食品科学 ›› 2011, Vol. 32 ›› Issue (19): 157-160.doi: 10.7506/spkx1002-6630-201119034

• 生物工程 • 上一篇    下一篇

微量热法研究10-23脱氧核酶切割反应的热动力学

周国燕,李炫辰,王爱民,蓝 浩,邢 华   

  1. 上海理工大学低温生物与食品冷冻研究所
  • 发布日期:2011-10-12
  • 基金资助:
    国家自然科学基金青年基金项目(50206013);上海市教委科研创新项目(09YZ230); 上海市重点学科建设项目(S30503);上海理工大学医疗器械与食品学院微创励志创新基金项目

Thermodynamics of 10-23 DNAzyme Cleavage Reaction Evaluated by Microcalorimetric Method

ZHOU Guo-yan,LI Xuan-chen,WANG Ai-min,LAN Hao,XING Hua   

  1. (Institute of Cryo-medicine and Food Refrigeration, Shanghai University of Science and Technology, Shanghai 200093, China )
  • Published:2011-10-12

摘要: 选用乙型肝炎病毒(HBV)前区C/C区2031位点的序列作为底物RNA片段,并合成相应的10-23脱氧核酶,然后用微量热法研究10-23脱氧核酶的热动力学。在近生理条件下,通过微量热法测定10-23脱氧核酶切割反应的热动力学参数。根据热流曲线得出底物RNA不同浓度的反应初速率v0,再作出v-[S]曲线,通过Lineweaver-Burk作图得出10-23脱氧核酶切割反应的最大速率vmax为2.5×10-9mol/(L ·s)、米氏常数Km为0.045nmol/L、催化常数Kcat为0.107min-1。

关键词: 10-23脱氧核酶, 切割反应, 微量热法, 热动力学

Abstract: The sequence of 2031 site in anterior HBV C/C area was selected as the substrate RNA fragment to synthesize the corresponding 10-23 DNAzyme. Under mimic physiological conditions, the thermodynamics of 10-23 DNAzyme was evaluated by microcalorimic method. Heat-flow curves were plotted to obtain the initial reaction rate (v0) at various concentrations of the substrate RNA. The maximum rate (vmax), Michaelis-menten constant (Km) and catalytic constant (Kcat) of 10-23 DNAzyme were obtained from the established Lineweaver-Burk plot of 1/v0 against 1/[S] to be 2.5 × 10-9 mol/(L ·s), 0.045 nmol/L and 0.107 min-1, respectively.

Key words: 10-23 DNAzyme, cleavage reactio, microcalorimetric method, thermodynamics

中图分类号: