黄鱼鱼鳞胶原蛋白制备和纯化方法的研究

Methods of preparation and purification of collagen from Pseudosciaena polyactis scales

目的采用缓冲液-醋酸法，模拟中试条件，研究黄鱼鱼鳞胶原蛋白的分离和纯化方法。方法胶原蛋白的分离主要包括低温清洗、EDTA缓冲液浸泡、酸性缓冲液提取和透析纯化等4个工艺过程，用SDS-PAGE分析胶原蛋白的纯度，用差示扫描量热仪分析黄鱼胶原蛋白的热力学特性。结果采用该丁艺能获得纯净的黄鱼胶原蛋白。最佳工艺参数：浸提温度10℃以下、EDTA缓冲液（pH值7．5）浓度0．5mol／L、醋酸缓冲液浓度0．5mol／L、半透膜截流相对分子质量100000。纯化的黄鱼胶原蛋白在SDS-PAGE上呈现114000、124000、2000003条谱带，热变性温度为37．7℃。结论采用该法获得的黄鱼鱼鳞胶原蛋白可用于纯度要求较高的领域使用。

Objective To investigate the method of isolation and purification of collagen from Pseudosciaena polyactis scales with acetic buffer. Methods The preparation of collagen from Pseudosciaena polyactis scales mainly contained the following steps： washing scales with cool water to delete impurity; precipitating Ca^2＋ soaked by the EDTA buffer; extracting the collagen with acetic buffer; and dialyzing for purification. The purity of collagen was determined by SDS-PAGE and differential scanning cycler （DSC） was used to analyze the thermodynamic character. Results The optimum conditions in this technology were that extracted temperature should be controlled below 10℃ , concentration of the EDTA and acetic buffer were both 0.5 mol/L, and molecular weight over 100 000 were prevented by the dialysis-membrane. The prepared collagen showed three clear bands by SDS-PAGE, the molecular weights were 114 000, 124 000 and 200 000, respectively, the thermal denatured temperature was 37.7 ℃. Conclusions The isolated collagen from Pseudosciaena polyactis scales could be applied in the field demanding for higher quality.