摘要
为从鲟鱼皮中分离提取出二肽基肽酶-IV(dipeptidyl peptidase IV,DPP-IV)抑制活性肽,以鲟鱼皮胶原蛋白为主要材料,利用蛋白酶进行酶解,以DPP-IV抑制率为主要考察指标,在单因素试验的基础上,进一步利用响应面法优化了鲟鱼鱼皮中DPP-IV抑制肽的制备工艺条件,确定最佳酶解条件为:酶解温度50℃、料液比1∶100(g/mL)、pH 6.12、加酶量10170.35 U/g、酶解时间12.12 h。在此基础上,通过超滤、凝胶层析及反相高效液相色谱的方法对酶解产物进行分离纯化;采用液相色谱-串联质谱法对多肽进行鉴定,结果显示纯化后具有DPP-IV抑制活性肽氨基酸组成为:甘氨酸-脯氨酸-丝氨酸-甘氨酸-亮氨酸-天冬氨酸-甘氨酸-丙氨酸-赖氨酸(GPSGLDGAK),IC_(50)值可达(61.27±1.16)μmol/L。本研究结果可为利用鲟鱼皮生产新型的生物活性成分提供参考。
In order to prepare dipeptidyl peptidase IV(DPP-IV)inhibitory peptide,sturgeon skin collagen was hydrolyzed by protease.A combination of one-factor-at-a-time method and response surface methodology was used optimize the preparation conditions based on percentage of DPP-IV inhibition.The optimal enzymatic hydrolysis conditions were determined as follows:temperature 50℃,solid-to-liquid ratio 1:100(g/mL),pH 6.12,enzyme dosage 10170.35 U/g,and enzymatic hydrolysis time 12.12 h.The hydrolysate prepared under the optimized conditions was separated and purified by sequential ultrafiltration,gel filtration chromatography and reverse-phase high performance liquid chromatography(RPHPLC).As identified by liquid chromatography-tandem mass spectrometry(LC-MS/MS),the amino acid composition of the purified peptide with DPP-IV inhibitory activity was GPSGLDGAK,and its half-maximum inhibitory concentration(IC_(50))value was(61.27±1.16)μmol/L.The results of this study can provide a reference for the production of new bioactive components from sturgeon skin.
作者
尹剑
武瑞赟
胡锦蓉
李平兰
YIN Jian;WU Ruiyun;HU Jinrong;LI Pinglan(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2022年第6期195-203,共9页
Food Science
基金
北京市鲟鱼鲑鳟鱼创新团队项目(BAIC08-2021)。
关键词
鲟鱼皮
二肽基肽酶-IV抑制活性肽
酶解法
sturgeon skin
dipeptidyl peptidase IV inhibitory peptide
enzymatic hydrolysis
