Covalent immobilization of horseradish peroxidase via click chemistry and its direct electrochemistry.
A simple and versatile approach for covalent immobilization of redox protein on solid surface via self-assembled technique and click chemistry is reported. The alkynyl-terminated monolayers are obtained by self-assembled technique, then, azido-horseradish peroxidase (azido-HRP) was covalent immobilized onto the formed monolayers by click reaction. The modified process is characterized by reflection absorption infrared spectroscopy (RAIR), surface-enhanced Raman scattering spectroscopy (SERS) and electrochemical methods. All the experimental results suggest that HRP is immobilized onto the electrode surface successfully without denaturation. Furthermore, the immobilized HRP shows electrocatalytic reduction for H(2)O(2), and the linear range is from 5.0 to 700 μM. The heterogeneous electron transfer rate constant k(s) is 1.11 s(-1) and the apparent Michaelis-Menten constant is calculated to be 0.196 mM.
Department of Chemistry, East China Normal University, Shanghai, China.
Qin
Q
Ran
Ru
R
Peng
Cong
C
Liang
Siqiu
S
Ye
Yuezhong
Y
Xian
Wenjing
W
Zhang
Litong
L
Jin
D015374:Biosensing Techniques / Q000639:trends; D004563:Electrochemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006735:Horseradish Peroxidase / Q000737:chemistry*; D019265:Spectroscopy, Near-Infrared
D004800:Enzymes, Immobilized; D006735:Horseradish Peroxidase