酶膜耦合法制备均一分子量胶原蛋白多肽

Preparation of Collagen Polypeptide with Homogeneous Molecular Weight by Enzyme Membrane Coupled Method

为得到均一分子量胶原蛋白多肽,首先用固定化胰蛋白酶对胶原蛋白进行柱上循环酶切,经过3次柱上循环酶切后目标肽收率达到最高,含量较初始胶原蛋白提高70%以上;然后采用超滤法对酶解液进行分离,胶原蛋白多肽数均分子量达到15 600Da,具有良好的分子分布形态(M_(10)=8 300Da,M_(90)=21 000Da),目标肽总收率为23%,相对收率为174%。将制备的均一分子量胶原蛋白多肽用于交联型血浆代用品的制备,产品具有理想的体内应用理化指标。通过优化膜分离组件与酶解循环条件,可制备适用于不同原料来源和不同应用需求的均一分子量胶原蛋白多肽。

In order to obtain collagen polypeptide with homogeneous molecular weights,the collagen was cyclically digested with immobilized trypsin on a chromatography column.After three recycles of enzyme digestion,the yield of target peptide was the highest,and its content increased over 70% compared with the initial collagen.The enzymatic hydrolysate was further separated by an ultrafiltration method.The collagen polypeptide with Mn15 600 Da was prepared successfully,and it had good molecular weight distribution（M10=8 300 Da,M90=21 000Da）.The yield of target peptide was 23%,and relative yield was 174%.The prepared collagen polypeptide was used for the preparation of crosslinking plasma substitutes,and the products showed suitable physicochemical characteristics invivo.Through optimizing membrane separation module and enzymolysis cyclic condition,collagen polypeptide with homogeneous molecular weights applied to different material origins and utilizations could be prepared.