不同鱼鳞胶原蛋白黏度特性研究

以四种鱼鳞为原料,用胃蛋白酶提取鱼鳞胶原蛋白,对胶原蛋白的分子量与不同条件影响下的黏度进行了研究。结果表明:同条件下制得的胶原蛋白分子量,鲤鱼主要分布约在94·0kDa以上,草鱼主要分布在约70·0kDa以上,鲫鱼主要分布约在65·0kDa以上,鲢鱼鱼鳞胶原蛋白分子量主要分布约在60·0kDa以上。鱼鳞胶原蛋白的黏度随着浓度的增加而增加;pH在3～4之间时,四种鱼鳞胶原蛋白黏度最大;NaCl与CaCl2对四种鱼鳞的胶原蛋白黏度的影响都呈现出相同的趋势,两者都使胶原蛋白黏度随其浓度增加而减小,但CaCl2对黏度的影响比NaCl的影响幅度变化大;四种鱼鳞胶原蛋白的黏度随丙三醇和乙醇的浓度增加而增加;鲢鱼胶原蛋白变性温度约为26℃,鲫鱼胶原蛋白变性温度约为23℃,草鱼胶原蛋白变性温度约为22℃,鲤鱼胶原蛋白变性温度约为24℃。     

鲢鱼鱼鳞胶原蛋白的流变特性

研究鲢鱼鱼鳞胶原蛋白的流变特性。采用MCR30高级流变仪和NDJ-8S数字式黏度计测定胶原蛋白溶液的流变特性值并绘制流变曲线。结果表明鲢鱼鱼鳞胶原蛋白溶液为典型假塑性流体;剪切速率、浓度和温度对胶原蛋白溶液的黏度影响较大;当振荡频率小于20 rad/s时,鲢鱼鱼鳞胶原蛋白溶液(1.0%)主要表现为黏性行为;当振荡频率大于20 rad/s时,鲢鱼鱼鳞胶原蛋白溶液(1.0%)主要表现为弹性行为;胶原蛋白发生热变性之后,其流体的弹性行为基本不变而黏性行为下降。     

一种以鱼鳞为原料的多产物联产工艺的建立与优化

鱼鳞中含有丰富的胶原和钙,可以作为生产胶原类产品和补钙产品的原料。为了提高鱼鳞的综合利用效果,解决鱼类加工过程中因副产物污染产生的环境问题,本研究以鱼鳞为原料,研究联合生产有机酸钙、胶原和胶原蛋白肽的工艺。具体地,鱼鳞先经过超声波辅助柠檬酸脱灰制备柠檬酸钙,再采用超声波辅助胃蛋白酶提取胶原,最后将提取胶原后的鱼鳞残渣用于制备胶原蛋白肽,并对所建立的联产工艺进行验证。结果表明,鱼鳞脱灰制备柠檬酸钙的条件为:超声波功率120 W,料液比1∶20 g/m L,柠檬酸浓度6%,处理时间180 min,草鱼鳞、鲢鱼鳞、沙丁鱼鳞的脱灰率分别为97.8%、97.6%、98.1%;胶原提取条件为:超声波功率180 W,料液比1∶20 g/m L,胃蛋白酶用量2000 U/g,处理时间8 h,草鱼鳞、鲢鱼鳞、沙丁鱼鳞胶原溶出率分别为40.1%、42.2%、56.2%;胶原制备条件为:底物浓度5%,胃蛋白酶用量280 U/g,初始p H2,处理温度65℃,草鱼鳞、鲢鱼鳞、沙丁鱼鳞胶原蛋白肽溶出率分别为91.7%、89.3%、88.9%。从鱼鳞中连续提取制备有机酸钙、胶原和胶原蛋白肽的工艺具有较好的可靠性和稳定性,有望在工业化生产中得到应用。     

鲢鱼鳞胶原蛋白的制备及性质研究

采用胃蛋白酶、木瓜蛋白酶和胰蛋白酶3种蛋白酶对鱼鳞中的胶原蛋白进行初步提取,选取效果最好的酶用正交实验优化胶原蛋白制备工艺。结果表明,用胃蛋白酶提取鱼鳞胶原蛋白效果最佳,最佳工艺条件为:40℃、48h、酶量4%、pH2,提取率为56.8%。经SDS-PAGE分析,所提取胶原蛋白分子量主要分布在30.0～94.0ku之间。氨基酸组成分析表明,鲢鱼鳞酶解蛋白液中甘氨酸(Gly)含量最高,脯氨酸(Pro)和谷氨酸(Glu)含量次之,而半胱氨酸(Cys)与酪氨酸(Tye)含量较低,符合I型胶原蛋白的组成特征。鱼鳞胶原蛋白随温度的升高黏度下降,乳化性次于对照的I型胶原蛋白。     

鲢鱼鱼皮中酸溶性胶原蛋白提取工艺研究

以鲢鱼鱼皮为原料,通过单因素试验和正交试验对鲢鱼鱼皮中酸溶性胶原蛋白的提取工艺参数进行分析,并用AR-500动态流变仪对鲢鱼鱼皮中酸溶性胶原蛋白的热变性温度进行分析。结果表明:鲢鱼鱼皮原料中酸溶性胶原蛋白提取的最佳工艺条件为:以乙酸作为提取剂,乙酸浓度0.3 mol/L,料液比1∶60(g/m L),提取时间96 h;此胶原蛋白的热变性起始温度为32.31℃,峰值温度为34.82℃。     

冬夏鲤鱼鱼皮和鱼骨胶原蛋白稳定性比较

从冬季(2008 年2 月)和夏季(2008 年7 月)的鲤鱼鱼皮和鱼骨中分别提取酸溶性胶原蛋白,在一定温度下加热,通过蛋白酶的酶解和圆二色谱(CD)法研究其热稳定性。结果显示：冬季鱼皮和鱼骨酸溶性胶原蛋白的变性温度分别为31.0℃和 32.8℃,比夏季鱼皮和鱼骨酸溶性胶原蛋白的变性温度分别低1.0℃和0.6℃。通过圆二色谱(CD)法得到了与此一致的结果。测定结果显示,来源于夏季鱼皮和鱼骨胶原蛋白的热稳定性要优于冬季鱼皮和鱼骨胶原蛋白。     

草鱼鳞和草鱼皮胶原蛋白性质及自聚集行为对比研究

以草鱼为原料,采取酸-酶结合法提取鱼鳞及鱼皮中胶原蛋白,经紫外光谱(ultraviolet spectrum,UV)、聚丙烯酰胺凝胶电泳(sodium dodecyl sulphate-polyacrylamide gel electrophoresis,SDS-PAGE)、傅里叶变换红外光谱(fourier transform infrared spectroscopy,FTIR)以及差示扫描量热法(differential scanning calorimetry,DSC)对比其结构性质,采用浊度实验、聚集程度实验和扫描电镜(scanning electronic microscopy,SEM)比较2种胶原蛋白的体外自聚集行为。结果表明,提取的2种胶原蛋白均具有2条α链,为典型I型胶原蛋白,并保持了良好的三螺旋结构,鱼鳞、鱼皮的变性温度分别为34.99℃和39.75℃。在30℃的中性盐溶液条件下,2种胶原蛋白均可产生自聚集行为且鱼皮、鱼鳞胶原蛋白的聚集程度分别为28%和27.33%。经SEM观察到2种来源胶原蛋白均能自聚集形成交织状纤维,其中鱼鳞胶原蛋白的网状纤维结构更加明显并具有胶原原纤维的周期性横纹D带,而鱼皮胶原蛋白自聚集的胶原纤维结构有一定坍塌且无D带。     

鲢鱼皮胶原蛋白的提取及性质分析

通过热水浸提,醋酸酸提及胃蛋白酶促提从鲢鱼皮中提取得到水溶性胶原蛋白(RSC),酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(PSC),并分析其部分性质。鲢鱼皮中RSC,ASC和PSC的得率以湿基计分别为13.2%,11.8%和13.6%;RSC,ASC和PSC的紫外光谱分析表明,其最大吸收峰都接近231 nm,但在280 nm处的吸收很小;傅立叶变换红外光谱结果表明鲢鱼皮胶原蛋白具有特殊的三螺旋结构;SDS-PAGE电泳图谱显示鲢鱼皮胶原蛋白有两条α链(α1和α2)和β链,表明该胶原为典型的I型胶原蛋白;鲢鱼皮RSC,ASC和PSC热变性温度分别为27.1℃,31.3℃和30.5℃。这些结果为进一步研究提供了理论依据。     

草鱼鱼鳞胶原蛋白的提取及其理化性质研究

以草鱼鱼鳞为原料,在低于蛋白变性温度的条件下提取酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(PSC),并对其理化性质进行系统测定。实验结果表明,ASC 和PSC 的热变性温度分别为30～35℃和28～35℃; PSC 在溶解性能、乳化性能和乳化稳定性方面优于ASC,而在溶液黏度、吸水性能、吸油性能、起泡性以及泡沫稳定性方面,ASC 明显好于PSC。     

尖吻鲈鱼鳞和鱼皮胶原蛋白的提取及其理化特性分析

以尖吻鲈鱼鳞和鱼皮为原料,提取并分离纯化酶溶性胶原蛋白,通过十二烷基硫酸钠-聚丙烯酰胺凝胶 电泳（sodium dodecyl sulfate-polyacrylamide gel electropheresis,SDS-PAGE）、氨基酸组成分析、差示扫描量热 （differential scanning calorimetry,DSC）、傅里叶变换红外光谱、X射线衍射和Zeta电位以及溶解度研究,分析 和比较了其主要理化性质。冷冻干燥后鱼鳞和鱼皮胶原蛋白得率（干质量）分别为2.3 g/100 g和47.3 g/100 g; SDS-PAGE结果显示两种胶原蛋白构型均为[α1(Ⅰ)]2α2(Ⅰ),初步判断属于Ⅰ型胶原蛋白;DSC结果显示鱼鳞和鱼 皮胶原蛋白热变性温度（Td）分别为37.54 ℃和36.74 ℃;傅里叶变换红外光谱和X射线衍射结果显示胶原蛋白经 胃蛋白酶处理后仍能保持其完整的三股螺旋结构;Zeta电位结果显示鱼鳞和鱼皮胶原蛋白等电点分别为pH 6.40和 pH 6.64;溶解度研究结果显示两种胶原蛋白在酸性条件和低NaCl质量浓度下均表现出良好的溶解性。     

鱼皮胶原蛋白性质的圆二色性和色谱法研究

从鲤鱼(Cyprinus carpio)和鳕鱼(Gadus morhua)鱼皮中提取酸溶性胶原蛋白(ASC),并对两种胶原蛋白的性质进行研究和比较。通过圆二色性(CD)分析鲤鱼鱼皮ASC的变性温度为36℃,鳕鱼鱼皮ASC的变性温度为19.0℃,与胶原蛋白亚氨基酸含量的分析结果一致。柱色谱层析结果显示,两种胶原蛋白都由3种不同的α 链组成(α1,α2,α3),3种单链形成的聚合体含有(α1)2α2 和 α1α2α3两种不同的形式。鲤鱼和鳕鱼鱼皮α1 链的胃蛋白酶酶解图谱显示两种胶原蛋白之间存在明显的差异。     

The effect of the subunit composition on the thermostability of collagens from the scales of freshwater fish

Collagens were isolated from the scales of carp and silver carp caught both in winter and summer. The denaturation temperatures measured by circular dichroism indicated that the thermostability of summer collagens was higher than that of the winter equivalents. The subunits of scale collagens were studied by column chromatography, which showed that the winter collagens had lower contents of α1 chain, but higher percentages of α2 and α3 chain, than the summer equivalents. It was suggested when season changes from summer to winter, the subunit compositions of scale collagens change accordingly; the content of α1 chain decreased, while contents of α2 and α3 chain increased. Therefore, the seasonal difference in the thermostability of scale collagens from the fish may be attributed to the subunit changes of collagens. This change may substantially affect tissue formation and construction, thus, have dramatic physiological consequences.     

Biochemical properties of bone and scale collagens isolated from the subtropical fish black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus)

Acid-soluble collagen (ASC) and pepsin-solubilized collagen (PSC) were isolated from the bones and scales of black drum (Pogonia cromis) and sheepshead seabream (Archosargus probatocephalus) caught in the Gulf of Mexico. ASC and PSC were analyzed for molecular weight by SDS–PAGE, amino acid composition, secondary structure, and denaturation temperature. The molecular masses of the collagen subunits were about 130 kDa for α₁ and 110 kDa for α₂, respectively. The amino acid composition of the PSCs was closer to that of calf skin ASC than to that of cod skin ASC. The melting temperatures of ASC and PSC were >34 °C. Intrinsic viscosity of the PSCs was similar to the intrinsic viscosity of collagen from fish species such as hake, cod, and catfish. Black drum and sheepshead bone and scale collagens were typical type-I collagens and may find applications in the functional food, cosmetic, biomedical, and pharmaceutical industries.     

草鱼鱼鳞、鱼皮和鱼骨酸溶性胶原蛋白特性对比研究

从草鱼鱼鳞、鱼皮和鱼骨中提取酸溶性胶原蛋白(ASC),对其特性进行对比分析。紫外光谱和电泳图谱结果显示,3种ASC紫外特征吸收峰出现位置与I型胶原蛋白紫外特征吸收峰出现的位置基本一致,分子结构都至少含有两条α链;傅里叶红外光谱显示,草鱼鱼鳞、鱼皮和鱼骨ASC在酰胺A带的N-H伸缩均以氢键形成缔合体,酰胺Ⅰ带和酰胺Ⅲ带的特征吸收峰表明鱼鳞和鱼骨ASC的α-螺旋结构保存较完整,鱼皮ASC的α-螺旋结构已经被破坏;3种ASC在热稳定性方面存在一定差异,热变性温度鱼骨ASC(34.5℃)鱼皮ASC(32℃)鱼鳞ASC(31℃),可能与胶原存在的组织和所处的环境有关。     

草鱼皮酸溶性和酶溶性胶原蛋白的提取及性质

本文研究了从草鱼皮中提取酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(PSC)及其部分性质。草鱼皮中ASC和PSC的提取得率以干基计分别为8.0%和18.6%;对草鱼皮ASC和PSC的紫外光谱分析,最大吸收峰都接近223nm;SDS-PAGE电泳图谱显示草鱼皮胶原蛋白是由两条不同的α链组成,分子量都在100kDa以上,与猪皮I型胶原蛋白相似;草鱼皮ASC和PSC热变性温度分别为33.8、34.5℃,只比猪皮的热变性温度(37℃)低3℃左右。结果表明草鱼皮胶原蛋白在功能食品、医药、化妆品、制药等方面有潜在的应用。     

鱼鳞胶原蛋白提取及抗氧化活性初探

比较鱼鳞胶原蛋白提取方法,观察其体内外抗氧化作用,为鱼鳞开发利用奠定基础。通过水提法、酸法及酸法与热水提取相结合的方法提取鱼鳞胶原蛋白,体外观察鱼鳞胶原蛋白对超氧阴离子(·O2-)、羟自由基(·OH)的清除作用。体内观察水提法胶原蛋白对小鼠血清、肝脏、大脑中SOD、CAT含量的影响。结果表明:水提法及酸法与热水相结合的方法得到的胶原蛋白提取率明显优于酸法的提取率,3种方法得到的胶原蛋白对·O2-、·OH均具有良好的清除作用,并呈一定的剂量依赖关系;水提法得到的胶原蛋白能明显提高小鼠血清、肝脏、大脑中的SOD、CAT含量。水提法可很好提取鱼鳞胶原蛋白,且得到的胶原蛋白具有显著的抗氧化生物活性。     

Biochemical properties of skin collagens isolated from black carp (Mylopharyngodon piceus)

Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were extracted with the yield of 15.5 and 26.5% on the basis of dry weight from the skin of black carp (Mylopharyngodon piceus). ASC and PSC were similar in amino compositions and subunit compositions, but they were slightly different in the secondary structure. Both of ASC and PSC, either rehydrated in distilled water or in acetic acid, had a lower denaturation temperature than pig skin collagen (PPSC), but their denaturation temperature were higher than that of collagens from other freshwater fish. Intrinsic viscosity of different collagens decreased in the order of PPSC>PSC>ASC. Both of ASC and PSC showed only partial degradation in collagenase solution, but they revealed more sensitive to collagenase compared with PPSC. Black carp skin collagens did not induce a significant cytotoxic effect according to the results of in vitro cytotoxicity test.     

Study on the properties of gelatins from skin of carp (Cyprinus carpio) caught in winter and summer season

The objective of this work was to compare the physiochemical (molecular weight distribution and amino acid composition) and rheological (viscosity property, gel strength and melting point) properties of gelatins from skins of carp caught in winter to those obtained for the summer equivalents. Gelatins from winter and summer fish skins were extracted at 60, 70 and 80 °C. SDS-PAGE patterns for gelatins extracted under the same conditions showed that the degradation of gelatins from winter fish skins were more severe than that of the summer ones. The imino acid contents of the winter and summer gelatins extracted at 60 °C were very similar, showing 190 and 188 residues/1000 residues, respectively. The gelatins from summer fish presented higher melting points and gel strengths, as well as better viscosity properties than the winter equivalents (P < 0.05). The differences in the rheological properties between winter and summer gelatins may be explained by different thermostability of interstitial collagen molecules (from which gelatins were derived) in the two seasons.