than that of collagen control. Scanning electron microscopy (SEM) showed that the number of pores from collagen sponges was reduced, accompanied by the rise in their pore sizes. According to Fourier transform infrared spectroscopy (FTIR) spectra, the value of amide III/amide I increased as a small amount of NHS-AA was used. Then it decreased and remained at a relatively constant level as NHS-AA was further added, which might indicate the altered interaction between collagen molecules due to cross-linking. Most important is the improved thermal stability of the cross-linked collagens as determined by differential scanning calorimetry (DSC). The reason for this improvement was further exposed by two-dimensional (2D) FTIR with temperature as perturbation. 相似文献
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Jun Cao Qingfei Duan Xiaoyan Liu Xuanri Shen 《Journal Of Aquatic Food Product Technology》2013,22(8):837-847
Pepsin-soluble collagen (PSC) was extracted and purified from wasted skin and bone of the golden pompano by acetic acid-pepsin method. The result showed that the PSCs extraction yields of skin and bone were 21.81% and 1.25% (wet weight), 62.21% and 1.78% (on the basis of lyophilized dry weight), respectively. Golden pompano skin and bone PSCs contained the typical chain of α and β dimers, and they were preliminarily judged to belong to type I collagen. The skin PSC had similar amino acid composition to bone PSC, which is rich in glycine, alanine, proline, and hydroxyproline. After addressing the pepsin, three helical structure of PSCs were intact, and their natural structures largely remained. The denaturation temperatures of skin and bone PSCs were 37.04°C and 38.23°C, respectively. Solubility results showed that the skin and bone PSCs solubility was the largest at pH = 3. The solubility of skin and bone PSCs was stable at NaCl concentrations lower than 3%. In addition, two PSCs in acid and low salt conditions had good solubility. This study demonstrated that golden pompano skin and bone could be used as good materials to extract PSC, representing an economic benefit and added value.Abbreviations: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis; FTIR: Fourier transform infrared; DSC: Differential scanning calorimetry; XRD: X-ray diffraction 相似文献
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Wanticha Savedboworn Soottawat Benjakul Sittichoke Sinthusamran Hideki Kishimura 《Journal Of Aquatic Food Product Technology》2017,26(3):248-257
Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from rohu skin with the yield of 64.2 and 6.8% (dry weight basis), respectively. Both collagens had glycine as the major amino acid with imino acid content of 196–202 residues/1,000 residues and were characterized as type I collagen with molecular composition of (α1)2α2-heterotrimer. Fourier transform infrared spectra of both collagens were similar, with no shift in wavenumber of all amide bands. The Tmax value of ASC and PSC was 36.40 and 35.48°C, respectively. The zero surface net charge of ASC and PSC was found at pH 5.9 and 5.3, respectively. 相似文献
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分别用热酸抽提法、碱抽提法和胃蛋白酶消化法分离豚链球菌NUF849的类M蛋白，通过SDS-PAGE和Western-blotting印迹法分析比较三种方法所提类M蛋白的组成和抗原性差异。结果表明，热酸抽提法分离制备的类M蛋白得率和纯度都较其它两种方法高，蛋白的抗原性保持较好。采用热酸提取法提取5株豚链球菌NUF633、NUF693、 NUF701、 NUF812、 NUF849和2株停乳链球菌SD、L2的类M蛋白，其SDS-PAGE图谱明显不同，种间差异明显, 豚链球菌主要类M蛋白的分子量分别为60、48、37、30、27、24和15kDa，停乳链球菌主要类M蛋白的分子量分别为68、48、42、37、29、26、23、21、19、17、15、14和11kDa。Western-blotting结果显示，兔抗豚链球菌血清可与豚链球菌的两条类M蛋白带结合，分子量分别为60和37kDa；与停乳链球菌的两条类M蛋白带结合，分子量为68和37 kDa。两种链球菌主要类M蛋白组成及其抗原性差异明显。 相似文献
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This study investigates an efficient method to extract collagen from the skins of sea bass. The ultrasonic treatment (20 kHz with amplitude of 20–80 %) was applied to fish skin for 24 h after the addition of 0.01, 0.05, and 0.1 M acetic acid (1:200 sample/acetic acid, w/v). As a result, the rate (Ki) of the collagen yield increased depending on the amount of acid added, the duration of ultrasonic treatment and the amplitude of the ultrasonic waves. The subunit compositions of the extracted components were analyzed by SDS-PAGE, and the main components were determined as collagen, including the α1 (α3), α2, β, and γ chains. And in addition to collagen, some unknown components were also observed after a longer period of ultrasonic treatment. Therefore, we had to optimize the efficient extraction conditions for pure collagen while minimizing the creation of unknown components. The most effective extraction condition for collagen by ultrasonic treatment was 80 % amplitude with 0.1 M acetic acid for 3 h of treatment. It was found that the component extracted by the ultrasonic treatment was indeed collagen, since there were no changes in the main components of collagen after pepsin treatment. 相似文献
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Kim Hyun Kyung Kim Young Ho Kim Yun Ji Park Hyun Jin Lee Nam Hyouck 《Fisheries Science》2012,78(2):485-490
In this study we investigated the effects of ultrasonic wave treatment on the extraction yield of acid-soluble collagen from
sea bass skins. Two extraction methods were compared: a 24 h acid treatment using 0.5 M acetic acid (1:200 sample/acid, w/v)
and an extraction using ultrasonic treatment after the addition of a 0.5 M acetic acid solution. The results indicated that
the extraction yield of collagen increased with the ultrasonic treatment, with the extraction rate increasing rapidly at higher
amplitudes of ultrasonic treatment. The subunit compositions of the collagen extracted by ultrasonic treatment were analyzed
by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which revealed that the α1(α3), α2, and β chains of collagen
were present early in the ultrasonic treatment. An unknown component, believed to be a product of collagen degradation induced
by the ultrasonic treatment, was detected only after a longer treatment time. The component extracted by the ultrasonic treatment
was determined to be collagen based on the finding that there were no changes in the main components of collagen, specifically,
the α1(α3), α2, and β chains, following pepsin treatment. 相似文献