光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

引用本文: 王玮, 洪军, 黄琨, 肖保林, 杨卫云, 赵莹雪, 高云飞. 光谱和电化学方法研究自组装纳米簇模拟过氧化物酶[J]. 分析化学, 2012, 40(10): 1543-1548. doi: 10.3724/SP.J.1096.2012.20208 shu

Citation: WANG Wei, Hong Jun, HANG Kun, YANG Wei-Yun, XIAO Bao-Lin, ZHAO Ying-Xue, Gao Yun-Fei. Investigation of Self-assembled Nano-cluster Artificial Peroxidase by Voltammetry and Spectroscopy[J]. Chinese Journal of Analytical Chemistry, 2012, 40(10): 1543-1548. doi: 10.3724/SP.J.1096.2012.20208 shu

光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

通讯作者: 洪军

基金项目:
本文系河南大学特聘教授科研启动基金(No.5443D08010) (No.5443D08010)

伊朗国家科学基金资助

摘要: 利用细胞色素c与Nafion,通过自组装方式,构建了纳米簇模拟过氧化物酶;利用电子透射显微镜、圆二色谱仪、紫外可见光分光光度仪及电化学工作站等设备测量模拟酶结构变化、酶促动力学参数以及电化学特性;在50 mmol/L Na₂HPO₄-NaH₂PO₄缓冲溶液(pH 10.0)中,测得模拟酶米氏常数K_m、催化速率及催化效率分别为2.5 μmol/L、0.069 s^-1、0.028±0.005 (μmol/L)^-1s^-1,其催化效率为天然辣根过氧化物酶的39%±5%,该模拟酶具有很好的稳定性和活性,能够替代辣根过氧化物酶。该模拟酶可以固定在功能化多壁纳米管与纳米金复合材料修饰玻碳电极上,在50 mmol/L Na₂HPO₄-NaH₂PO₄缓冲溶液(pH 7.0)中测得模拟酶与电极之间电子迁移速度常数k_s为0.65 s^-1。利用循环伏安法测量模拟酶催化浓度为0.02-10 nmol/L的H₂O₂,其阴极峰电流随H₂O₂浓度增加而增大,并且得出模拟酶催化H₂O₂检出限为0.05 nmol/L,电化学表观米氏常数K_mapp为2.3×10^-10mol/L。

关键词:

自组装
/ 纳米簇
/ 模拟酶
/ 光谱学
/ 电化学

English

Investigation of Self-assembled Nano-cluster Artificial Peroxidase by Voltammetry and Spectroscopy

1.
College of Life Science, Henan University, Kaifeng 475000, China

Corresponding author: 洪军

Received Date: 31 March 2012
Fund Project:
本文系河南大学特聘教授科研启动基金(No.5443D08010)

伊朗国家科学基金资助

Abstract: A nano-cluster with highly efficient peroxide activity was constructed self-assembly based on nafion (NF) and cytochrome c (Cyt c), The UV-Vis spectrometry, Circular Dichroism (CD) and transmission electron microscopic (TEM) methods were utilized for additional characterization of the artificial peroxidase (AP). The nano-cluster was composed of chain-ball structure, with an average ball size of approximately 40 nm detected by TEM method. The Michaelis–Menten (Km) and catalytic rate (kcat) constants of the AP were determined to be (2.5±0.4) μmol/L and (0.069±0.001) s^-1, respectively, in 50 mmol/L PBS at pH 7.0. The catalytic efficiency of AP was evaluated to be (0.028±0.005) μmol/L^-1s^-1, which was 39%±5% as efficient as the native HRP. AP generated here can be used in place of HRP with high stability and activity. The AP was also immobilized on a functional MWNCTs-gold nano-complex modified glassy carbon electrode. The cyclic voltammetry of AP on the nano complex modified glassy carbon electrode showed a pair of well-defined quasi-reversible redox peaks with the formal potential of (E^o') of (-45±2) mV (vs. Ag/AgCl) at scan rate of 0.05 V/s. The heterogeneous electron transfer rate constant was evaluated to be 0.65 s^-1 in 50 mmol/L Na₂HPO₄-NaH₂PO₄ buffer solution at pH 7.0. The cathodic peak current of the modified electrode increased with increasing concentration of hydrogen peroxide(from 0.02 to 10 nmol/L) with a detection limit of 0.05 nmol/L. The apparent Michaelis-Menten constant (Kmapp) was 0.23 nmol/L.

Key words:

1. [1]
  1 LIU Sheng-Hua, WU Cheng-Tai. Chemisrty, 1998, 4: 1-51 LIU Sheng-Hua, WU Cheng-Tai. Chemisrty, 1998, 4: 1-5
2. [2]
  刘盛华, 吴成泰. 化学通报, 1998, 4: 1-5刘盛华, 吴成泰. 化学通报, 1998, 4: 1-5
3. [3]
  2 XING Jin-Juan, LIU Lin. Journal of Liaoning Institute of Technology-Natural Science Edition, 2009, 29(2): 125-1282 XING Jin-Juan, LIU Lin. Journal of Liaoning Institute of Technology-Natural Science Edition, 2009, 29(2): 125-128
4. [4]
  邢锦娟, 刘琳. 辽宁工业大学学报-自然科学版, 2009, 29(2): 125-128邢锦娟, 刘琳. 辽宁工业大学学报-自然科学版, 2009, 29(2): 125-128
5. [5]
  3 CAO Feng, REN Yong, HUA Wei-Yi, MA Kun-Fang, GUO Yan-Long. Chinese Journal of Organic Chemistry, 2002, 22(11): 827-8343 CAO Feng, REN Yong, HUA Wei-Yi, MA Kun-Fang, GUO Yan-Long. Chinese Journal of Organic Chemistry, 2002, 22(11): 827-834
6. [6]
  操锋, 任勇, 华维一, 马坤芳, 郭寅龙. 有机化学, 2002, 22(11): 827-834操锋, 任勇, 华维一, 马坤芳, 郭寅龙. 有机化学, 2002, 22(11): 827-834
7. [7]
  4 YUAN De-Qi, XIE Ru-Gang, ZHAO Hua-Ming. Chinese Joumal of Organic Chemistry, 1992, 12: 126-1384 YUAN De-Qi, XIE Ru-Gang, ZHAO Hua-Ming. Chinese Joumal of Organic Chemistry, 1992, 12: 126-138
8. [8]
  袁德其, 谢如刚, 赵华明. 有机化学, 1992, 12(2): 126-138袁德其, 谢如刚, 赵华明. 有机化学, 1992, 12(2): 126-138
9. [9]
  5 LI Yang, SUN Ji-Zhou, BIAN Chao, TONG Jian-Hua, XIA Shan-Hong. Chinese J. Anal. Chem., 2011, 39(11): 1621-16285 LI Yang, SUN Ji-Zhou, BIAN Chao, TONG Jian-Hua, XIA Shan-Hong. Chinese J. Anal. Chem., 2011, 39(11): 1621-1628
10. [10]
  李洋, 孙楫舟, 边超, 佟建华, 夏善红. 分析化学, 2011, 39(11): 1621-1628李洋, 孙楫舟, 边超, 佟建华, 夏善红. 分析化学, 2011, 39(11): 1621-1628
11. [11]
  6 HUANG Ying-Ping, YAN Ke-Mei, LUO Guang-Fu, LI Rui-Ping, CHEN Bai-Ling. Journal of China Three Gorges University-Natural Science Edition, 2001, 23(4): 368-3736 HUANG Ying-Ping, YAN Ke-Mei, LUO Guang-Fu, LI Rui-Ping, CHEN Bai-Ling. Journal of China Three Gorges University-Natural Science Edition, 2001, 23(4): 368-373
12. [12]
  黄应平, 颜克美, 罗光富, 李瑞萍, 陈百玲. 三峡大学学报-自然科学版, 2001, 23(4): 368-373黄应平, 颜克美, 罗光富, 李瑞萍, 陈百玲. 三峡大学学报-自然科学版, 2001, 23(4): 368-373
13. [13]
  7 CHEN Hai-Ming, LI Tong-Hua, CHEN Kai. Chinese J. Anal. Chem., 2002, 30(6): 654-6577 CHEN Hai-Ming, LI Tong-Hua, CHEN Kai. Chinese J. Anal. Chem., 2002, 30(6): 654-657
14. [14]
  陈海明, 李通化, 陈开. 分析化学, 2002, 30(6): 654-657陈海明, 李通化, 陈开. 分析化学, 2002, 30(6): 654-657
15. [15]
  8 Maggio G, Freni S, Cavallaro S. Power Sources, 1998, 74: 17-238 Maggio G, Freni S, Cavallaro S. Power Sources, 1998, 74: 17-23
16. [16]
  9 HAN Ai-Xia, REN Yan-Rong, HUANG Jing, LI Mei-Lan, WANG Huan-Lin, ZHANG Min, LI Wen-Lin. Journal of Guangzhou Chemical Industry, 2009, 37(5): 135-1379 HAN Ai-Xia, REN Yan-Rong, HUANG Jing, LI Mei-Lan, WANG Huan-Lin, ZHANG Min, LI Wen-Lin. Journal of Guangzhou Chemical Industry, 2009, 37(5): 135-137
17. [17]
  韩爱霞, 任彦蓉, 黄靖, 李梅兰, 汪焕林, 张敏, 李文林. 广州化工, 2009, 37(5): 135-137韩爱霞, 任彦蓉, 黄靖, 李梅兰, 汪焕林, 张敏, 李文林. 广州化工, 2009, 37(5): 135-137
18. [18]
  10 ZHENG Ning, GE Chen-Hao, GUO Zhong-Xian, LI Yuan-Zong, CHANG Wen-Bao, WANG Zong-Mu, LI Tie-Jin. Journal of Analytical Science, 2001, 17(4): 265-26910 ZHENG Ning, GE Chen-Hao, GUO Zhong-Xian, LI Yuan-Zong, CHANG Wen-Bao, WANG Zong-Mu, LI Tie-Jin. Journal of Analytical Science, 2001, 17(4): 265-269
19. [19]
  郑宁, 葛晨灏, 郭忠先, 李元宗, 常文保, 王宗睦, 李铁津. 分析科学学报, 2001, 17(4): 265-269郑宁, 葛晨灏, 郭忠先, 李元宗, 常文保, 王宗睦, 李铁津. 分析科学学报, 2001, 17(4): 265-269
20. [20]
  11 HUANG Ying-Ping, LUO Guang-Fu, LUO Yong-Jun, CAI Ru-Xiu. Chinese J. Anal. Chem., 2005, 33(5): 599-60411 HUANG Ying-Ping, LUO Guang-Fu, LUO Yong-Jun, CAI Ru-Xiu. Chinese J. Anal. Chem., 2005, 33(5): 599-604
21. [21]
  黄应平, 罗光富, 罗勇军, 蔡汝秀. 分析化学, 2005, 33(5): 599-604黄应平, 罗光富, 罗勇军, 蔡汝秀. 分析化学, 2005, 33(5): 599-604
22. [22]
  12 ZHENG QI, ZENG Yun-E, CAI Ru-Xiu. Journal of Analytical Science, 2005, 21(5): 563-56812 ZHENG QI, ZENG Yun-E, CAI Ru-Xiu. Journal of Analytical Science, 2005, 21(5): 563-568
23. [23]
  郑琦, 曾云鹗, 蔡汝秀. 分析科学学报, 2005, 21(5): 563-568郑琦, 曾云鹗, 蔡汝秀. 分析科学学报, 2005, 21(5): 563-568
24. [24]
  13 Rousseau C, Nielsen N, Bols M. Tetrahedron Letters, 2004, 45: 8709-871113 Rousseau C, Nielsen N, Bols M. Tetrahedron Letters, 2004, 45: 8709-8711
25. [25]
  14 Moosavi-Movahedi A A, Semsarha F, Heli H, Nazari K, Ghourchian H, Hong J, Hakimelahi G H, Saboury A A, Sefidbakht Y. Colloids and Surfaces A: Physicochem. Eng. Aspects, 2008, 320: 213-22114 Moosavi-Movahedi A A, Semsarha F, Heli H, Nazari K, Ghourchian H, Hong J, Hakimelahi G H, Saboury A A, Sefidbakht Y. Colloids and Surfaces A: Physicochem. Eng. Aspects, 2008, 320: 213-221
26. [26]
  15 Farivar F, Moosavi-Movahedi A A, Sefidbakht Y, Nazari K, Hong J, Sheibani N. Biochemical Engineering Journal, 2009, 49(1): 89-9415 Farivar F, Moosavi-Movahedi A A, Sefidbakht Y, Nazari K, Hong J, Sheibani N. Biochemical Engineering Journal, 2009, 49(1): 89-94
27. [27]
  16 Gharibi H, Moosavi-Movahedi Z, Javadian S, Nazari K, Moosavi-Movahedi A A. The Journal of Physical Chemistry B. 2011, 115: 4671-467916 Gharibi H, Moosavi-Movahedi Z, Javadian S, Nazari K, Moosavi-Movahedi A A. The Journal of Physical Chemistry B. 2011, 115: 4671-4679
28. [28]
  17 Hong J, Huang K, Wang W, Yang W-Y, Zhao Y-X, Xiao B-L, Moosavi-Movahedi Z, Ghourchian H, Sheibani N, Moosavi-Movahedi A A. Journal of the Iranian Chemical Society. 2012, In press. DOI: 10.1007/s13738-012-0078-317 Hong J, Huang K, Wang W, Yang W-Y, Zhao Y-X, Xiao B-L, Moosavi-Movahedi Z, Ghourchian H, Sheibani N, Moosavi-Movahedi A A. Journal of the Iranian Chemical Society. 2012, In press. DOI: 10.1007/s13738-012-0078-3
29. [29]
  18 YAO Shu-Qin, GUO Man-Dong. Physical Testing and Chemical Analysis Part B, 2004, 40(8): 488-49118 YAO Shu-Qin, GUO Man-Dong. Physical Testing and Chemical Analysis Part B, 2004, 40(8): 488-491
30. [30]
  姚淑琴, 郭满栋. 理化检验-化学分册, 2004, 40(8): 488-491姚淑琴, 郭满栋. 理化检验-化学分册, 2004, 40(8): 488-491
31. [31]
  19 Molaei Rad A, Ghourchian H, Moosavi-Movahedi A A, Hong J, Nazari K. Anal. Biochemi., 2007, 362: 38-4319 Molaei Rad A, Ghourchian H, Moosavi-Movahedi A A, Hong J, Nazari K. Anal. Biochemi., 2007, 362: 38-43

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

通讯作者: 洪军

English

Investigation of Self-assembled Nano-cluster Artificial Peroxidase by Voltammetry and Spectroscopy

Corresponding author: 洪军

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

光谱和电化学方法研究自组装纳米簇模拟过氧化物酶

通讯作者: 洪军

English

Investigation of Self-assembled Nano-cluster Artificial Peroxidase by Voltammetry and Spectroscopy

Corresponding author: 洪军

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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