辣根过氧化物酶的热稳定剂.PDF

生 物 工 程 学 报 Chin J Biotech 2009, March 25; 25(3): 388-391 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn © 2009 Institute of Microbiology, CAS CSM, All rights reserved 工业生物技术 1,2 2 2 2 1 毛新焕 , 李响 , 王姗姗 , 张文静 , 曾成鸣 1 , 710062 2 , 710062 : 保持酶的天然状态和高催化特性具有重要的意义。本研究筛选了辣根过氧化物酶(HRP)的稳定剂并研究了其作 用机制。结果发现硫酸镁和明胶能够显著提高HRP 的热稳定性, 并且两者具有协同作用。在硫酸镁和明胶组成的酶稳 定剂存在的条件下, HRP 在50oC 保温 80 h 后仍能保持 89%的活性, 常温下存放90 d 后可保持57%的活性, 而未加稳定 剂的对照样品中HRP 的残留活性分别为6%和小于 1%。通过对HRP 的Soret 带吸收光谱, 色氨酸内源荧光, ANS 荧光 进行分析, 揭示酶稳定剂可以明显降低在加热条件下HRP 的变性程度, 从而维持较为稳定的天然构象。 : 辣根过氧化物酶, 稳定剂, Soret 带吸收光谱, 内源荧光, ANS 荧光 Stabilizers of horseradish peroxidase 1,2 2 2 2 1 Xinhuan Mao , Xiang Li , Shanshan Wang , Wenjing Zhang , and Chengming Zeng 1 College of Chemistry and Material Sciences, Shaanxi Normal University, Xi ’an 710062, China 2 College of Life Sciences, Shaanxi Normal University, Xi ’an 710062, China Abstract: Keeping an enzyme in its native form with high catalytic activity is of great significance. In the present study, thermal stabilizers of horseradish peroxidase (HRP) were screened. The results indicated that thermal stability of HRP was enhanced by magnesium sulphate and gelatin. A synergic effect of magnesium sulphate and gelatin was observed. In the presence of the stabilizer, the enzymatic activity of HRP remained 89% after kept for 80 h at 50ºC and 57% for 90 days at room temperature. Thermal alterations of HRP structure in the absence an