Elaboration and characterization of spatially controlled assemblies of complementary polyphenol oxidase-alkaline phosphatase activities on electrodes

被引:40
作者
Mousty, C
Bergamasco, JL
Wessel, R
Perrot, H
Cosnier, S
机构
[1] Univ Grenoble 1, CNRS, UMR 5630, Lab Electrochim Organ & Photochim Redox, F-38041 Grenoble 9, France
[2] Univ Paris 06, CNRS, UPR 15, F-75252 Paris, France
关键词
D O I
10.1021/ac0100143
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The electrooxidation of a biotin pyrrole has allowed the formation of biotinylated polypyrrole films. Gravimetric measurements based on a quartz crystal microbalance demonstrate the efficient coupling of avidin, biotinylated polyphenol oxidase (PPO-B) and avidin-labeled alkaline phosphatase (AP-A) with the underlying biotinylated polymer film. The estimated mass increase corresponds to the anchoring of 1.6-1.8 equivalent layer of proteins. A step-by-step construction of bienzyme multilayers composed of PPO-B and AP-A was carried out on the electrode surface modified by the biotinylated polypyrrole film through avidin-biotin bridges. A spatially controlled distribution of the two enzymes was performed by the formation of one AP-A layer on 1, 5, and 10 PPO-B layers. The resulting bienzyme electrodes were applied to the determination of phenyl phosphate on the basis of amperometric detection of enzymically generated o-quinone at -0.2 V. Their analytical performances were analyzed in relation to the design of the enzyme architectures and in comparison with the amperometric behavior of the monoenzymatic electrodes (PPO-B electrode and AP-A electrode). It appears that at the 10-layer-PPO-B polypyrrole electrode only 4% of phenol is transformed, whereas 42-69% of phenyl phosphate is enzymatically consumed and detected at the AP-A polypyrrole electrode, depending on the enzyme activity, For the bienzymatic AP-A/PPO-B polypyrrole electrodes, the activity of each immobilized enzyme clearly affects the biosensor performance, the main limiting factor being the very low efficiency of PPO-B at pH 8.8.
引用
收藏
页码:2890 / 2897
页数:8
相关论文
共 36 条
  • [1] [Anonymous], 1987, Biosensors Fundamentals and Applications
  • [2] Enzyme sensors prepared by layer-by-layer deposition of enzymes on a platinum electrode through avidin-biotin interaction
    Anzai, J
    Kobayashi, Y
    Suzuki, Y
    Takeshita, H
    Chen, Q
    Osa, T
    Hoshi, T
    Du, XY
    [J]. SENSORS AND ACTUATORS B-CHEMICAL, 1998, 52 (1-2) : 3 - 9
  • [3] Anzai J., 1995, ANAL CHEM, V67, P811
  • [4] Automated amplified flow immunoassay for cocaine
    Bauer, CG
    Eremenko, AV
    Kühn, A
    Kürzinger, K
    Makower, A
    Scheller, FW
    [J]. ANALYTICAL CHEMISTRY, 1998, 70 (21) : 4624 - 4630
  • [5] Validation of antibody-based recognition by piezoelectric transducers through electroacoustic admittance analysis
    Bizet, K
    Gabrielli, C
    Perrot, H
    Therasse, J
    [J]. BIOSENSORS & BIOELECTRONICS, 1998, 13 (3-4) : 259 - 269
  • [6] A SUBSTRATE RECYCLING ASSAY FOR PHENOLIC-COMPOUNDS USING TYROSINASE AND NADH
    BROWN, RS
    MALE, KB
    LUONG, JHT
    [J]. ANALYTICAL BIOCHEMISTRY, 1994, 222 (01) : 131 - 139
  • [7] CEN Q, 1998, J ELECTROANALYSIS, V10, P94
  • [8] Poly(pyrrole-biotin): a new polymer for biomolecule grafting on electrode surfaces
    Cosnier, S
    Lepellec, A
    [J]. ELECTROCHIMICA ACTA, 1999, 44 (11) : 1833 - 1836
  • [9] An bienzyme electrode (alkaline phosphatase polyphenol oxidase) for the amperometric determination of phosphate
    Cosnier, S
    Gondran, C
    Watelet, JC
    De Giovani, W
    Furriel, RPM
    Leone, FA
    [J]. ANALYTICAL CHEMISTRY, 1998, 70 (18) : 3952 - 3956
  • [10] An original electroenzymatic system: Flavin reductase-riboflavin for the improvement of dehydrogenase-based biosensors. Application to the amperometric detection of lactate
    Cosnier, S
    Fontecave, M
    Innocent, C
    Niviere, V
    [J]. ELECTROANALYSIS, 1997, 9 (09) : 685 - 688