许氏平鲉鱼皮胶原蛋白与胶原肽的制备及特性研究

目的从许氏平鲉鱼皮中制备胶原蛋白及胶原肽,并对其特性进行初步探讨。方法通过氨基酸组成、傅里叶变换红外光谱(FTIR)、十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)及扫描电镜分析探讨鱼皮胶原蛋白的结构特征;制备不同分子量胶原肽组分SCH 1(Mw2000 U)与SCH 2(Mw2000 U),测定其起泡性、吸水性及对H2O2诱导L929细胞氧化损伤的保护作用。结果氨基酸分析表明鱼皮胶原蛋白亚氨酸含量为16.7%;FTIR表明样品基本保持了天然三螺旋结构;扫描电镜分析显示冷冻干燥胶原蛋白具有多孔状结构。大分子量SCH 1的起泡性较好,而低分子量SCH 2具有较好的吸湿性。SCH对H2O2引起的细胞损伤产生一定的保护作用,并呈现出剂量效应关系,结论许氏平鲉鱼皮胶原蛋白为I型胶原蛋白,其胶原肽具有良好的起泡性与吸湿性,可有效抵御H2O2引起的细胞氧化损伤。     

发酵法制备罗非鱼皮胶原蛋白肽的工艺的研究

目的:研究利用枯草芽孢杆菌进行发酵法生产罗非鱼皮胶原蛋白肽的制备工艺。方法:以罗非鱼皮和水为原料,接入枯草芽孢杆菌进行发酵,对罗非鱼鱼皮浓度、装液量、接种时间、接种量、发酵时间进行了考察,通过单因子实验和正交实验优化确定了最佳发酵工艺。结果:最佳发酵工艺为:罗非鱼鱼皮含量为3%,装液量为90mL/250mL,接种量为5%,接种时间为14h,发酵时间为52h。并且在此发酵工艺下,罗非鱼鱼皮蛋白的水解度高达36.88%。结论:采用枯草芽孢杆菌发酵法制备罗非鱼皮胶原蛋白肽,工艺简单、成本低、不污染环境、水解度高,是值得推广的制备胶原蛋白肽的方法。     

Angiotensin I-converting enzyme inhibitory activity of low-molecular-weight peptides from Atlantic salmon (Salmo salar L.) skin

Collagen extracted from Atlantic salmon (Salmo salar L.) skin (which is normally discarded in the process of manufacture) was hydrolyzed with Alcalase and papain, and treated by multistage separation. The salmon skin collagen peptides (SSCP) obtained had high protein content (91.20 ± 1.03%) and low molecular weights, 90.79% of which were less than 1000 Da. SSCP was then separated by reversed-phase high performance liquid chromatography. Eleven major fractions were collected and their angiotensin I-converting enzyme (ACE) inhibitory activity was assayed. Fractions 5 and 7 displaying higher ACE inhibitory activity were subjected to mass spectrometer to identify the ACE inhibitory peptides. A total of eleven peptide sequences were identified, and two dipeptides, Ala-Pro and Val-Arg, were selected for further ACE inhibitory activity analysis. The ACE inhibitory activities of Ala-Pro (IC₅₀ = 0.060 ± 0.001 mg/ml) and Val-Arg (IC₅₀ = 0.332 ± 0.005 mg/ml) were found to be approximately 20- and 4-fold higher than that of SSCP (1.165 ± 0.087 mg/ml), respectively.     

乳源蛋白中的活性肽及其生理作用

乳源蛋白了营养作用外，还具有生物活性作用，这是由于其中存在着生物活性肽的缘故。这些活性作用包括：阿片活性、等作用。叙述了乳源蛋白中的活性肽及其生理作用。     

具有自由基清除活性的鱿鱼皮胶原蛋白肽酶解工艺研究

以蛋白水解度和羟自由基清除率为指标,采用胰蛋白酶法提取秘鲁鱿鱼皮中的胶原蛋白肽,并通过单因素试验和正交试验优化了鱼皮胶原蛋白肽的制备工艺条件,同时测定了鱼皮的基本组成成分.结果表明,加酶量5％,酶解时间7h,酶解温度50℃,pH值8.0,酶解效果最好.此条件下,鱿鱼皮胶原蛋白肽的水解度为26.1％,羟自由基清除率为83.3％,在医用保健食品和抗衰老美容护肤产品上具有很好的实用价值.     

应用于食品工业的活性肽及其生产方法

本对应用于食品工业的活性肽的功能、应用及来源进行了简要介绍。对活性肽的几种生产方法即酶解法、化学合成法、酶合成法、DNA重组技术进行了综述，并对这几种生产方法的优缺点进行了评述。     

In vitro skin permeation and retention of parabens from cosmetic formulations

Parabens are antimicrobial agents widely used in foods, cosmetics and pharmaceutical products. Although non-mutagenic, non-teratogenic and non-carcinogenic, parabens can induce allergic contact dermatitis and posses estrogenic activity. The aim of this work was to assess the skin permeation and retention of methyl- (MP), ethyl- (EP) and propyl- (PP) paraben from three commercial cosmetic creams. The results obtained indicate that parabens are capable of permeating through and accumulating in the skin. The extent of penetration depends more on paraben characteristics (solubility, lipophilicity) than on the composition of the formulation. In particular, the percentage permeated across the skin was independent of the composition of the cream used and decreased in the order MP, EP and PP, in accordance with decreasing solubility. After 8 h of contact with the skin, 60% of MP, 40% of EP and 20% of PP were found across the skin. Concerning skin retention, the percentage remaining in the skin after 8 h depends on both paraben characteristics and on the composition of the formulation used. In conclusion, it appears that only the type of paraben, in particular its water solubility, affects skin penetration whereas the composition of the emulsion, which influences skin retention, plays a secondary role. Finally, excised rabbit ear skin can be considered as a good model for human skin for in vitro experiments.     

大鲵皮胶原蛋白肽的结构特性及其对乙醇诱导肝损伤小鼠的保护作用

研究了大鲵皮胶原蛋白肽结构特性及其对乙醇诱导肝损伤小鼠的保护作用。大鲵皮经胃蛋白酶水解制备胶原蛋白,经木瓜蛋白酶水解制备大鲵皮胶原蛋白肽。紫外光谱及红外光谱结果表明,大鲵皮胶原蛋白肽具有胶原蛋白的三股螺旋结构。SDS-PAGE结果表明大鲵皮胶原蛋白肽分子量低于55ku。大鲵皮胶原蛋白肽低（50mg/（kg·d））、高（150mg/（kg·d））剂量组可显著抑制小鼠乙醇诱导产生的血清中AST、ALT活性的升高,显著抑制肝组织MDA含量升高,并且抑制肝组织中SOD活性的下降。组织病理表明,乙醇诱导肝损伤模型组小鼠肝细胞明显肿胀变形,肝细胞索紊乱,伴有炎症细胞侵润,大鲵皮胶原蛋白肽各剂量组的肝细胞排列规则、肝细胞索清晰。结果表明,大鲵皮胶原蛋白肽具有保护乙醇诱导小鼠肝损伤的作用。      

鱼皮胶原蛋白及胶原活性肽的研究

近年来,鱼皮胶原蛋白及其活性肽的研究越来越多。不同提取法得到的胶原蛋白的功能性质具有较大差异,而胶原蛋白的功能性质与其结构紧密相关。本文比较了几种主要的胶原蛋白提取方法,探讨了胶原蛋白结构的研究进展,较详细的阐述了鱼皮胶原蛋白、胶原活性肽的应用及其研究前景。      

响应面法优化罗非鱼鱼皮胶原多肽螯合镁的工艺条件的研究

以罗非鱼鱼皮为原料,研究鱼皮胶原多肽和镁离子的螯合工艺条件。探讨了鱼皮胶原多肽与氯化镁的质量比、反应体系的pH、螯合温度、螯合时间等因素对鱼皮胶原蛋白螯合反应的影响;在单因素实验的基础上,通过响应面法对鱼皮胶原多肽与镁的螯合工艺条件进行优化,确定最佳的螯合工艺条件为:质量比1∶5、pH7.0、螯合温度70℃、螯合时间30min。在此条件下,螯合率达84.58%。通过对比螯合反应前后傅里叶变换红外光谱图的变化,证明了鱼皮胶原多肽与镁离子形成了新的螯合物。      

脱色工艺对鱼皮胶原肽品质的影响

利用复合酶通过酶解方法制备罗非鱼皮胶原肽,考察活性炭、大孔吸附树脂对胶原肽的脱色效果。以脱色率和肽损失率为指标,通过正交试验进行工艺优化,结果表明:在pH 4.5、活性炭添加量2%(m/V)、脱色时间60min、蛋白浓度50mg/mL的条件下,活性炭脱色效果最佳,脱色率可达92.66%,肽损失率为11.23%;而大孔吸附树脂在最佳脱色条件下,脱色率只有83.02%,肽损失率却高达30.77%,脱色效果明显不及活性炭。根据胶原肽分子量分布和氨基酸组成分析结果发现:经活性炭、大孔吸附树脂脱色后,鱼皮胶原肽中分子量500~1 000Da组分比例明显减少,His、Tyr和Phe等氨基酸含量显著下降。     

两种来源于牛皮胶原蛋白的新型ACE抑制肽

从牛皮胶原蛋白中鉴定分离出抑制血管紧张素Ⅰ转换酶（ACE）的多肽片段。首先优化牛皮胶原蛋白水解方法,应用胃蛋白酶和碱性蛋白酶获得活性最高的水解产物（抑制ACE的IC50为0.168 mg/m L）。经MW 5000超滤分离后,得到水解液中活性最高的部分（IC50为0.079 mg/m L）。对分离后低于MW 5000的部分经RP-HPLC进一步分离,发现含有大量ACE抑制肽,并应用RP-HPLC-MS/MS鉴定出两种新型ACE抑制肽,氨基酸序列分别为ISVPGPM和LGPVGNPGPA,IC50值分别为0.017 mg/m L（24.3μmol/L）和0.077 mg/m L（87.8μmol/L）。最后,本文模拟了两种抑制肽在体内生理环境下的消化,发现均可被部分降解,且消化产物具有与原始多肽相近的ACE抑制活性。      

Oral administration of marine collagen peptides from Chum Salmon skin enhances cutaneous wound healing and angiogenesis in rats

BACKGROUND: A wound is a clinical entity which often poses problems in clinical practice. The present study was aimed to investigate the wound healing potential of administering marine collagen peptides (MCP) from Chum Salmon skin by using two wound models (incision and excision) in rats. RESULTS: Ninety‐six animals were equally divided into the two wound models and then within each model animals were randomly divided into two groups: vehicle‐treated group and 2 g kg^?1 MCP‐treated group. Wound closure and tensile strength were calculated. Collagen deposition was assessed by Masson staining and hydroxyproline measurement. Angiogenesis was assessed by immunohistological methods. MCP‐treated rats showed faster wound closure and improved tissue regeneration at the wound site, which was supported by histopathological parameters pertaining to wound healing. MCP treatment improved angiogenesis and helped form thicker and better organised collagen fibre deposition compared to vehicle‐treated group. CONCLUSION: The results show the efficacy of oral MCP treatment on wound healing in animals. Copyright © 2011 Society of Chemical Industry     

大菱鲆鱼皮抗氧化胶原肽的制备及特性分析

以大菱鲆(Scophthalmus maximus)鱼皮为原料制备抗氧化胶原肽。进行了鱼皮酶解的单因素实验、酶解条件的响应面优化、G-15葡聚糖凝胶层析分离及体外抗氧化活性检测。结果表明：大菱鲆鱼皮酶解的最适用酶为中性蛋白酶,最佳工艺条件为：酶解温度52 ℃,酶解时间为338 min,加酶量为1142 U/g,反应体系pH值为6.7。按照此最佳条件制备得到鱼皮酶解液羟自由基清除率为34.44%,与理论值接近。大菱鲆鱼皮胶原肽经过层析分离,得到了3种不同分子质量的多肽组分TSP1、TSP2、TSP3,其中TSP3体外抗氧化活性最好,具有更强的羟自由基清除能力、亚铁离子螯合能力、DPPH自由基清除能力以及还原力。本研究为大菱鲆及其它鱼类多肽的开发研究以及鱼类副产物高值化利用提供了技术参考。     

鱼鳔胶原肽抗氧化稳定性研究

以DPPH自由基、OH自由基清除能力和还原力为评价指标,探究pH、温度、食品配料、金属离子、防腐剂以及体外模拟胃、肠道消化对鱼鳔胶原肽抗氧化稳定性的影响。结果表明:鱼鳔胶原肽在酸性环境中抗氧化活性保持得较好,在碱性条件下丧失较快;鱼鳔胶原肽具有较强的耐热性,高温不会降低其抗氧化活性;蔗糖、葡萄糖以及防腐剂(苯甲酸钠和山梨酸钾)对鱼鳔胶原肽的活性影响不明显,但添加高浓度的食盐会降低鱼鳔胶原肽的抗氧化活性;金属离子Zn~(2+)、Cu~(2+)对鱼鳔胶原肽抗氧化活性的影响较大,Ca~(2+)的影响较小,K~+、Mg~(2+)无显著影响;模拟胃肠道消化对鱼鳔胶原肽抗氧化活性无显著影响。     

Preparation,isolation and hypothermia protection activity of antifreeze peptides from shark skin collagen

Antifreeze proteins have characteristics of inhibiting the growth of crystals, decreasing the injury of cells and can retain the structure, texture and quality of productions. The purpose of this study is to obtain natural, safe, high activity antifreeze peptides using various separating means, and to detect their hypothermia protect activity on Lactobacillus bulgaricus. Collagen derived from shark skin was hydrolyzed with Acid Protease. Then the hydrolysate was subsequently added on to SP-Sephadex C-25 column, Sephadex G-50 gel filtration column and C18 Reversed-phase high performance liquid chromatography to acquire a pure component. A purified hydrophilic peptide, named SsC-AFP, with the characteristic of pI < 5.0 and molecular weight of 906 Da, was obtained. The amino acid sequence of SsC-AFP was GAIGPAGPLGP, which consisted of Gly–Pro–x tripeptide repeat sequence. When the concentration of SsC-AFP was 250 μg/mL, the survival rate of L. bulgaricus was up to 90.28%.