16 1 Vol 16 No 1 2010 2 ELECTROCHEMISTRY Feb 2010 1006-3471 2010 01-0065-05 - L- /GC * 200234 L- 1- - 3- L- L- O646 A Chloroperoxidase CPO 1 1 CPO Sigma L- Sigma 50 mmol /L ph = 9 0 10 1 mmol /L 1- - 3- P-450 2-4 1-3-dimethylamino propyl -3-eth- 5 ylcarbo-diimide hydrochloride 99% EDC 6-7 8 CPO Heal Force CPO 50 mmol /L CPO PBS KH 2 PO 4 -Na 2 HPO 4 ph 0 1 mol /L KCl 1 2 L- CHI 650 9 10 11 Rusling GC = 3 mm L- Poly-L-lysine PLL 12-15 L- GC 1- - 3- CPO PLL / 1 3 GC GC 6# 0 3 μm 1 2009-07-25 2009-08-25 * Tel 86-21 64321648 E-mail xqwu@ shnu edu cn 20973114 20775049 07QA14044
66 2010 0 05 μm α-al 2 O 3 CPO Fe Ⅲ / Fe Ⅱ 16-0 220 V GC 10 mmol /L ph = 9 0 L- - 1 0 ~ 2 5 V CPO PLL /GC 100 mv /s PLL CPO PLL PLL /GC 1 μl 2 3 10-4 mmol /L CPO 4 μl 24 mmol /L EDC PLL /GC CPO- PLL /GC 2 2 1 L- - 0 242 V Ep 22 mv 100 mv /s 1 L- L- 8 L- 2 GC a PLL /GC b CPO-PLL /GC c PLL 8 Fig 2 Cyclic voltammograms of bare GC a PLL /GC b 8 and CPO-PLL /GC electrode c in N 2 -saturated 50 mmol /L PBS ph = 2 75 containing 0 1 mol /L KCl scan rate is 100 mv /s 1 Fig 1 L- 0 9992 Cyclic voltammograms of electropolymerization for 10 mmol /L L-lysine ph = 9 0 at GC electrode scan rate is 100 mv /s 3 CPO PLL /GC 1 0 1 ~ 1 5 V /s 17 Ipa μa = 0 015v mv /s + 0 43 R 1 = 0 9984 Ipc μa = - 0 022v mv / s - 0 31 R 2 = 2 3 ph 4 CPO-PLL/GC ph 3 0 ~ 8 0 2 2 PLL /GC CPO ph ph 2 ph 2 75 CPO Fe Ⅲ /Fe Ⅱ GC a PLL / GC b E 0 PLL /GC CPO c - 0 23 V ph E 0 V = - 0 0322
1 - L- /GC 67 3 CPO-PLL /GC A B Fig 3 Cyclic voltammograms of CPO-PLL /GC electrode in a N 2 -saturated 50 mmol /L PBS ph = 2 75 containing 0 1 mol /L KCl scan rate a ~ g /V s - 1 0 1 0 3 0 5 0 8 1 1 2 1 5 A and the peak current versus scan rate B 4 Fig 4 CPO-PLL /GC ph PBS A E 0 ~ ph B Cyclic voltammograms of CPO-PLL /GC in different ph a ~ f 3 0 4 0 5 0 6 0 7 0 8 0 PBS containing 0 1 mol /L KCl A and the of formal potential E 0 versus ph B scan rate 100 mv /s ph - 0 1272 R = 0 9994 32 mv /ph 57 6 mv /ph 20 2 4 CPO-PLL /GC 5 CPO-PLL /GC 100 mv /s 100 CPO-PLL /GC 40 d 5 CPO-PLL /GC N CPO 2 0 1 mol /L KCl 50 mmol /L PBS 2 5 CPO-PLL /GC 100 Fig 5 The successive 100 cyclic voltammograms of CPO- PLL /GC electrode in a N 2 -saturated 50 mmol /L 6 GC a PLL /GC b CPO-PLL /GC PBS ph = 2 75 containing 0 1 mol /L KCl c scan rate 100 mv /s
68 2010 GC - 0 82 V al C fumago chloroperoxidase is also a dehaloperoxidase oxidative dehalogenation of halophenols J J Am PLL /GC - 0 55 V Chem Soc 2006 128 4 1036-1037 CPO - 0 2 V 4 John H Dawson Masanori Sono Cytochrome P-450 and GC PLL /GC chloroperoxidase thiolate-ligated heme enzymes Spectroscopic determination of their active site structures and 620 mv 350 mv mechanistic implications of thiolate ligation J Chem CPO Rev 1987 87 5 1255-1276 6 Fig 6 chloroperoxidase at GC electrode modified with SWNTs GC a PLL /GC b CPO-PLL /GC c in Chinese J Electrochemistry 2008 14 50 mmol /L PBS ph = 2 75 Cyclic voltammograms of the bare GC a PLL /GC b and CPO-PLL /GC c electrodes in O 2 -saturated 50 mmol /L PBS ph = 2 75 scan rate 100 mv /s 3 PLL CPO xin Cyclic voltammetric determination of dopamine in drug using poly L-lysine modified electrode J Chinese Journal of Analytical Chemistry CPO CPO-PLL /GC 2006 34 5 668-670 10 Zhao Yan-xia Ma Xin-ying Wang CPO-PLL /GC Qi et al Electrocatalytic oxidation to norepinephrine at a poly L-lysine modified electrode J Chinese Journal of Analysis Laboratory References 1 Cao Shu-yu Liu Jun-hong Biotechnology progress on applications of peroxidase J Journal of Chemical Industry & Engineering 2003 24 5 28-32 2 Lowell P Hager David R Morris Frederick S Brown et al Chloroperoxidase Ⅱ Utilization of halogen anions J Biological Chem 1966 241 8 1769-1777 3 Robert L Osborne Gregory M Raner Lowell P Hager et 5 Lu Zhong-qing Hou Wen-jing Wu Xia-qin et al Electrochemistry of self-assembled ultra-thin films composed of chloroperoxidase and polyions J Chem Lett 2007 36 4 564-565 6 Zhang Hong-xia Wu Xia-qin Hou Wen-jing 瞡 et al Catalytic chlorination of MCD using chloroperoxidase coated single-wall carbon nanotubes films on GC electrodes J Chemical J of Chinese University 2008 29 9 1863-1865 7 Zhang Hong-xia Wu Xia-qin Hou Wen-jing 瞡 et al Direct electrochemistry of 4 383-387 8 Yang Liu Wu Xia-qin Wang Rong et al Ionic liquid modified GC electrode for the direct electrochemistry of chloroperoxidase J Chinese Chemical Letters 2008 19 12 1483-1486 9 Sun Deng-ming Ma Wei Zhang Zhen - 2005 24 3 71-74 11 Wang Xiu-wen Yu Fen Zhao Zhijie et al Electrochemical behavior of acetaminophen at poly L-lysine-modified electrode and its detection J Chemistry & Bioengineering 2008 25 1 68-72 12 Peterson M Guto Challa V Kumar James F Rusling Thermostable peroxidase-polylysine films for biocatalysis at 90 J J Phys Chem B 2007 111 30
1 - L- /GC 69 9125-9131 13 Abhay Vaze Michael Parizo James F Rusling Enhanc- 13 ed rates of electrolytic styrene epoxidation catalyzed by cross-linked myoglobin-poly L-lysine films in bicontinuous microemulsions J Langmuir 2004 20 25 10943-10948 14 Peterson M Guto James F Rusling Enzyme-like kinetics of ferryloxy myoglobin formation in films on electrodes in microemulsions J J Phys Chem B 2005 109 51 24457-24464 15 Abhay Vaze James F Rusling Microemulsion-controll- 15 ed reaction sites in biocatalytic films for electrochemical reduction of vicinal dibromides J Langmuir 2006 22 25 10788-10795 16 Zhang Zhe Salem Chouchane Richard S Magliozzo et al Direct voltammetry and catalysis with mycobacterium tuberculosis catalase-peroxidase peroxidases and catalase in lipid films J Anal Chem 2002 74 1 163-170 17 Alan P Brown Fred C Anson Electron transfer kinetics with both reactant and product attached to the electrode surface J Journal of Electroanalytical Chemistry 1978 92 2 133-145 The Electrochemical Characteristics of CPO-PLL / GC Electrode ZHAO Wen WU Xia-qin * LU Zhong-qing HOU Wen-jing LI He-xing Life and Environment Science College Shanghai Normal University Shanghai 200234 China Abstract Poly-L-lysine PLL was electropolymerized onto the surface of glassy carbon GC electrode 1-3-dimethylamino propyl -3-ethylcarbodiimide hydrochloride EDC was used as a Cross-linking agent to immobilized Chloroperoxidase CPO onto the PLL /GC electrode A pair of well-defined redox peaks was observed at CPO-PLL /GC electrode which clearly suggested that PLL can promote the direct electron transfer between the CPO and the GC electrodes The cyclic voltammetry results also showed that the electron transfer at CPO-PLL / GC is a surface adsorption-controlled quasi-reversible process which is accompanied with proton transfer The CPO-PLL / GC electrode exhibited a good electrochemical stability and excellently electrocatalytic activity for electrochemical reduction of O 2 Key words chloroperoxidase poly-l-lysine electropolymerization direct electrochemistry electrocatalysis modified electrode