摘要
为探索卵黄高磷蛋白肽(phosvitin phosphopeptide,PPP)对α-淀粉酶的抑制作用,使其能够调控血糖水平从而缓解Ⅱ型糖尿病,本研究以α-淀粉酶抑制率为指标酶解卵黄高磷蛋白制备PPP,利用酶抑制动力学实验分析PPP对α-淀粉酶的抑制类型,采用液质联用鉴定后用分子对接筛选出高活性的α-淀粉酶抑制肽,最后验证其活性。结果表明,最优酶解条件为先经胰蛋白酶(加酶量7000 U/g)再经胃蛋白酶(加酶量60000 U/g)分别酶解6 h,制得的PPP在浓度7.81×10-3 mg/mL时具有最高α-淀粉酶抑制率70.69%±1.71%。PPP对α-淀粉酶的抑制类型为混合型抑制。鉴定筛选出了两种新型的高活性α-淀粉酶抑制肽FGTEPDAK和IWGR,经验证其IC50值分别为(0.80±0.14)×10-3 mg/mL和(1.80±0.31)×10-3 mg/mL,极显著低于阳性对照阿卡波糖(3.17±0.47)×10-3 mg/mL(P<0.01)。本研究揭示PPP可作为新型降糖物质,应用于缓解Ⅱ型糖尿病的功能性食品开发中。
To explore the inhibiting effect of phosvitin phosphopeptide(PPP)onα-amylase,enabling it to regulate blood glucose levels and alleviate typeⅡdiabetes mellitus.This study employed enzymatic hydrolysis of phosvitin with the inhibition rate ofα-amylase as an index,followed by enzyme inhibition kinetics experiments to analyze the inhibitory type of PPP onα-amylase.LC-MS identification was conducted,and molecular docking was performed to screen for highly activeα-amylase inhibitory peptides,which were subsequently validated.The results showed that optimal enzymatic hydrolysis conditions involved initial hydrolysis with trypsin(7000 U/g),followed by pepsin(60000 U/g)for 6 h each.The prepared PPP exhibited the highestα-amylase inhibition rate 70.69%±1.71%at a concentration of 7.81×10-3 mg/mL.PPP acted as a mixed-type inhibitor.Two novel highly activeα-amylase inhibitory peptides FGTEPDAK and IWGR were identified and screened,exhibiting IC50 values of(0.80±0.14)×10-3 mg/mL and(1.80±0.31)×10-3 mg/mL,respectively.Both extremely significantly lower than the positive control acarbose's IC50 value(3.17±0.47)×10-3 mg/mL(P<0.01).This study highlights the potential use of PPP as a new hypoglycemic substance in developing functional foods for alleviating typeⅡdiabetes mellitus.
作者
薛天睿
吕彬斐
张铭冉
李灿
张晓维
XUE Tianrui;LÜBinfei;ZHANG Mingran;LI Can;ZHANG Xiaowei(College of Food Science and Engineering,Tianjin University of Science&Technology,Tianjin 300457,China)
出处
《食品工业科技》
CAS
北大核心
2025年第1期152-162,共11页
Science and Technology of Food Industry
基金
天津市自然科学基金(22JCYBJC01090)。
关键词
卵黄高磷蛋白肽
复合酶解
Α-淀粉酶抑制剂
分子对接
酶抑制动力学
phosvitin phosphopeptide
complex enzymatic hydrolysis
α-amylase inhibitors
molecular docking
enzyme inhibition kinetics
