中文摘要：

采用快速膜乳化法制备粒径均一的聚（N-异丙基丙烯酰胺-co-丙烯酸）温敏微球,以其为载体构建新型固定化胰蛋白酶（Trypsin）;以牛血清白蛋白（BSA）为模型蛋白,分析了固定化Trypsin的酶解效果.结果表明,采用固定化Trypsin酶解30 min即可鉴定12条肽段,BSA氨基酸覆盖率为18%,优于自由酶酶解12 h的结果;固定化Trypsin在不同温度下的酶解效果差异显著,40～45℃的酶解效率较高,重复使用3次后,覆盖率为16%～18%,基本保持不变.

英文摘要：

A new type of immobilized trypsin was synthesized using uniform poly（N-isopropylacrylamide-co-acrylic acid）（Pc A） microspheres prepared by rapid membrane emulsification technique as carrier. Taking bovine serum albumin（BSA） as a model protein, the digestion performances of immobilized trypsin were investigated. Using a digestion time of only 30 min in immobilized trypsin, the protein was recognized with 12 peptides and the sequence coverage rate was 18%, it was better than that obtained by in-solution digestion. The digestion efficiency of immobilized trypsin was higher in temperature range of 40～45 ℃. In addition, immobilized trypsin also showed good durability and kept high activity after repeated use 3 times.