CRCU

Chemical Research in Chinese Universities ›› 2011, Vol. 27 ›› Issue (5): 875-879.

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Hydrogen Peroxide Sensor Based on Horseradish Peroxidase Combined with CaCO3 Microspheres and Gold Nanoparticles

ZHENG Li1*, JIANG Feng-hua1, MA Guang-ran2, ZHUANG Qian-fen2 and LI Feng2*   

  1. 1. Marine Ecology Research Center, First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, P. R. China;
    2. Shandong Provincial Key Laboratory of Biochemical Analysis, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, P. R. China
  • Received:2010-12-06 Revised:2011-02-28 Online:2011-09-25 Published:2011-09-06
  • Contact: ZHENG Li;LI Feng E-mail:zhengli@fio.org.cn; lifeng@qust.edu.cn
  • Supported by:

    Supported by the National Natural Science Foundation of China(Nos.20775039, 20602009), the Natural Science Foundation of Shandong Province of China(No.ZR2009BM031), the Public Welfare Project of Marine Science Research, China(Nos. 200705011, 200805039), the Scientific Research Fund of the First Institute of Oceanography, the State Oceanic Administration of China(No.2010T04) and the Science and Technology Project of Shandong Company of China National Tobacco Corporation(No.KN172).

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Abstract: Direct electrochemistry and electrocatalysis of horseradish peroxidase(HRP) were achieved by entrapping the enzyme between CaCO3 microspheres and gold nanoparticles through forming sandwich configuration     (CaCO3-HRP-AuNPs). Polyanion, poly(styrene sulfonate)(PSS), was hybrid with CaCO3 microspheres to increase the surface negative charges for binding with HRP through electrostatic interaction. After the bioconjugate CaCO3 PSS-HRP was entrapped in chitosan based sol-gel(CS-GPTMS) film, HRP was encapsulated by in situ formation of an outer layer of AuNPs through electrochemical reduction of HAuCl4. The composite film containing AuNPs, CaCO3-PSS-HRP bioconjugates and CS-GPTMS can provide favorable microenvironment for HRP to perform direct electron transfer at glassy carbon electrode(GCE). HRP retained its bioelectrocatalytic activity and lead to sensitive and fast amperometric response for the determination of H2O2. H2O2 could be detected in a very wide linear range from 5.0×10–6 mol/L to 7.1×10–2 mol/L. The sandwich configuration of CaCO3-biomolecules-AuNPs could serve as a versatile platform for enzyme immobilization and biosensing.

Key words: Direct electrochemistry, Horseradish peroxidase(HRP), CaCO3 microsphere, Chitosan based sol-gel