摘要
针对加工副产物鲍鱼外套膜利用率低的现象,对鲍鱼腹足和外套膜胶原蛋白相关性质进行比较研究,以期为鲍鱼的综合加工提供一定的理论依据。本研究以皱纹盘鲍为原料分别提取得到腹足酶促溶性胶原蛋白(pepsin-soluble collagen of abalone adductor,PSC1)和外套膜酶促溶性胶原蛋白(pepsin-soluble collagen of abalone mantle,PSC2),对PSC1和PSC2相关特性进行比较分析,并利用PSC1制备得到兔抗鲍鱼胶原蛋白多克隆抗体。SDS-PAGE显示,PSC1和PSC2分子组成均为(α1)3,且α1的分子量为140 ku,与水产无脊椎动物Ⅰ型胶原蛋白特征相似。对PSC1进行肽指纹图谱分析,获得6个肽段、含75个氨基酸残基,与盘鲍螺的胶原蛋白前肽α链和欧洲鲍螺的纤维状胶原一致性分别达100%和88%,证明纯化的PSC1为胶原蛋白。氨基酸组成分析表明,PSC1和PSC2的组成基本一致,但脯氨酸和羟脯氨酸含量均低于牛酸溶性胶原蛋白。圆二色谱分析结果显示,PSC1和PSC2溶液均在220和197 nm分别有一正、负峰,具有典型胶原蛋白三股螺旋结构特征。FTIR光谱分析结果提示PSC1和PSC2具有相似的三螺旋结构。利用兔抗皱纹盘鲍PSC1多克隆抗体对皱纹盘鲍、尼罗罗非鱼、鲤和仿刺参胶原蛋白进行免疫印迹分析发现,该抗体只与皱纹盘鲍PSC1和PSC2的α、β和γ链产生反应,表明该抗体具有良好的特异性。
Abalone mantle is the main by-product from abalone industry which is at a low utilization rate. The characterization of collagen from the abalone adductor and mantle was studied, for providing a theoretical foundation for processing of abalone. Purification and characterization of pepsin-soluble collagen from adductor（PSC1） and mantle（PSC2） of abalone（Haliotis discus hannai） were conducted. A polyclonal antibody was prepared against pepsin-soluble collagen from abalone adductor（PSC1）. The results of SDS-PAGE suggested that the structures of PSC1 and PSC2 were（α1）3 and the molecular weight of α1 chain was approximately 140 ku,which was the characteristic of typeⅠcollagen from aquatic invertebrates. Peptide mass fingerprinting（PMF） of PSC1 analysis obtained 6 peptide fragments including 75 amino acid residues which were identical with collagens from H. discus with 100% identity and H. tuberculata with 88% identity, suggesting PSC1 is collagen. Amino acid composition analysis showed that PSC1 and PSC2 had similar composition, while the amino acid content of them were lower than calf skin collagen by acid extraction. Circular dichroism（CD） spectrum analysis demonstrated a rotatory maximum at 220 nm and a negative peak at 197 nm of PSC1 and PSC2 solution, which were typical characteristics of the collagen triple helix structure. FTIR spectra further confirmed that both PSC1 and PSC2 have similar triple helical structure. According to Western blot analysis of collagens from 4 kinds of aquatic animals, the polyclonal antibody only positively reacted with α, β and γ chains of PSC1 and PSC2 from abalone, whereas no reactivity against collagens from Oreochromis niloticus, Cyprinus carpio and Stichopus japonicas was identified,indicating high specificity of the polyclonal antibody.
出处
《水产学报》
CAS
CSCD
北大核心
2016年第2期267-277,共11页
Journal of Fisheries of China
基金
国家自然科学基金(31471640)
国家科技支撑计划(2012BAD38B09)~~
关键词
皱纹盘鲍
酶促溶性胶原蛋白
圆二色谱
多克隆抗体
Haliotis discus hannai
pepsin-soluble collagen
circular dichroism
polyclonal antibody
