牛源二肽基肽酶-IV的高效制备及性质

doi:10.7506/spkx1002-6630-20210805-069

食品科学 ›› 2022, Vol. 43 ›› Issue (16): 194-200.doi: 10.7506/spkx1002-6630-20210805-069

牛源二肽基肽酶-IV的高效制备及性质

赵嘉妮，陈宏，陈寅格，翁凌，张凌晶，张志刚，林家仕，曹敏杰

（1. 集美大学体育学院，福建厦门 361021；2. 集美大学食品与生物工程学院，福建厦门 361021；3. 肉食品安全生产技术国家重点实验室，福建厦门 361100）

出版日期:2022-08-25 发布日期:2022-08-31
基金资助:
“十三五”国家重点研发计划重点专项（2018YFD0901004）

Purification and Characterization of Dipeptidyl Peptidase-IV from Bovine Kidney

ZHAO Jiani, CHEN Hong, CHEN Yinge, WENG Ling, ZHANG Lingjing, ZHANG Zhigang, LIN Jiashi, CAO Minjie

(1. College of Physical Education, Jimei University, Xiamen 361021, China; 2. College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; 3. State Key Laboratory of Meat Production Safety Technology, Xiamen 361100, China)

Online:2022-08-25 Published:2022-08-31

摘要/Abstract

摘要： 以高效制备二肽基肽酶 IV（dipeptidyl peptidase-IV，DPP-IV）为目的，以牛肾为原料，经酸沉淀、硫酸铵盐析和凝胶过滤层析获得高纯度DPP-IV。还原条件下，纯化蛋白的分子质量为106 kDa，纯化倍数为169.9，得率为9.5%。对该蛋白进行质谱鉴定得到12 个肽段，共517 个氨基酸残基，与牛DPP-IV氨基酸序列的匹配度为100%。对其性质研究发现，DPP-IV为糖蛋白且以二聚体形式（210 kDa）存在，等电点为5.8。DPP-IV的二级结构具有典型的β-折叠构象，热变性温度为（72.7±0.3）℃。金属离子Zn2＋、Fe2＋及Fe3＋对DPP-IV活力有明显的抑制作用，1 mmol/L的Zn2＋对DPP-IV力抑制率高达98%。

关键词: 2型糖尿病；二肽基肽酶；分离纯化；性质分析

Abstract: The purpose of this study is to efficiently prepare high-purity dipeptidyl peptidase-IV (DPP-IV) from beef kidney by sequential acid precipitation, ammonium sulfate fractionation and gel filtration chromatography. Under reducing conditions, the molecular mass of the purified protein was estimated to be 106 kDa with a purification fold of 169.9 and a yield of 9.5%. Twelve peptides with a total of 517 amino acid residues were identified from the DPP-IV by mass spectrometry. The matching degree of the 12 peptides with the amino acid sequence of bovine DPP-IV standard was 100%. The DPP-IV was found to be a dimeric glycoprotein (210 kDa), with an isoelectric point of 5.8. The secondary structure of the DPP-IV had a typical β-sheet conformation, and the thermal denaturation temperature was (72.7 ± 0.3) ℃. Metal ions such as Zn2+, Fe2+ and Fe3+ revealed a significant inhibitory effect on the activity of DPP-IV. The inhibition rate of 1 mmol/L Zn2+ on the enzyme’s activity was as high as 98%.

Key words: type 2 diabetes; dipeptidyl peptidase-IV; purification; characterization

中图分类号:

Q556＋.3

赵嘉妮，陈宏，陈寅格，翁凌，张凌晶，张志刚，林家仕，曹敏杰. 牛源二肽基肽酶-IV的高效制备及性质[J]. 食品科学, 2022, 43(16): 194-200.

ZHAO Jiani, CHEN Hong, CHEN Yinge, WENG Ling, ZHANG Lingjing, ZHANG Zhigang, LIN Jiashi, CAO Minjie. Purification and Characterization of Dipeptidyl Peptidase-IV from Bovine Kidney[J]. FOOD SCIENCE, 2022, 43(16): 194-200.

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牛源二肽基肽酶-IV的高效制备及性质

Purification and Characterization of Dipeptidyl Peptidase-IV from Bovine Kidney

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