云南鲷鱼皮酸溶性胶原蛋白的组成及理化特性

采用低温酸法提取云南鲷鱼皮酸溶性胶原蛋白(ASC),对ASC的氨基酸组成、亚基组成、结构特征、热稳定性以及溶解性等理化性质进行了研究,为云南鲷鱼皮的综合利用提供了理论依据。研究表明,ASC的主要氨基酸为甘氨酸、脯氨酸和丙氨酸,酪氨酸、蛋氨酸和半胱氨酸含量较低,脯氨酸的羟化率为35.47%;SDS-PAGE电泳图谱表明ASC主要由α_1、α_2以及β链组成,符合I型胶原蛋白的特征;ASC的最大紫外吸收波长为230 nm;红外光谱和X-射线图谱表明ASC分子排列规则紧凑,具有完整的三螺旋结构;差示热量扫描显示ASC的热变性温度分别为82.1和222.3℃,表明ASC的热稳定性较好;扫描电镜显示ASC分子呈三维的多片状结构,分布均匀,适合用作食品、药品和化妆品的载体和生物医学的基料;在pH值小于4或NaCl浓度低于40 g/L时,ASC具有良好的溶解性。     

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。     

水杨酸-金属与皮胶原蛋白热稳定性

通过研究水杨酸与3种金属离子(Cr(Ⅲ)、Al(Ⅲ)和Cu(Ⅱ))形成的Tanning Matrices(配合物-鞣性模块)与皮胶原蛋白作用,采用DSC测得的作用后皮胶原蛋白热变性温度Td,考察Tanning Matrices与皮胶原蛋白的耐热稳定性关系.结果证明两种物质形成Tanning Matrices与胶原蛋白作用的Td高于两者分别作用,同时发现,构成Tanning Matrices的配合物稳定常数越大,Td越高,即皮胶原蛋白耐热稳定性越高.     

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。     

秘鲁鱿鱼皮酸溶性胶原蛋白的分离及特征分析

以秘鲁鱿鱼皮真皮为原料提取酸溶性胶原蛋白(ASC)并对其性质进行研究。结果表明,秘鲁鱿鱼皮真皮ASC的羟脯氨酸含量为8.03%;SDS-PAGE分析表明其为I型胶原蛋白,至少含有2类亚基(α1和α2);紫外(UV)扫描光谱最大吸收波长为228 nm;红外(FTIR)光谱分析表明分子中存在三螺旋结构;热变性温度为32.0℃,比其他水产鱼类皮肤ASC高;相对溶解度p H2.0时最大,p H6.0时最小,分别为100%和8.57%;盐浓度达到2.0%以上时,相对溶解度迅速下降至4.23%。秘鲁鱿鱼皮可以作为一种提取酸溶性胶原蛋白的原料。     

鸡关节软骨Ⅱ型胶原蛋白结构及性能表征

从鸡关节软骨中得到的高纯度Ⅱ型胶原蛋白进行结构和性能的表征。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果显示仅有1条α带和1条β带,无其他杂蛋白带和I型胶原的条带,表明得到的Ⅱ型胶原纯度较高。紫外、红外和圆二色谱结果表明,该Ⅱ型胶原蛋白保持了完整的二级结构,即三股螺旋结构。氨基酸测定结果表明,Ⅱ型胶原含有较多的亚氨基酸,采用差示扫描量热法测得其变性温度高达40.2℃;Zeta电位法测得Ⅱ型胶原等电点偏弱酸性为5.06。     

鲢鱼鱼皮和鱼骨胶原蛋白的提取及理化性质分析

以鲢鱼鱼皮和鱼骨为原料,分别采用酸法和酶法从中提取酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(PSC),探讨其在氨基酸组成、等电点、热变性温度及黏度等方面的差异。结果表明:4种不同类型的胶原蛋白的紫外光谱扫描结果较为类似,在230 nm波长处均出现最大吸收峰,在280 nm波长处吸收峰不明显,氨基酸组成和比例大体类似,均符合Ⅰ型胶原蛋白的特征。鱼皮ASC、鱼皮PSC、鱼骨ASC、鱼骨PSC的热变性温度分别为30.5,31.7,31.2,32.1℃,等电点均在6.7左右。胶原蛋白的黏度随蛋白质量浓度的增加而增加;随Na Cl含量的增加而降低;p H值对胶原蛋白的黏度影响较大,在等电点附近时黏度最低,p H 3时黏度最大。     

预处理对皮粉微观形貌和热稳定性的影响

对极性溶剂、非极性溶剂、不同质子化程度、破坏氢键度及去氨基等预处理后的白皮粉的理化性能进行了表征。结果表明：尿素浓度在1~4mol/L范围内,皮粉纤维由未经任何处理的束状到表面粘连聚集再到纤维束分散,皮胶原的热变性温度急剧下降,浓度超过4mol/L后,纤维束则开始溃散,热稳定性随尿素浓度的增加呈缓慢降低趋势;有机酸及硫酸对胶原纤维的酸解、溶胀影响比盐酸的明显,甲酸处理后皮胶原的热稳定性降低较小,而硫酸对其酸解程度明显,热稳定性下降明显;去氨基后皮粉的纤维束也有一定程度的聚集,且在纤维表面有晶体析出,热分解温度变化甚微,而热变性温度降低显著。     

马面鱼皮胶原蛋白的提取及其特性研究

目的:从马面鱼皮中提取酸溶性胶原蛋白,并对其理化性质进行研究,为马面鱼皮的开发利用提供一定理论依据。方法:通过氨基酸分析、UV、FTIR及SDS-PAGE凝胶电泳初步探讨了酸溶性胶原蛋白的结构特征;同时对酸溶性胶原蛋白的等电点、热变性温度、热收缩温度及溶解性进行了研究。结果:氨基酸分析发现,马面鱼皮酸溶性胶原蛋白中甘氨酸含量为24.12%,羟脯氨酸含量为9.31%;每1 000个总氨基酸残基中,甘氨酸为349个,羟脯氨酸70个,未检测到色氨酸、半胱氨酸。UV光谱显示酸溶性胶原蛋白在210 nm波长处有最大吸收。FTIR光谱表明酸溶性胶原蛋白有特殊的三螺旋结构。SDS-PAGE显示酸溶性胶原蛋白有2条α链,1条β链和1条γ链;酸溶性胶原蛋白的等电点为5.70,热变性温度和热收缩温度分别为34.99℃和84.59℃。溶解性结果表明酸溶性胶原蛋白易溶于0.5 mol/L乙酸溶液,Na Cl含量1%~3%时溶解性稳定,之后随Na Cl含量增加,溶解性迅速下降。结论:马面鱼皮酸溶性胶原蛋白具有典型的Ⅰ型胶原蛋白特征,热稳定性较好,不同介质和盐含量对酸溶性胶原蛋白溶解度有很大影响。     

兔皮胶原蛋白的微观结构及不同因素对其聚集特性的影响

以兔皮胶原蛋白为原料,通过紫外光谱(UV)、红外光谱(FTIR)和扫描电镜(SEM)对兔皮胶原蛋白的微观结构进行了初步研究,在该基础上进一步研究了温度、胶原蛋白浓度、pH值和离子强度对其聚集特性的影响。紫外光谱和红外光谱的峰型及对应波长均符合Ⅰ型胶原蛋白的特征,电镜扫描观察到兔皮胶原蛋白为不规则的致密片状膜,部分表面褶皱;聚集特性研究表明,胶原蛋白浓度与测定温度升高,胶原蛋白的聚集速度和聚集程度均增加;在酸性环境中,胶原蛋白聚集时间较长;pH由7增至8的过程中,胶原蛋白的聚集程度先增加后降低,在pH 7.2时,聚集程度和速度均最高;随着离子浓度的增加,聚集程度和聚集速度先增加后降低,在NaCl浓度为120mmol/L时聚集速度最快,且自组装程度最高。     

酸法和酶法提取鳄鱼皮胶原蛋白及性质研究

对以鳄鱼皮为原料得到的酸溶性胶原蛋白(ASC)和酶溶性胶原蛋白(psc)的性质进行比较分析.紫外扫描结果表明所提取出的胶原蛋白在232nm波长处有显著吸收峰;SDS-PAGE结果表明ASC和PSC的肽链组成具有很大的相似性,均含有两种a肽链及其交联链(β链及γ链);溶解性分析表明鳄鱼皮胶原蛋白的等电点在pH7左右;ASC和PSC的保水性经过6h (25℃)仍然高于85％; ASC的吸油性(24mL/g)和PSC的吸油性(41 mL/g)差异较大.根据上述测定结果可知,鳄鱼皮胶原蛋白ASC和PSC组成类似,符合Ⅰ型胶原蛋白的特征,但二者具体的功能性质略有差异.     

COLLAGEN TYPES IN MECHANICALLY DEBONED CHICKEN MEAT

Mechanically deboned chicken meat (MDCM) from neck parts was digested with pepsin for 24 h at 4C. The solubilized collagens were subjected to salt fractionation both at acidic and neutral pH. The precipitates obtained after centrifugation, dialysis and lyophilization were quantitatively evaluated and collagen types were identified. The precipitate formed at 0.7 M NaCl pH 2.5, was dissolved in 1.0 M NaCl, 0.05 M-Tris-HCl, pH 7.5 buffer and further fractionated by sequential salt precipitation at 1.8, 2.5 and 4.5 M. Collagen types I, II, III and V were detected by electrophoresis techniques. Type I collagen was the major component. The presence of type II collagen indicated MDCM contained cartilaginous tissues.     

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。     

酸法和酶法提取牦牛骨胶原蛋白的特性分析

以牦牛骨为原料,分别采用酸法和酶法提取胶原蛋白并进行理化性质分析。通过氨基酸组成、紫外光谱、傅里叶变换红外光谱、热收缩温度、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gelelectrophoresis,SDS-PAGE）和扫描电子显微镜图像分析对胶原蛋白进行比较。结果表明：2?种方法的提取物都为典型的胶原蛋白,在230?nm波长左右出现最大紫外吸收峰;酸溶胶原蛋白（acid-soluble collagen,ASC）和酶溶胶原蛋白（pepsin-soluble collagen,PSC）的热收缩温度分别为40.12?℃和40.94?℃,变性焓值分别为0.25?J/g和0.22?J/g;红外光谱及SDS-PAGE分析表明,ASC和PSC主要由α、β、γ三种亚基组分组成,属于I型胶原蛋白,且三股螺旋结构完整;扫描电子显微镜结果表明,2?种方法提取的胶原蛋白都保留了较为完整的纤维网状结构,但酸法提取的胶原蛋白结构分布相对均匀。综合来看,2?种方法所提胶原蛋白在理化特性上并无太大差别,但酶法提取的胶原蛋白得率较高,酸法提取成本较低。     

Biochemical Characterization of Collagen in Myocommata and Endomysium Fractions of Carp and Spotted Mackerel Muscle

Myocommata and endomysium fractions were prepared from the different parts of body muscle of carp and spotted mackerel. Both type I and V collagens were detected in the myocommata and endomysium fractions of both fish. The relative concentration of type V collagen to type I collagen was higher in the endomysium fraction than in the myocommata fraction. Both type I and V collagens were less soluble in the endomysium fraction than in the myocommata fraction. These results indicated that the biochemical property of the collagen in the pericellular connective tissue was different from that of the collagen in the interstitial connective tissue of fish.     

Identification and Characterization of Molecular Species of Collagen in Fish Skin

ABSTRACT: Pepsin-solubilized collagen (PSC) prepared from the skin of 3 fish species—common horse mackerel, yellow sea bream, and tiger puffer—were separated into 2 fractions, major and minor, by ammonium sulfate precipitation. These collagen fractions were further purified by phosphocellulose column chromatography. From the results of SDS-PAGE, peptide mapping, and amino-acid analysis, the purified major and minor collagens were identified to be type I and V collagens, respectively. These results suggest that type V collagen might be widely present in fish skin as a minor collagen.     

Isolation and characterization of collagen from the cartilages of brownbanded bamboo shark (Chiloscyllium punctatum) and blacktip shark (Carcharhinus limbatus)

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) from the cartilages of brownbanded bamboo shark (BBS; Chiloscyllium punctatum) and blacktip shark (BTS; Carcharhinus limbatus) were isolated and characterized. ASC and PSC extracted from BBS cartilage showed the yields of 1.27 and 9.59 g/100 g (Based on dry weight), respectively, while yields of ASC and PSC from BTS cartilage were 1.04 and 10.30 g/100 g (based on dry weight), respectively. All collagens had protein as a major constituent with the trace amount of ash and fat. They contained glycine as the major amino acid with high contents of alanine, proline and hydroxyproline. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoretic patterns and subunit compositions, all collagens more likely comprised 2 types of collagen, type I and II, and contained α- and β-chains as the major components. Peptide maps of those collagens from both species digested by V8-protease and lysyl endopeptidase were different and were completely different from those of type I collagen from calf skin. Thermal transition temperature of ASC from those collagens (36.28-36.73 °C) was slightly higher than their corresponding PSC (34.56-35.98 °C). From zeta potential analysis, isoelectric points (pI) of collagen from the cartilages of BBS and BTS were estimated to be from 6.53 to 7.03 and from 6.96 to 7.26, respectively. Fourier transform infrared (FTIR) spectra of both ASC and PSC were quite similar, suggesting that pepsin hydrolysis did not affect the secondary structure of collagen, especially triple-helical structure.     

皮胶原的结构、性质与提取方法

简要介绍了动物皮胶原的结构组成和物化性质,着重综述了最近几年国内外从猪皮、牛皮和鱼皮中提取胶原的工艺方法。     

Temperature effects on type I pepsin‐solubilised collagen extraction from silver‐line grunt skin and its in vitro fibril self‐assembly

BACKGROUND: Fish skin is a potential source of collagen. Increasing the extraction temperature increases the yield of collagen. However, it may also result in degradation of the peptide chains, thus damaging the 3D structure of collagen that is vital for its application as a biomaterial. This work investigated the effects of extraction temperature on the yield and characteristics, including fibril self‐assembly, of type I pepsin‐solubilised fish skin collagen. RESULTS: Pepsin‐solubilised collagens were extracted from fresh skin of silver‐line grunt at 4, 10, 20 and 28 °C for 6 h. Extraction at 10 °C gave the highest yield of collagens (439.32 ± 96.43 mg g^?1 fresh skin, dry basis), which were identified as type I and comprised β, α1 and α2 subunits. Extraction at higher temperatures (20 and 28 °C) resulted in the formation of low‐molecular‐weight peptide fragments, thus reducing the yield of the resultant type I collagen. The denaturation temperatures of collagens extracted at 4 and 10 °C, as determined by thermal analysis using differential scanning calorimetry, were 39.5 and 37.5 °C respectively. In vitro fibril self‐assembly of 1 mg mL^?1 collagen solution (pH 6) incubated at 25 °C was only observed with collagens extracted at 4 and 10 °C. The 10 °C collagen not only showed a higher rate of self‐assembly, but its matrix also had a larger fibril diameter of 0.50 ± 0.07 µm (compared with 0.41 ± 0.07 µm for the 4 °C collagen) after 4 h of incubation. CONCLUSION: The results indicated strong effects of extraction temperature on the yield and characteristics of the collagen obtained. Extraction of pepsin‐solubilised collagen from silver‐line grunt skin at 4–10 °C gave a high yield of type I collagen with molecular integrity suitable for tissue‐engineering applications. Copyright © 2010 Society of Chemical Industry     

Comparison of physical–chemical properties of type I collagen from different species

Type I collagen is an important biopolymer and has been widely used in biomaterials due to its excellent biocompatibility and biodegradable properties. However, only a few studies have been reported on its comparison in different species. The amino acid composition, SDS-PAGE, UV–Vis spectrum, thermal transition temperatures, extractable uronic acid/protein ratio and enzymatic sensitivity of type I collagen from bird feet (BF), bovine skin (BS), frog skin (FS), porcine skin (PS) and shark skin (SS) were evaluated. The amino acid composition of type I collagens were different from different species, BF collagen contained higher glutamic acid (Glu) and aspartic acid (Asp), SS collagen contained lower aspartic acid and hydroxyproline (Hyp). Similar SDS-PAGE profiles were found from different animal’s collagen, all samples were composed of two α1-chain and one α2-chain. All UV–Vis spectrums exhibited a typical absorption peak at 218 nm. The UV absorption spectrum of BF collagen ranged from 190 to 340 nm, FS collagen ranged from 190 to 270 nm; the other species collagen ranged from 190 to 240 nm. Thermal transition temperatures of type I collagen from different animals decreased in the order of BF > BS > PS > FS > SS. PS collagen had higher extractable uronic acid/protein ratio and the lowest enzymatic sensitivity. Summarizing these results, the BF collagen had higher hyproxyproline (Hyp) + proline (Pro) value and exhibited higher thermal stability; the PS collagen contained larger amount of glycosaminoglycan and resulted in a high enzymes resistance. However, the BF and PS collagen should be used as a suitable material in biomaterial utilitys because of its better biostability.