Study on Isolation, Identification and Activity of Novel α-Glucosidase Inhibitory Peptide Derived from Dahe Black Pig Ham(大河乌猪火腿中新型α-葡萄糖苷酶抑制肽的分离、鉴定及活性研究)
摘要
Protein degradation during fermentation and maturation of Dahe black pig ham results in abundant bioactive peptides. In order to investigate whether there were α-glucosidase inhibitory peptides in Dahe black pig ham and its activity, ham peptides of different molecular masses were prepared by ultrafiltration separation, and α-glucosidase inhibition rate was used as an index to identify, screen and study the activity of peptides in Dahe black pig ham by peptidomics combined with bioinformatics analysis. The results showed that the peptide of Dahe black pig ham with molecular weight less than 3kDa had good α-glucosidase inhibitory activity. A total of 143 peptides were identified from Dahe black pig ham, mainly derived from myosin, troponin and β-enolase. The further screened peptide IEEALGDK showed a good α-glucosidase inhibitory activity (<i>IC</i><sub>50</sub>=1.42mg/mL). The results of BIOPEP-UWM search revealed that the peptide IEEALGDK was a novel bioactive peptide. Peptide stability studies have shown that the IEEALGDK peptide has good thermal stability, acid and alkali resistance and gastrointestinal digestive stability. Molecular docking results showed that the peptide IEEALGDK mainly exerted its active effects by occupying the active residue sites of α-glucosidase, Arg594, Arg727, Arg799 and Arg467 through hydrogen bonding and hydrophobic interactions. The IEEALGDK peptide derived from Dahe black pig ham had good α-glucosidase inhibitory activity and the study aimed to provide a theoretical support for the further development and use of the Dahe black pig ham peptide.(大河乌猪火腿在发酵和后熟过程中蛋白质降解可产生丰富的生物活性肽。为探究大河乌猪火腿中是否存在α-葡萄糖苷酶抑制肽及其活性,以大河乌猪火腿为研究对象,通过超滤分离方法制备了不同分子质量的火腿肽；以α-葡萄糖苷酶抑制率为指标,采用肽组学结合生物信息学分析方法对火腿肽进行鉴定、筛选和活性研究。结果表明:分子质量小于3 kDa的大河乌猪火腿肽具有良好的α-葡萄糖苷酶抑制活性。从大河乌猪火腿中共鉴定出143条主要来源于肌球蛋白、肌钙蛋白和β-烯醇化酶的肽序列,进一步筛选出的肽段IEEALGDK具有良好的α-葡萄糖苷酶抑制活性(<i>IC</i><sub>50</sub>值为1.42mg/mL)。BIOPEP-UWM数据库检索结果显示,肽段IEEALGDK为新型的生物活性肽。肽的稳定性研究表明,肽段IEEALGDK具有较好的热稳定性、耐酸碱性和胃肠道消化稳定性。分子对接结果显示,肽段IEEALG