环核苷酸磷酸二酯酶PDE9A的体外表达、纯化与酶活特性分析
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(1.北京工商大学 北京食品营养与人类健康高精尖创新中心/轻工科学技术学院, 北京 100048;2.北京科学仪器装备协作服务中心, 北京 100048;3.山东凯普菲特生物科技有限公司, 山东 日照 276800;4.日照华伟大健康产业研究院, 山东 日照 276800)

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国家自然科学基金面上项目(31972127);北京市教委科技发展重点项目(KZ201910011013)。


Expression, Purification and Enzyme Activity Analysis of Cyclic Nucleotide Phosphodiesterase PDE9A
Author:
Affiliation:

(1.Beijing Advanced Innovation Center for Food Nutrition and Human Health/ School of Light Industry, Beijing Technology & Business University, Beijing 100048, China;2.Beijing Scientific Instruments and Equipment Cooperation Service Center, Beijing 100048, China;3.Shandong KEEPFIT Biotech Co, Rizhao 276800, China;4.Rizhao HUAWEI Institute of Comprehensive Health Industries, Rizhao 276800, China)

Fund Project:

General Program of National Natural Science Foundation of China (31972127);Key Project of Science and Technology Development of Beijing Municipal Education Commission (KZ201910011013).

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    摘要:

    环核苷酸磷酸二酯酶PDE9A作为研究具有调节血糖等功效的天然活性物质的新型靶点,日益受到关注。通过异丙基硫代半乳糖苷诱导大肠杆菌BL21感受态细胞表达获得PDE9A,并通过Ni-NAT琼脂糖树脂亲和柱、Q-Sepharose离子交换纯化系统和Sephacryl S300分子筛纯化系统进行逐级纯化,最终获得高纯度的PDE9A蛋白,并应用高效液相色谱进行酶活特性检测,证明制备得到的蛋白质具有较高的水解cGMP能力,能够为新型降血糖功能性食品的开发提供依据。

    Abstract:

    As a novel target of blood glucose regulation, natural active substance cyclic nucleotide phosphodiesterase PDE9A was gradually attracted the attention. The target protein was obtained from E. coli BL21 cells via IPTG-induced expression. Then the protein was gradually purified by nickel column affinity purification system, Q-Sepharose ion exchange purification system and Sephacryl S300 molecular sieve purification system. Finally, the acquired target PDE9A protein with high purity was proved to have high affinity for substrate cGMP digestion, which provided a preliminary basis for the development of novel functional food for reducing blood glucose in the future.

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引用本文

王郅媛,李赤霞,张萌,王友升.环核苷酸磷酸二酯酶PDE9A的体外表达、纯化与酶活特性分析[J].食品科学技术学报,2020,38(5):85-90.

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  • 收稿日期:2019-10-14
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  • 在线发布日期: 2020-10-13
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