食品与发酵工业 >

2025 , Vol. 51 >Issue 22: 264 - 271

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042380

研究报告

火麻仁α-葡萄糖苷酶抑制肽的快速筛选、鉴定及抑制机理研究

  • 李焯颖 ,
  • 刘精杏 ,
  • 廖丹葵 ,
  • 李卫国 ,
  • 兰雄雕 ,
  • 刘彭如
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  • 1(广西民族大学 化学化工学院,广西多糖材料与改性重点实验室,化学与生物转化过程研究所,广西 南宁,530006)
    2(湖南省轻纺设计院有限公司,湖南 长沙,410015)
    3(广西大学 化学化工学院,广西石化资源加工及过程强化技术重点实验室,广西 南宁,530004)
    4(广西农业农村部农药研制与施用技术重点实验室,广西高工效农药及施用技术重点实验室,广西田园生化股份有限公司,广西 南宁,530007)
第一作者:硕士研究生(兰雄雕副教授和刘彭如讲师为共同通信作者,E-mail:lanxiongdiao@163.com;liu-pengru@gxmzu.edu.cn)

收稿日期: 2025-02-17

  修回日期: 2025-03-13

  网络出版日期: 2025-12-15

基金资助

广西高校中青年教师科研基础能力提升项目(2022KY0153);广西民族大学引进人才科研启动项目(2018KJQD06,2020KJQD12);广西自然科学基金面上项目(2021GXNSFAA075043);广西自然科学基金青年基金项目(2022GXNSFBA035499)

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Rapid screening, identification and inhibition mechanism of α-glucosidase inhibitory peptide from hemp seed

  • LI Zhuoying ,
  • LIU Jingxing ,
  • LIAO Dankui ,
  • LI Weiguo ,
  • LAN Xiongdiao ,
  • LIU Pengru
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  • 1(School of Chemistry and Chemical Engineering, Guangxi Minzu University, Guangxi Key Laboratory for Polysaccharide Materials and Modification, Institute of Chemical and Biological Transformation Processes, Nanning 530006, China)
    2(Hunan Light-Textile Industrial Design Institute CO.Ltd., Changsha 410015, China)
    3(School of Chemistry and Chemical Engineering, Guangxi University, Guangxi Key Laboratory of Petrochemical Resource Processing and Process Intensification Technology, Nanning 530004, China)
    4(Key Laboratory of Pesticide Development and Application Technology of Guangxi Ministry of Agriculture and Rural Affairs, Key Laboratory of High Efficiency Pesticide and Application Technology of Guangxi, Guangxi Tianyuan Biochemical Co.Ltd., Nanning 530007, China)

Received date: 2025-02-17

  Revised date: 2025-03-13

  Online published: 2025-12-15

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摘要

利用菠萝蛋白酶对火麻仁蛋白进行酶解,以α-葡萄糖苷酶抑制率为考察指标,通过单因素以及正交试验对酶解条件进行优化,得到最佳的酶解条件:酶解pH值为8.0,酶解温度为45 ℃,菠萝蛋白酶与蛋白质的比例为6 000 U/g,酶解时间为0.5 h。在该条件下,制备得到的酶解液对α-葡萄糖苷酶的半抑制浓度(half maximal inhibitory concentration,IC50)为(2.65±0.063) mg/mL。对酶解液进行超滤,收集<100 kDa的馏分,并使用生物复合材料3-MPBA/AG@aZIF-7/PDA对其进行亲和纯化,然后用LC-MC/MC质谱和计算机虚拟筛选,获得了5个具有潜在降血糖活性的新型多肽KFL、KFY、SLFK、SWG和YFK。体外实验和分子对接研究表明,KFY的活性最好[IC50为(3.10±0.22) mmol/L],5个多肽均能与α-葡萄糖苷酶上的氨基酸残基形成氢键和疏水作用,从而结合到α-葡萄糖苷酶上,形成稳定的复合物。该研究为火麻仁蛋白的高值化利用提供理论依据和数据支撑,同时也为天然降血糖药物的筛选提供参考。

关键词： 火麻仁; α-葡萄糖苷酶抑制肽; 计算机筛选; 分子对接

本文引用格式

李焯颖 , 刘精杏 , 廖丹葵 , 李卫国 , 兰雄雕 , 刘彭如 . 火麻仁α-葡萄糖苷酶抑制肽的快速筛选、鉴定及抑制机理研究[J]. 食品与发酵工业, 2025 , 51(22) : 264 -271 . DOI: 10.13995/j.cnki.11-1802/ts.042380

Abstract

Hemp seed protein was hydrolyzed by bromelain.Taking the α-glucosidase inhibitory rate as the evaluation metrics as the evaluation index, the enzymatic hydrolysis conditions were optimized through single-factor and orthogonal experiments, and the optimal enzymatic hydrolysis conditions were established as follows:the pH value for enzymatic hydrolysis was 8.0, the enzymolysis temperature was 45 ℃, the ratio of bromelain to protein was 6 000 U/g, and the enzymatic hydrolysis time was 0.5 h.Under these conditions, the semi-inhibitory concentration (IC50) of the enzymatic hydrolysate against α-glucosidase was (2.65±0.063) mg/mL.Ultrafiltration was carried out on the enzymatic hydrolysate to collect fractions less than 10 kDa.Then, the biological composite material 3-MPBA/AG@aZIF-7/PDA was used for affinity purification.Subsequently, through LC-MC/MC mass spectrometry and computer virtual screening, five novel polypeptides with potential hypoglycemic activity (KFL, KFY, SLFK, SWG, and YFK) were obtained.In vitro experiments and molecular docking studies showed that KFY had the best activity [(IC50 was (3.10±0.22) mmol/L], and all five peptides could form hydrogen bonds and hydrophobic interactions with the amino acid residues on α-glucosidase, thereby binding to α-glucosidase and forming a stable complex.This study provides theoretical basis and data support for the high-value utilization of hemp seed protein and also provides reference for the screening of natural hypoglycemic drugs.

Key words： hemp seed; α-glucosidase inhibitory peptide; computer screening; molecular docking

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