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中国科学院大学学报 ›› 2004, Vol. 21 ›› Issue (3): 345-351.DOI: 10.7523/j.issn.2095-6134.2004.3.010

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Apocynaceae系细胞尿苷二磷酸葡萄糖焦磷酸化酶的纯化和表征

祁超   

  1. 中国科学技术大学生命科学学院结构生物学重点实验室, 合肥 230027
  • 收稿日期:2003-07-10 修回日期:2003-09-09 发布日期:2004-05-10
  • 基金资助:

    国家自然科学基金 (29701005);中国科学院重大项目 (KJ951A150402)资助

Purification and Characterization of UGPase

QI Chao   

  1. Key Laboratory of Structural Biology, School of Life Science, University of Science and Technology of China, Hefei 230027, China
  • Received:2003-07-10 Revised:2003-09-09 Published:2004-05-10

摘要:

经4步分离纯化的Apocynaceae植物培养细胞尿苷二磷酸葡萄糖焦磷酸化酶(UGPase)的比活为2049Umg,与细胞粗提液相比,活力提高97倍,活力回收率为21%,可直接用于尿苷二磷酸葡萄糖(UDPG)的合成,并且储存稳定性很好。UGPase的最适pH值为72,UGPase的最适温度为37℃左右。Mg2+是UGPase发挥活力所必需的。高浓度UTP和Glc6P均能抑制UGPase的活力。

关键词: 尿苷二磷酸葡萄糖焦磷酸化酶, 纯化, 表征, 比活

Abstract:

After isolation and purification of four steps, specific activity of UGPase from Apocynaceae cell is 204 9 U/mg. Compared with crude cell culture, specific activity is 97 times, and recovery yield is 21%. UGPase can be used in the synthesis of UDPG, and it has higher stability. The best available pH of UGPase is about 7 2, the best available temperature about 37℃. Mg2+ is necessary for UGPase to use. The high concentration of UTP and Glc 6 P can inhibit the activity of UGPase.

Key words: UGPase, purification, characterization, specific activity

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