【摘要】 目的：以珍珠蚌肉为原料制备对二肽基肽酶（dipeptidyl peptidase,DPP）-IV具有抑制作用的生物活性肽。方法：利用两步酶解法并优化珍珠蚌肉酶解条件，筛选体外DPP-IV抑制活性较高组分进行超滤膜浓缩，SuperdexTM peptide 10/300 GL凝胶柱层析与反相高效液相色谱（reverse phase-high performance liquid chromatography,RP-HPLC）技术分离纯化出酶解液中的DPP-IV抑制肽，最终通过液相色谱-串联质谱鉴定其氨基酸序列。结果：中性蛋白酶与碱性蛋白酶对珍珠蚌肉均有良好降解效果，经3 kDa超滤膜浓缩后，酶解产物的DPP-IV抑制率可达67.4%。通过凝胶过滤柱和RP-HPLC进一步分离纯化出6个活性较高的肽段，序列分别为FNAPAM、FIPNY、IYNPPTPF、LAMPYP、FFVVMP和LAGMP。进一步，以分子对接的方式分析6个肽段与DPP-IV的相互作用关系。结论：本研究利用珍珠蚌肉制备DPP-IV抑制肽，可为以水产加工副产物为原料制备功能性食品提供理论参考。更多还原

【Abstract】 Objective: Bioactive peptides with an inhibitory effect on dipeptidyl peptidase-IV(DPP-IV) were prepared from pearl mussel muscle. Methods: Two-step enzymatic hydrolysis was adopted and enzymatic hydrolysis conditions were optimized. Fractions with in vitro DPP-IV inhibitory activity were concentrated by ultrafiltration, and purified by SuperdexTM peptide 10/300 GL gel filtration column chromatography and reverse phase-high performance liquid chromatography(RP-HPLC). DPP-IV inhibitory peptides were purified from the enzymatic hydrolysate and their amino acid sequences were identified by liquid chromatography-tandem mass spectrometry. Results: Both neutral protease and alkaline protease could effectively hydrolyze pearl mussel muscle. After concentration using a 3 kDa cut-off ultrafiltration membrane, the DPP-IV inhibition rate of the enzymatic hydrolysate reached 67.4%. After separation by gel filtration column and purification by RP-HPLC, 6 active peptides were obtained, whose sequences were FNAPAM, FIPNY, IYNPPTPF, LAMPYP, FFVVMP and LAGMP, respectively. Furthermore, the interactions between these peptides and DPP-IV were analyzed by molecular docking.Conclusion: Our present study provides a theoretical reference for the effective utilization of fish processing byproducts as raw materials for functional food production.更多还原