文献类型：学位论文

中文题名：军曹鱼鱼皮胶原蛋白的提取及其功能特性的研究

英文题名：Study on Isolation and Functional Properties of Collagens from the Skin of Cobia /(rachycentron Canadum/)

作者：杨树奇[1];

机构：[1]广东海洋大学;

导师：章超桦;广东海洋大学

授予学位：硕士

语种：中文

中文关键词：军曹鱼;酸溶胶原蛋白;酶促酸溶胶原蛋白;氨基酸组成;理化性质;功能特性

外文关键词：cobia; acid-solubilized collagen; pepsin-solubilized collagen; amino acid composition; physicochemical property; functional property

中文摘要：本研究以军曹鱼/(Rachycentron canadum/)皮为原料,分析测定其中的胶原蛋白含量,并在低温条件下利用酸溶解方法或酶促酸溶解方法提取酸溶性胶原蛋白/(ASC/)、酶促酸溶性胶原蛋白/(PSC/),并对其理化性质、亚基组成、功能特性进行研究,从而为军曹鱼皮的综合利用提供理论依据。研究结果表明: 军曹鱼鱼皮粗蛋白含量丰富,其含量为28.53/%/(湿基/),以干基计为81.58/%。军曹鱼鱼皮蛋白质组成中甘氨酸、丙氨酸、谷氨酸和脯氨酸含量最为丰富,含量高低依次为4.45、2.28、2.27、2.23/%,每1000个氨基酸残基中甘氨酸含量为328个,约占三分之一,符合胶原蛋白Gly-X-X的特点。胶原蛋白含量为20.09/%,胶原蛋白占其粗蛋白含量的72.4﹪。以干基计,军曹鱼鱼皮中ASC和PSC的提取得率分别35.5/%和12.2/%。 ASC、PSC氨基酸组成相似,甘氨酸含量极其丰富,分别占总氨基酸的32.9/%和34.2/%。ASC、PSC氨基酸组成中,每1000总氨基酸残基中亚氨基酸——脯氨酸和羟脯氨酸分别为213和197;ASC、PSC分别在219 nm、221 nm处有一强紫外吸收峰,符合Ⅰ型胶原蛋白光吸收特征;DSC曲线反映ASC、PSC的变性温度分别为38.17℃、36.03℃;ASC在pH 3时有最大溶解度,而PSC在pH 4溶解度最大;在pH 7时ASC、PSC溶解度最小,ASC在盐浓度大于2/%/(W//V/)时溶解度迅速下降,而PSC在盐浓度高于3/%/(W//V/)时溶解度迅速下降,当NaCl浓度大于2/%/(W//V/)时,PSC的溶解性优于ASC。这说明在酸性及低盐浓度的环境应用ASC、PSC更为方便。傅立叶红外转换扫描证明了实验制取的军曹鱼鱼皮胶原保存有比较完整的螺旋结构。 CM-52纤维素层析结果及SDS-PAGE电泳分析表明:ASC、PSC符合Ⅰ型胶原蛋白的结构特点,ASC、PSC均由3条α链组成,其组成形式为/(α1/)2α2、α1α2α3两种三聚体。SDS-PAGE电泳分析同时表明,α3链存在于军曹鱼皮胶原蛋白中。军曹鱼皮ASC和PSC的α1、α2的氨基酸分析结果表明,可以看出ASC和PSC的α1、α2链的氨基酸组成彼此非常相似但并不相同,这些数据表明ASC和PSC的α1、α2链均符合I型胶原的特征。 在吸湿性方面:在相对湿度/(RH/)43/%、81/%条件下, ASC和PSC的吸湿性要差于甘油,PSC要好于ASC。在体外保湿性方面:在RH=43/%、81/%下,0.2/% PSC、0.2/% ASC都能很好的降低失水率,说明具有良好的保湿性。在体表保湿性方面:所有的配方都较好的保湿效果,单纯PSC保湿性有好于丙二醇,单纯的ASC要比丙二醇的保湿性能差。而相对单纯的丙二醇而言,胶原蛋白和丙二醇的叠加效果更好。 胶原蛋白浓度达到0.25/%时,ASC与PSC的起泡性/(FA/)均达到最高。当pH为5时,ASC与PSC的FA均最高;不同盐浓度条件下,PSC比ASC的FA好;随着温度的变化,ASC和PSC的FA先提高然后就急剧下降,总体来说PSC要优于ASC。30℃时ASC和PSC的FA最好。不同pH条件下ASC与PSC的起泡稳定性/(FS/)相近,都具有较高的泡沫稳定性;随着NaCl浓度的增加,ASC、PSC的FS降低。30℃时ASC和PSC的FS最好。 ASC、PSC的乳化活性指数/(EAI随离子强度的增加而呈下降趋势,在0.25mol//L的氯化钠体系中,乳化性最好;ASC、PSC在酸、碱性溶液中具有较高的乳化能力,在等电点左右乳化能力达到低点;随着转速的增加、温度的提高、盐浓度的提高ASC、PSC乳化能力都有不同程度的提高。pH 4和pH 9时乳化稳定性最好;转速达到18000 r//min时,PSC乳化稳定性均达到最大;PSC在40℃时乳化稳定性最好,ASC的乳化稳定性对温度变化不明显。ASC和PSC具有较高的吸油性,ASC和PSC的吸油性分别为19.97 mL//g、21.65 mL//g。

外文摘要：In this study, collagens from cobia /(Rachycentron canadum/) skin were prepared, purified, and physicochemical characters, subunit compositions, functional characteristics were studied. Thereby, it provided theoretical basis for the comprehensive utilization of fish skin. The results were as follows: The skin of cobia was rich in the crude protein, which were up to 28.53/% on the basis of wet weight, however 81.59/% on the basis of dry weight. The glycin, alanine, glutamic acid and proline was most abundant in protein of cobia skin, contents were 4.45, 2.28, 2.27, 2.23/%, respectively. The amina acid composition was expressed as residues per 1000 total amino acid residues, the content of glycin was 32.8/%, approximately occurs uniformly, at every third residue throughout most of collagen molecules, which was consistent with the character of Gly-X-X. The content of collagen from the cobia skin was up to 20.09/% on the basis of wet weight. The content of collagen accounted about 72.4﹪of the crude protein. The yields of ASC and PSC were 35.5/% and 12.2/%, respectively, on the basis of dry weight. The amino acid composition of ASC and PSC were similar. The glycin was rich in ASC and PSC /(32.9/% and 34.2/%/), The amounts of imino acids, proline and hydroxyproline in ASC and PSC were 213, 197 residues per 1000 residues, respectively. A strong UV absorption peak were obtained at 219nm, 221nm,respectively. This result was in accordance with the characterization of the typeⅠ. The denaturation temperature of ASC and PSC were 38.17℃, 36.03℃from the curve of DSC, respectively, which was lower 5℃than that of calf skin collagen. Maximum solubility in 0.5M acetic acid was observed /(0.75 mg//mL and 0.78mg//mL/) at pH 3 and pH 4 for ASC and PSC, respectively. Solubility reached the minimum when the pH was 7. A sharp decrease in solubility was observed in the presence of NaCl, above 2/% and 3/% /(w//v/) for ASC and PSC, respectively. Reflectance Fourier transform infrared /(FT-IR/) microspectroscopy was applied to show relatively complete helix of collagens was preserved. Judging from chromatographic and electrophoretic patterns, ASC, PSC from cobia skin accorded with typeⅠcollagen, which were composed of threeαchains. Strongly suggesting the coexistence of /(α1/)2α2,α1α2α3 heterotrimers of type I collagen. An understanding of their functional difference must await further studies, because the isolation of theα1α2α3 heterotrimer had not been achieved so far. Tissue-specific existence ofα3 in cobia collagen was apparent. Judging from amino acid composition, the composition ofα1 andα2 of ASC and PSC was quite similar, but not identical. The moisture absorption of ASC and PSC was worse than glycerol, but PSC better than the ASC in temperature 25℃, under RH 43/%, 81/% conditions. In vitro, 0.2/% PSC, 0.2/% ASC could well reduce the water loss rate, indicating the good moisture. All the formulas were good in surface moisture retention, PSC was superior to propylene glycol, but more better than ASC. But to only propylene glycol, the moisturizing effect was much better by inputting ASC, PSC. When Collagen concentration was 0.25/%, the foamability of were the highest. The foamability was the highest in pH5. PSC was better than ASC in the foamability with indifferent salt conditions or temperature, in general. Both ASC and PSC arrived at highest at temperature 30℃. On different pH conditions, ASC and PSC were similar in having the higher foam stability. Along with the increase of NaCl, foam stability of ASC and PSC reduce. At temperature 30℃, it was the best. ASC and PSC's ESI decreased along with increasing the ionic strength, the best emulsifying capacity was found in 0.25mol//L of sodium chloride system. ASC and PSC’s emulsifying capacity was higher in acid or alkaline solution, but it was the lowest around the isoelectric point. Regardless of increasing rotational speed, enhancing temperature or raising the density of sodium chloride, the emulsifying capacity of ASC and PSC increased to varying degrees. At pH 4 and pH 9, emulsion stability were the best; ASC changed little in emulsion stability with speed increasing, when the speed was up to 18000 r//min, PSC reached the maximum of emulsion stability; the emulsion stability of PSC was the best at 40℃, but ASC did not change significantly. ASC and PSC had the higher oil absorption, high up to 19.97 mL//g, 21.65 mL //g, respectively.

年份：2010