发酵核桃粕中有助于维持血糖健康水平肽的分析及降血糖机制  

Analysis of Peptides in Fermented Walnut Meal Contributing to Maintain Healthy Blood Glucose Levels and Underlying Hypoglycemic Mechanism

作　　者：刘莹莹 庞晓宇 郭聪聪 檀建新[1] 田益玲[1] LIU Yingying;PANG Xiaoyu;GUO Congcong;TAN Jianxin;TIAN Yiling(College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,China)

机构地区：[1]河北农业大学食品科技学院,河北保定071000

出　　处：《食品科学》2025年第6期38-46,共9页Food Science

基　　金：“十四五”国家重点研发计划重点专项(2022YFD1600402);河北省特色油料农业产业技术体系配套农机与产品加工专项(HBCT2024050206);河北省湿地保护与绿色发展协同创新中心项目(2023hbxczx1-11)。

摘　　要：为研究传统真菌发酵核桃蛋白产物中具有维持血糖健康水平的肽段,在发酵产物动物喂养总体评价的基础上,采用Sephadex G-50凝胶色谱和高效液相色谱分级分离纯化,以α-淀粉酶、α-葡萄糖苷酶和二肽基态酶-IV活性为评价指标,采用液相色谱-串联质谱进行分析,借助生物信息学工具对得到的分离组分IV、V、VI组共有的和差异的肽段,及其来源的蛋白,获得主要活性肽段,利用液相色谱测定含量,通过检索UniProt在线平台获得相关肽的来源,并用分子对接工具分析活性肽与3种酶的相互作用。结果表明,通过2型糖尿病小鼠造模,可以得到粗肽可以有效控制患病小鼠的体质重、摄食量和血糖,缓解胰岛素抵抗和提高葡萄糖耐量。在Sephadex G-50凝胶分离纯化后得到的分子质量较小的组分IV、V、VI中,组分V对α-淀粉酶和α-葡萄糖苷酶抑制活性与组分IV和VI没有显著差异,但是对二肽基态酶-IV抑制活性显著其他两组。随后对组分IV、V、VI进行多肽分析比较,并且对其相同肽段进行含量测定,发现在第V中,与VI相同的肽段MMRPDEDEQEGQRQ和LPLLR占总肽含量的82.3%,对DPPIV抑制活性94.3%源于这两条肽的作用。因此,在众多相同肽段中这两条肽链极具血糖调节活性肽开发前景。This study was undertaken in order to select peptides with blood glucose regulatory activity from fermented walnut meal with fungi.Animal feeding trials were conducted for overall evaluation of the crude peptide extract,which was subsequently fractionated by Sephadex G-50 column chromatography and high performance liquid chromatography(HPLC).The resulting fractions were tested for inhibitory activity againstα-amylase,α-glucosidase and dipeptidase-IV.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)and bioinformatics were combined to determine shared and differential peptides among fractions IV,V and VI and obtain the major active peptides,which were then quantified by LC.The proteins from which the active peptides were derived were identified by searching the UniProt online platform,and their interactions with the above three enzymes were analyzed by molecular docking.The results showed that the crude peptides effectively controlled body mass,food intake and blood glucose,mitigated insulin resistance and improved glucose tolerance in type II diabetic mice.Fractions IV,V and VI had low molecular mass,which were separated by Sephadex G-50 column chromatography.Fraction V showed no significant difference in inhibitory activity againstα-amylase andα-glucosidase but exhibited a significant difference in dipeptidase-IV inhibitory activity compared with fractions IV and VI.Comparative analysis showed that peptides MMRPDEDEQEGQRQ and LPLLR,which were found both in both V and VI,accounted for 82.3% of the total peptide content, and the peptides contributed to 94.3% of the inhibitory activity against DPP-IV. Therefore,these two peptides are very promising for glycaemic regulation.

关 键 词：核桃肽 降血糖 2型糖尿病小鼠 多肽序列 分子对接 

分 类 号：TS229[轻工技术与工程—粮食、油脂及植物蛋白工程]