鱼精蛋白的提取纯化及应用研究进展

鱼精蛋白是一种存在于各种鱼类精巢组织中的多聚阳离子肽,富含精氨酸,呈碱性,具有促进细胞繁殖发育、增强肝功能、抑制肿瘤生长繁殖等功能,在医学和保健领域有着广泛的应用;此外还具有显著的抗菌活性,可以开发成一种天然防腐剂.本文从分析鱼精蛋白的基本特性入手,重点介绍了鱼精蛋白的提取和纯化工艺,并讨论了其在食品、医学上的应用及分...     

鱿鱼鱼精蛋白的提取工艺优化研究

通过单因素和正交试验研究了鱿鱼精巢提取鱿鱼鱼精蛋白的优化工艺,并对优化工艺提取的鱿鱼鱼精蛋白进行了分析.结果表明,最佳提取工艺为硫酸浓度0.3M、硫酸用量为3倍、提取次数2次、冷乙醇用量为3倍;该条件下,鱿鱼鱼精蛋白的得率为3.42％,蛋白质含量达93％,且氨基酸组成齐全,碱性氨基酸占26.20％,其中精氨酸占9.61％,Tricine-SDS-PAGE电泳结果分析表明其组成成分比较复杂.     

特别推荐

<正>水霉净本品具有强氧化作用,可以破坏或分解真菌的细胞壁中的甘露聚糖和甘露聚糖-蛋白质复合物,迅速扩散进入细胞内,氧化细胞内的酶或RNA、DNA,从而致死菌原体。此外,本品还可以同细菌的细胞壁中的脂蛋白或细胞膜中的磷脂质、蛋白质发生化学反应,从而使细菌的细胞壁和细胞受到破坏(即所谓的溶菌作用),细胞膜的通透性增加,细胞内物质外流,使其失去活性。     

鱼精蛋白的特性及开发利用

鱼精蛋白是存在于鱼精巢中的一种碱性蛋白质,是一种高效、安全、值得开发的新型天然食品防腐剂.本文综述了鱼精蛋白的性质、提取、纯化、抑菌机理及其应用.     

暗纹东方鲀鱼精蛋白的提取工艺优化研究

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。     

猪喘气病的发病特点及防控措施

<正>猪肺炎支原体是一种介于病毒与细菌之间的特殊病原,其结构组成与细菌类似,但缺少细胞壁,多种针对细胞壁的抗生素对其效果甚微,同时由于缺少细胞壁,病原形状不规则。肺炎     

嗜水气单胞菌拮抗菌的筛选

用平板划线法或点种法对212株分离自养殖水体和水产动物胃肠道的细菌进行筛选,得到1株对嗜水气单胞菌(Aerom onas hydrophila)有拮抗作用的细菌MLS-7。经过生理生化的初步鉴定,该菌为粪肠球菌(Entero-coccus faecalis)。MLS-7上清液抑菌试验和共培养试验表明其对嗜水气单胞菌有较强的抑菌作用。进一步对MLS-7抑菌谱进行测定,确认其对鱼源的病原菌有一定的抑菌作用。     

抗菌素在鱼病防治中的应用

性状与作用：土霉素为黄色晶体，无臭、味略苦，性稳定，易溶于水。其主要作用是抑菌，高浓度时有杀菌作用。土霉素对鱼类的最低抑菌浓度为0．31～10μg／ml，多数为0．31～1．25μg／ml。治病范围广，毒性小，细菌耐药力产生很慢。肠道对其吸收很快，排出也快。     

具抑菌活性海洋微生物的筛选

从采集于上海郊区海域的海水和海泥样品中分离出 193株海洋细菌，从中筛选抑菌活性菌株。 分别利用琼脂块法和管碟法对分离出的菌株进行初筛和复筛，结果显示，有 29株海洋细菌具有抑菌活性， 占总分离菌株的 15%，其中抑菌能力较强的 G4，抑菌谱较广，能同时抑制革兰氏阳性和阴性指示菌。     

应用山莨菪碱与呋喃唑酮治疗淡水白鲳白皮病一例

据目前资料可知由细菌引起的鱼病多为革兰氏阴性细菌所致,因革兰氏阴性细菌能分泌内毒素内毒素是革兰氏阴性细菌细胞壁外膜的构成成分,其主要成份脂多糖是一种高分子,因其化学结构独特,且具有广泛的生物活性,它能对肌体产生不利的反应(致死,出血坏死,发热,抑制酶系统等)。当用药物杀死革兰氏阴性细菌时,由于细菌解体,反而使     

地中海弧菌Vibrio mediterranei 117-T6的溶藻活性

很多藻类致病菌具有广谱溶藻活性。本研究发现紫菜黄斑病致病菌地中海弧菌Vibrio mediterranei 117-T6（Vm 117-T6）对赤潮异湾藻、颗石藻、叉编金藻和东海原甲藻等微藻也具有较强的溶藻活性，可使其叶绿素含量显著降低，是一株广谱性溶藻细菌。Vm 117-T6能在40 min内使赤潮异弯藻的活动能力下降，光合作用受抑，感染120 min即可导致藻体裂解并产生白色絮状沉淀。Vm 117-T6能降解卡拉胶，不能降解果胶、纤维素和几丁质。菌体、胞内及胞外提取物均具有较强的溶藻活性，其胞外溶藻物质易溶于水、不溶于石油醚和乙酸乙酯，具较强极性；耐高温、不易被活性炭吸附、乙醇处理会降低溶藻活性。Vm 117-T6在常规与易感条件下的关键差异蛋白包括大量的ABC转运蛋白、外膜蛋白、鞭毛蛋白及少量Toxin。这些研究结果提示Vm 117-T6毒力效应物中含有极性非蛋白质类物质，且与毒力因子转运、分泌以及细胞黏附相关的蛋白可能在其毒力作用中发挥重要作用。上述研究结果表明：多种溶藻机制参与了Vm 117-T6的溶藻过程。     

茯苓多糖对中华鳖非特异性免疫功能的免疫调节作用

研究了口服茯苓多糖对中华鳖(Trionyxsinensis)非特异性免疫功能的影响。试验共分3个组,在基础饲料中添加20·0mg/kg茯苓多糖作为试验组,添加500·0mg/kg酵母细胞壁作为免疫刺激剂对照组,以投喂基础饲料作为空白对照组。连续投喂28d后,测定中华鳖血液中白细胞的吞噬活性、血清溶菌活力和补体活性。结果表明:在中华鳖饲料中添加茯苓多糖和酵母细胞壁能显著提高中华鳖白细胞的吞噬活性(t检验,P<0·05),而2个试验组之间中华鳖血液中白细胞的吞噬活性没有显著差异(t检验,P>0·05);2个试验组中华鳖血清中补体C3和C4的活性与对照组之间呈显著差异(t检验,P<0·05),而试验组之间无显著性差异(t检验,P>0·05)。本研究结果说明在中华鳖饵料中添加茯苓多糖和酵母细胞壁对中华鳖的非特异性免疫功能有增强作用。     

Evaluation of antioxidant properties and biofunctions of polar,nonpolar, and water-soluble fractions extracted from gonad and body wall of the sea urchin Tripneustes gratilla

This study was performed to compare the various bioactivities of water-soluble, polar, and nonpolar extracts from the edible part (gonad) and waste (body wall) of the sea urchin Tripneustes gratilla. During antioxidant activity screening, these three extracts from both gonad and body wall showed various antioxidant activities in different tests. During human skin fibroblast (CCD966SK) viability and collagen screening, only water-soluble and polar extracts from gonad and body wall were observed to promote cell viability, while the water-soluble gonad extract promoted the collagen-generating activity of CCD966SK cells. The water-soluble and polar extracts from gonad and body wall showed proliferative activity towards different cell types. In contrast, the nonpolar extracts of gonad and body wall exerted antiproliferative effects on most tumor cells. These results indicate that the bioactivities of sea urchin extracts depend on the part of the urchin that the extract was taken from as well as the extraction method used.     

Proteomic analysis of the sarcosine-insoluble outer membrane fraction of Flavobacterium columnare

Outer membrane proteins (OMPs) of bacteria are key molecules interacting with the host environment. Flavobacterium columnare, a pathogen-causing columnaris disease of fish worldwide, was studied in order to understand the composition of its OMPs. The sarcosine-insoluble membrane fraction of the OMPs was analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with reverse-phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC MS/MS). Thirty-six proteins were identified, including proteins involved in cell wall/membrane biogenesis, specific transport of various nutrients and in essential metabolism. The present study is the first report on the OMPs of F. columnare, and may serve as the basis for understanding the pathogenesis of the bacterium.     

Effects of protamine hydrochloride from chum salmon milt on lipid metabolism in rats

Protamine is a basic protein whose sulfate is used in medical applications as a carrier for injectable insulin and as a heparin antagonist, while its hydrochloride is used as an antibacterial ingredient for some food products. We previously showed that protamine hydrochloride derived from chum salmon milt inhibited pancreatic lipase and reduced plasma triacylglycerol levels in an oral fat tolerance test in humans. The present study evaluated the effect of chum protamine hydrochloride on lipid metabolism in rats. The single oral administration of protamine hydrochloride (500 mg/kg) reduced plasma triacylglycerol levels in a corn oil tolerance test, and repeated oral administration at a dose of 0, 150, 500, or 1,500 mg/kg with a high fat diet for 7 weeks significantly reduced body weight gain, liver weight, and epididymal adipose tissue, whereas total calorie intakes were not significantly different. These findings could be caused by the suppression of lipid digestibility leading to a significant increase in fecal lipid excretion due to the fact that protamine hydrochloride could bind with bile acids to make a complex that is difficult to digest. Protamine hydrochloride, with a considerable history as food, can be considered to be promising as an anti-obesity functional food material.     

拟诺卡氏菌骨架对凡纳滨对虾生长及免疫功能的影响

将研究室分离纯化的卢森坦拟诺卡氏菌扩大培养,发酵液经过超声波破碎、离心、胰蛋白酶酶解、脱脂、冷冻干燥,得到白色细胞壁骨架。在基础饲料中添加0.1%和0.2%的拟诺卡氏菌细胞壁骨架,饲喂凡纳滨对虾幼虾[体质量(4.21±1.28) g],养殖周期为4周,测定各处理组对虾体质量,统计各组存活率,使用南京建成试剂盒测定对虾血淋巴和肝胰腺组织中的一氧化氮合酶、超氧化物歧化酶、酚氧化酶、溶菌酶、谷胱甘肽过氧化物酶活性及丙二醛的含量,佛波豆蔻酸乙酯法检测对虾血淋巴中呼吸爆发活性,采用副溶血弧菌对剩余对虾进行攻毒,统计对虾死亡率。试验结果显示,拟诺卡氏菌细胞壁骨架能够明显促进凡纳滨对虾的生长,显著提高对虾血淋巴和肝胰腺中的超氧化物歧化酶、一氧化氮合酶、酚氧化酶、溶菌酶、谷胱甘肽过氧化物酶的活性( P <0.05, P <0.01),显著降低丙二醛浓度( P < 0.01 ),显著增强对虾血淋巴的呼吸爆发活性( P <0.05, P <0.01),可提高对虾的免疫保护率,从而提高凡纳滨对虾的存活率。研究结果表明,拟诺卡氏菌细胞壁骨架的适宜添加量为0.1%。     

Influence of several non‐nutrient additives on nonspecific immunity and growth of juvenile turbot,Scophthalmus maximus L.

The effects of three non‐nutrient additives on nonspecific immunity and growth of juvenile turbot (Scophthalmus maximus L.) were studied in this feeding experiment. The five treatments are basal diet alone, basal diets containing three different additives [0.4 g kg^?1 of xylo‐oligosaccharides (XOS), 1.3 g kg ^?1 of yeast cell wall and 0.8 g kg ^?1 of bile acids] individually or in combination. Two hundred and twenty‐five turbots (average initial weight 151.3 ± 11.3 g) were randomly allotted in five treatments with three replicates within each treatment in a 72‐day period. Comparing with basal diet group, activities of C3, C4, phagocyte, lysozyme, specific growth rate and feed conversion rate in yeast cell wall, XOS and the combined groups was enhanced significantly (P < 0.05); however, these parameters in bile acid groups were increased slightly (P > 0.05) except for phagocyte (P < 0.05); superoxide dismutase activity in additive groups was not significantly increased (P > 0.05) except for the combined group (P < 0.05). In conclusion, supplementation of yeast cell wall and XOS enhanced the nonspecific immunity of juvenile turbot. Synergistic or additive effect of the three additives was not observed.     

Antimicrobial activity of the biopolymer chitosan against Streptococcus iniae

The antimicrobial activity and mode of action of chitosan were evaluated against Streptococcus iniae, a pathogenic Gram‐positive bacterium of fish worldwide. Cell proliferation kinetics were examined following exposure to varying concentrations of chitosan. The action of chitosan on S. iniae was also investigated by measuring agglutination activity, conductivity, and extracellular and intracellular bacterial adenosine triphosphate (ATP) levels. Chitosan exhibited antibacterial activity against S. iniae at concentrations of 0.1% and above and was lethal at a concentration of 0.4% and higher. The mechanism of antibacterial activity of chitosan at the inhibitory level of bacterial growth appears to hinge upon the interaction between chitosan and the oppositely charged bacterial surface. This interplay causes agglutination, which was readily observed grossly and microscopically. After interacting with the cell surface via adsorption, an efflux of intracellular ATP was documented, which suggests that chitosan disrupts the bacterial cell causing leakage of cytosolic contents and ultimately cell death. Results suggest chitosan may be worth evaluating as a natural alternative to antibiotic against S. iniae infection of fish.