奥尼罗非鱼胶原蛋白及其酸解液美拉德反应产物的研究

以奥尼罗非鱼鱼皮中胶原蛋白为研究对象,采用氨基酸自动分析仪和GC-MS分析了胶原蛋白的游离氨基酸组成、主要风味成分及其酸解液美拉德反应产物的风味成分.结果表明：鱼皮中胶原蛋白的提取率为6.61%,酸解后的游离氨基酸组成与一般胶原蛋白的氨基酸组成一致,其中风味氨基酸占52.57%;美拉德反应产物中含3,3-二甲基正辛烷、2,6,10-三甲基十二烷等20种风味成分.     

鱼皮胶原蛋白肽复方制品对小鼠免疫功能的影响

本试验旨在研究鱼皮胶原蛋白肽与壳寡糖、人参提取物、黄芪提取物、当归提取物的药物组合物(鱼皮胶原蛋白肽复方制品)对免疫功能的影响。将336只SPF级健康ICR雌性小鼠随机分为四批(N=84),分别进行细胞免疫功能、脏器/体重比值及体液免疫功能、单核-巨噬细胞功能及NK细胞活性检测。每批随机分为7组(n=12),阴性对照组、高(3.0 g/kg)、中(1.0 g/kg)、低(0.5 g/kg)剂量复方制品组和鱼皮胶原蛋白肽组,连续灌胃30天后,测定各项功能指标。结果发现,与阴性对照组相比,各剂量组小鼠的迟发变态反应、脏器/体重比值、抗体生成细胞数无显著差异(P0.05);复方制品各剂量组小鼠的淋巴细胞增殖能力、血清溶血素水平以及单核-巨噬细胞碳廓清吞噬指数明显增强,且效果优于鱼皮胶原蛋白肽组。由此得出,鱼皮胶原蛋白肽复方制品具有较好的增强小鼠免疫力的作用。     

奥利亚罗非鱼胶原蛋白酸解液美拉德反应产物风味成分分析

以奥利亚罗非鱼鱼皮中胶原蛋白酸解液和葡萄糖为主要基质进行美拉德反应,采用氨基酸自动分析仪和GC-MS分析了游离氨基酸组成和主要风味成分。结果表明,酸解后游离氨基酸完全符合胶原蛋白特有的氨基酸组成,且风味氨基酸占59.77%。此外鉴定出19种风味成分,其中包括乙酸乙酯(30.34%)、2,3-二羟基丙醛、柠檬烯、1,1-乙二醇二乙酸酯、己二酸二丁酯、二乙二醇丁醚醋酸酯、苯甲酸乙酯、2-乙酰基吡咯等。     

基于串联质谱的鱼皮明胶鉴别研究

在胶原蛋白序列比对基础上,以虹鳟鱼明胶、猪明胶和牛明胶为模型,利用高效液相色谱－串联质谱技术（HPLC-MS/MS）研究了3种明胶降解多肽组成的差异。使用胰蛋白酶将鱼皮明胶进行了酶解处理,使用HPLC-MS/MS对酶解产物中的多肽组成进行了分析,并与猪和牛明胶酶解产物中的多肽进行了比较。结果表明鱼明胶酶解产物中存在特征多肽,通过特征多肽的种类可区别鱼明胶与猪和牛明胶,研究了明胶多肽中脯氨酸羟基化修饰、明胶分子量范围和脱酰胺化对特征多肽识别的影响。研究表明利用HPLC-MS/MS技术通过识别明胶酶解产物中的特征多肽进行鱼皮明胶鉴别具有可行性。     

产胶原蛋白酶嗜虫沙雷氏菌DL-12的紫外诱变育种

对产胶原蛋白酶的一株沙雷氏菌DL-12进行紫外诱变处理,以明胶为底物,对诱变后的菌种进行初步筛选,通过鱼皮液化实验和发酵上清液中的胶原蛋白酶活力的比较,选出紫外诱变不同剂量下产生的产酶活力有明显提高的菌株DL-12(50s)-3,DL-12(50s)-4和DL-12(55s)-5,对这3种菌的产酶稳定性进行测定和比较,其中诱变菌DL-12(50s)-4发酵上清液中酶活力平均达98.37U/m L,比出发菌提高了43.54%,且具有稳定产酶特性。由于经紫外诱变获得的诱变菌DL-12(50s)-4高产胶原蛋白酶且产酶稳定,是具有应用前景的资源菌种。     

大菱鲆鱼皮的氨基酸分析及评价

用常规方法测定了大菱鲆鱼皮的基本营养成分,用氨基酸自动分析仪分析其氨基酸构成,并对大菱鲆鱼皮中的氨基酸组成进行了营养学评价。结果表明,大菱鲆鱼皮蛋白质含量为14.8%,脂肪含量为1.8%,氨基酸含量为27.24%,其中含量较高的是甘氨酸、丙氨酸、脯氨酸和精氨酸,占总氨基酸的59.73%;必需氨基酸、甜味氨基酸、苦味氨基酸、鲜味氨基酸和涩味氨基酸的含量分别占总氨基酸的20.81%、60.61%、30.07%、10.43%和0.01%。甜味氨基酸、苦味氨基酸、鲜味氨基酸是大菱鲆鱼皮氨基酸的主要部分,构成了大菱鲆鱼皮的主要味道,大菱鲆鱼皮是一种口感柔滑、味道鲜美的理想补强食品。     

远去的赫哲鱼皮衣

<正>在我国北部黑龙江、松花江、乌苏里江流域居住着一个具有悠久历史而又勤奋的民族——赫哲族。赫哲族人民在几千年的发展过程中,创造了光辉灿烂的渔猎文化。他们不仅"食鱼肉",而且"衣鱼皮",成为世界上唯一以鱼皮为衣的民族,史称"鱼皮部"。赫哲族的鱼皮文化艺术是在其独特的渔猎生活中产生的,它是几千年来口传身授的民间活文化,是十分珍贵的     

双斑东方鲀鱼皮胶原多肽的制备及其在化妆品中的功效与刺激性评价

河豚鱼皮中富含胶原蛋白,是一种优质的胶原多肽制备原料。鱼皮的精深加工不仅能减缓资源的浪费也能扩大我国河豚鱼的加工模式。本研究以双斑东方鲀鱼皮为实验对象,通过单因素实验和正交实验制备得到双斑东方鲀鱼皮多肽(Fugu bimaculatus collagen peptide,FBCP)并对其功效性及皮肤刺激性进行了研究。结果显示,碱性蛋白酶制备FBCP的最佳工艺条件为固液比1∶10、酶解温度50℃、加酶量8 000 U/g、酶解pH值9.0、酶解时间4 h。在此条件下,FBCP的肽得率为39.65%。利用超滤膜将胶原多肽提取液分离成不同分子量的四种组分,分别为FBCP1(Mw1 kDa)、FBCP2(1 kDaMw5 kDa)、FBCP3(5 kDaMw10 kDa)和FBCP4(Mw10 kDa)。吸湿保湿试验中,FBCP1组分显示出优于其它组分的吸湿和保湿性能。体外抗氧化实验结果表明,FBCP1自由基清除活性最好,其对超氧阴离子O■、羟基自由基·OH和DPPH的IC_(50)分别为1.659、5.582和1.801 mg/mL。多次皮肤刺激和急性眼刺激实验表明,涂抹高达50%(W/V)FBCP对受试兔皮肤及兔眼均无刺激性。综上所述,FBCP具有良好的抗氧化和保湿功效且对皮肤和眼睛无刺激性,可为其在化妆品中的应用提供基础参考。     

牛蛙皮胶原蛋白酸提取条件优化及其对细胞生长和粘附性的影响

应用羟脯氨酸法测定了牛蛙皮胶原蛋白含量,通过正交实验对牛蛙皮胶原蛋白酸法提取条件进行了优化,并结合MTT法测定了胶原蛋白对细胞生长和粘附性的影响。结果显示,牛蛙皮胶原蛋白的含量约为45．1％;温度条件对提取率影响最大,其次依次为酸种、时间和浓度,最优组合为乙酸、1．5mol／L、37℃、24h;在低浓度时,牛蛙皮胶原蛋白对正常人类肝细胞的生长和粘附性无显著影响,当浓度升高到一定值时,对细胞生长和粘附性有显著促进作用。     

斑点叉尾鮰鱼皮提取物对鸡蛋涂膜保鲜效果的研究

斑点叉尾鮰鱼皮是加工生产中的副产物,其胶原含量丰富,附加值较高。本课题利用酶解法和水提法制备两种鱼皮提取液,分别处理A、B两组鸡蛋,于30℃下贮存,经分析,A组酶解物的主要成分三螺旋结构完整,为胶原蛋白;B组水提物中的主要成分三螺旋结构已破坏,保留一些β-转角结构和β-折叠结构,为胶原水解产物。通过定期检测鸡蛋的感官品质、失重率、相对密度、蛋黄指数和哈夫单位等指标判断其保鲜效果,结果表明,A、B两组保鲜效果显著优于空白组,且分别延长鸡蛋保质期4周和6周。     

Scale and bone type I collagens of carp (Cyprinus carpio).

1. Soluble Type I collagens were isolated from the scale and bone (skull) of lathyritic carp. Each tissue collagen was assumed to consist of two different molecular forms, (alpha 1)2 alpha 2 as a main component and alpha 1 alpha 2 alpha 3 as a minor one. 2. The possible coexistence of these two forms in soluble Type I collagen of carp was previously observed for skin and muscle, but not for the swim bladder in which only the form of (alpha 1)2 alpha 2 was found. 3. These composite results suggest the wide distribution of alpha 1 alpha 2 alpha 3 heterotrimers in collagenous tissues of carp.     

Proline hydroxylation of collagens synthesized at different temperatures in vivo by two poikilothermic species

Extent of prolyl hydroxylation in newly synthesized viper collagen is decreased at 10 degrees C to approximately 23% of normal on skin and to approximately 57% of normal in bone collagen. At 20 degrees C, prolyl hydroxylation is approximately 50% of normal in skin and normal in bone. At 10 degrees C and 20 degrees C, prolyl hydroxylation is decreased approximately 32% in the skin collagen of carp. In contrast, prolyl hydroxylation is unchanged at 10 and 20 degrees C in bone, scale and lepidotrichia. Prolyl hydroxylation of cartilaginous endoskeleton showed an approximately 25% decrease at 20 degrees C.     

Collagen from diamondback squid (Thysanoteuthis rhombus) outer skin

Collagens (acid-solubilized and pepsin-solubilized collagens) were prepared from diamondback squid outer skin and partially characterized. The yields of acid-solubilized and pepsin-solubilized collagens were about 1.3 and 35.6%, respectively, on a dry weight basis. Pepsin-solubilized collagen was heterotrimer with a chain composition of ala2a3. The patterns of peptide fragments were different from that of porcine skin collagen. Denaturation temperature was 27.5 degrees C, about 10 degrees C lower than that of porcine collagen. The amino acid composition of pepsin-solubilized collagen from diamondback squid outer skin was similar to that from cuttlefish outer skin. This squid is big among squid species, and its skin is thick. It is clear that diamondback squid outer skin has a potential as an alternative source of collagen to bovine skin and bone. At present, collagen using aquatic materials such as skin (cod and a deep-sea fish) and scale (sea bream and anchovy) is the development stage in the related industries. Unless the problem of BSE infection in land animals is resolved aquatic materials as an alternative source of collagen will attract much attention in the cosmetic and medical fields.     

Effect of temperature on dietary vitamin C requirement and lipid in common carp

We investigated the effect of temperature on the requirement of vitamin C in common carp. Small carp (0.70 g) was separately cultured at 25 and 35°C, respectively, and fed diet with or without supplement of 2000 ppm vitamin C. The fish increased weight gain and had smaller feed conversion rate at 25°C than at 35°C, with vitamin C-supplement and subsequently, without vitamin C-supplement. The level of vitamin C in the hepatopancreas and muscle, the ratio of hydroxyproline/proline and the level of collagen in the bone and skin were higher both at 25°C than at 35°C, with vitamin C-supplement and then without vitamin C-supplement. Judging from these results, the carp required vitamin C-supplement when cultured at high temperature. The level of thiobarbituric acid reactive-substance (TBARS) in the hepatopancreas and muscle of fish was higher at 35°C than at 25°C, without vitamin C-supplement and then with vitamin C-supplement. The level of glutathione was contrary to that of TBARS. Triglycerides and the ratio of polyunsaturated fatty acid (PUFA) in hepatopancreas and muscle of fish were higher at 25°C than at 35°C, and without vitamin C-supplement than with vitamin C-supplement. It is concluded that temperature and vitamin C influence lipid composition and peroxidation in the hepatopancreas and muscle of common carp.     

Altered ratio of collagen chains in bone of a patient with non-lethal osteogenesis imperfecta.

Bone from a patient with osteogenesis imperfecta contained type III collagen which was absent in control bone. The ratio of alpha 1(I)/alpha 2(I) in type I collagen of patient's bone was increased (2.9 vs. 2.3 +/- 0.2 in controls) and the ratio of dimers beta 11/beta 12/beta 22 was altered due to the increased beta 22 content. No abnormality was observed in collagen from the patient's skin. The altered composition of collagen in bone, but the normal composition in skin suggests that the disease in the patient is due to impaired regulation of the synthesis of collagens in bone, rather than by a mutation in one of the two type I collagen genes. Unlike in skin, all the type III collagen in patient's bone was pepsin-soluble indicating an inability of the bone to incorporate type III collagen into mature highly cross-linked extracellular matrix.     

Isolation and chemical characterization of collagen in bovine pulmonary tissues.

1. The contents of the fibrous proteins collagen and elastin in the pleural and parenchymal regions of bovine lungs were determined. The collagen content was approx. 70% (g/100g of salt-extracted defatted powder) in each tissue, and the elastin content was 28% in pleura and 13.5% in parenchyma. 2. Purification of the insoluble collagen from the pleura and parenchyma of bovine lungs by various methods was attempted. The collagen fractions isolated after incubation of the pulmonary tissues with the proteolytic enzymes collagenase ("collagenase-soluble" fraction) or pancreatic elastase ("elastase-insoluble" fraction) each contained approx. 87% of the total collagen initially present. 3. Both collagen fractions were chemically analysed for their amino acid and carbohydrate contents and were found to be similar to those of the intact interstitial collagens isolated from skin, bone and tendon. 4. The contents of the two aldimine cross-linking compounds, dehydrohydroxylysinonorleucine and dehydrodihydroxylysinonorleucine, were determined in the bovine pulmonary collagen fractions, and were found to decrease with increasing age of the animals, and were similar to the values found in intact collagens from bone and tendon.     

Separation of β22 dimer from bovine bone collagen

The precise role of the α₂-chain in collagen type I is of considerable scientific interest. Our recent studies demonstrated that the most noticeable difference between type I collagens, which were obtained from bovine hard tissues (bone, dentine) and soft tissues (tendon, skin), was presented in the position of β chain dimers using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The additional band observed both in the bone and dentine collagen was putatively identified as β₂₂ dimer (made of by an intermolecular cross-linking between two α₂-chains). Further investigations carried out on bovine bone and skin collagen, corresponding to hard tissue and soft tissue collagen respectively, confirmed this hypothesis. Successful separation of individual β₂₂ dimer from bone collagen was achieved. The procedure involves molecular-sieve chromatography on a Sephacryl S-400 column followed by differential acetone precipitation. Identification was done by the widely used methods, such as SDS-PAGE and cyanogen bromide (CNBr)-cleaved peptide analysis. It was proposed that the dimer and consequently α₂-chains may play important roles in the morphological and biological differences between hard and soft tissues.     

Molecular Consequences of the SERPINH1/HSP47 Mutation in the Dachshund Natural Model of Osteogenesis Imperfecta

Osteogenesis imperfecta (OI) is a heritable connective tissue disease characterized by bone fragility and increased risk of fractures. Up to now, mutations in at least 18 genes have been associated with dominant and recessive forms of OI that affect the production or post-translational processing of procollagen or alter bone homeostasis. Among those, SERPINH1 encoding heat shock protein 47 (HSP47), a chaperone exclusive for collagen folding in the ER, was identified to cause a severe form of OI in dachshunds (L326P) as well as in humans (one single case with a L78P mutation). To elucidate the disease mechanism underlying OI in the dog model, we applied a range of biochemical assays to mutant and control skin fibroblasts as well as on bone samples. These experiments revealed that type I collagen synthesized by mutant cells had decreased electrophoretic mobility. Procollagen was retained intracellularly with concomitant dilation of ER cisternae and activation of the ER stress response markers GRP78 and phospho-eIF2α, thus suggesting a defect in procollagen processing. In line with the migration shift detected on SDS-PAGE of cell culture collagen, extracts of bone collagen from the OI dog showed a similar mobility shift, and on tandem mass spectrometry, the chains were post-translationally overmodified. The bone collagen had a higher content of pyridinoline than control dog bone. We conclude that the SERPINH1 mutation in this naturally occurring model of OI impairs how HSP47 acts as a chaperone in the ER. This results in abnormal post-translational modification and cross-linking of the bone collagen.     

Genome-Scale Association Study of Abnormal Scale Pattern in Yellow River Carp Identified Previously Known Causative Gene in European Mirror Carp

Common carp (Cyprinus carpio) is one of the most widely studied fish species due to its great economic value and strong environmental adaptability. Scattered scale, a typical phenotype of the mirror carp that is derived from Europe, has never been observed in the Yellow River carp previously. We recently identified approximately one fourth of the F1 progenies displaying scattered scale in a full-sib Yellow River carp family in our breeding program, despite both parents that showed wild type with normal scale patterns. This family provides us unique materials to investigate the genetic basis underlying the abnormal scale mutant in Yellow River carp population. Genome-wide association study (GWAS) and association mapping were performed based on genome-wide single nucleotide polymorphisms (SNP) genotyped with common carp 250 K SNP genotyping array in 82 samples of the Yellow River carp family. We identified a 1.4 Mb genome region that was significantly associated with abnormal scattered scale patterns. We further identified a deletion mutation in fibroblast growth factor receptor 1 a1 (fgfr1a1) gene within this genome region. Amplification and sequencing analysis of this gene revealed a 311-bp deletion in intron 10 and exon 11, which proved that fgfr1a1 could be the causal gene responsible for abnormal scattered scale in the Yellow River carp family. Since similar fragment mutation with 306-bp and 310-bp deletions had been previously reported as causal mutation of scattered scale patterns in the mirror carp, we speculate that either the deletion mutation was introduced from Europe-derived mirror carp or the deletion independently occurred in the mutation hotspot in fgfr1a1 gene. The results provided insights into the genetic basis of scale pattern mutant in Yellow River carp population, which would help us to eliminate the recessive allele of the abnormal scale patterns in Yellow River carp population by molecular marker-assisted breeding.     

USE OF ULTRASONIC SPECTROSCOPY AND VISCOSIMETRY FOR THE CHARACTERIZATION OF CHICKEN SKIN COLLAGEN IN COMPARISON WITH COLLAGENS FROM OTHER ANIMAL TISSUES

Use of traditional sources of collagen such as pork, bovine, and carp has some limitations. Chicken skin can be valuable alternative. In this work collagen was isolated from chicken skin using a modified procedure. Molecular properties of chicken collagen were analyzed and compared to collagen from other animal skins. Acid-soluble collagen type I was obtained with a yield of 25% and water content around 67%. Viscosimetry and ultrasonic spectroscopy were newly used for molecular characterization. By ultrasonic attenuation measurements, a pre-aggregation phase in the interval from 20°C to 27°C was observed, which is a proof of disaggregation and liquefaction. From 40°C upward, the liquefaction process finishes and aggregation continues. In a bovine sample this phenomenon starts at 40°C, in chicken at 50°C, and continues until 70°C. By viscosimetry, the denaturation temperature was confirmed as 40°C for bovine and 50°C for chicken collagen. Chicken collagen has a two times higher lysine level than bovine, which provides molecular stability side-chain interactions. With regard to higher thermal stability and favorable amino acid composition, waste chicken skin has the potential to be an excellent alternative source of raw collagen with applications in the food industry and biomedicine.