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

1,2 1,3 1 1,4,*
WANG Zhiyuan ,摇 LI Chixia ,摇 ZHANG Meng ,摇 WANG Yousheng
(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)

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鄄induced expression. Then the protein was gradually purified by nickel column
affinity purification system, Q鄄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.
Keywords: PDE9A; exogenous expression; separation and purification; HPLC; enzyme activity analysis

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