Electrochemical Glucose Biosensor Based on Glucose Oxidase Displayed on Yeast Surface

  • Hongwei Wang
  • Qiaolin Lang
  • Bo Liang
  • Aihua LiuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1319)


The conventional enzyme-based biosensor requires chemical or physical immobilization of purified enzymes on electrode surface, which often results in loss of enzyme activity and/or fractions immobilized over time. It is also costly. A major advantage of yeast surface display is that it enables the direct utilization of whole cell catalysts with eukaryote-produced proteins being displayed on the cell surface, providing an economic alternative to traditional production of purified enzymes. Herein, we describe the details of the display of glucose oxidase (GOx) on yeast cell surface and its application in the development of electrochemical glucose sensor. In order to achieve a direct electrochemistry of GOx, the entire cell catalyst (yeast-GOx) was immobilized together with multiwalled carbon nanotubes on the electrode, which allowed sensitive and selective glucose detection.

Key words

Yeast surface display Glucose oxidase Electrochemical biosensor Glucose 



This work was supported in part by National Natural Science Foundation of China (91227116, 31200982, 31200598, 21275152, and 21475144), and the Hundred-Talent-Project (No. KSCX2-YW-BR-7), the Chinese Academy of Sciences.


  1. 1.
    Witteveen CF, Veenhuis M, Visser J (1992) Localization of glucose oxidase and catalase activities in Aspergillus niger. Appl Environ Microbiol 58:1190–1194PubMedCentralPubMedGoogle Scholar
  2. 2.
    Wang J (2008) Electrochemical glucose biosensors. Chem Rev 108:814–825PubMedCrossRefGoogle Scholar
  3. 3.
    Newman JD, Turner APF (2005) Home blood glucose biosensors: a commercial perspective. Biosens Bioelectron 20:2435–2453PubMedCrossRefGoogle Scholar
  4. 4.
    Hanefeld U, Gardossi L, Magner E (2009) Understanding enzyme immobilisation. Chem Soc Rev 38:453–468PubMedCrossRefGoogle Scholar
  5. 5.
    Lee SY, Choi JH, Xu Z (2003) Microbial cell-surface display. Trends Biotechnol 21:45–52PubMedCrossRefGoogle Scholar
  6. 6.
    Liang B, Li L, Mascin M et al (2012) Construction of xylose dehydrogenase displayed on the surface of bacteria using ice nucleation protein for sensitive D-xylose detection. Anal Chem 84:275–282PubMedCrossRefGoogle Scholar
  7. 7.
    Liang B, Lang Q, Tang X et al (2013) Simultaneously improving stability and specificity of cell surface displayed glucose dehydrogenase mutants to construct whole-cell biocatalyst for glucose biosensor application. Bioresour Technol 147:492–498PubMedCrossRefGoogle Scholar
  8. 8.
    Liang B, Li L, Tang XL et al (2013) Microbial surface display of glucose dehydrogenase for amperometric glucose biosensor. Biosens Bioelectron 45:19–24PubMedCrossRefGoogle Scholar
  9. 9.
    Li L, Liang B, Shi JG et al (2012) A selective and sensitive D-xylose electrochemical biosensor based on xylose dehydrogenase displayed on the surface of bacteria and multi-walled carbon nanotubes modified electrode. Biosens Bioelectron 33:100–105PubMedCrossRefGoogle Scholar
  10. 10.
    Li L, Liang B, Li F et al (2013) Co-immobilization of glucose oxidase and xylose dehydrogenase displayed whole cell on multiwalled carbon nanotube nanocomposite films modified-electrode for simultaneous voltammetric detection of D-glucose and D-xylose. Biosens Bioelectron 42:156–162PubMedCrossRefGoogle Scholar
  11. 11.
    Xia L, Liang B, Li L et al (2013) Direct energy conversion from xylose using xylose dehydrogenase surface displayed bacteria based enzymatic biofuel cell. Biosens Bioelectron 44:160–163PubMedCrossRefGoogle Scholar
  12. 12.
    Wang H, Lang Q, Li L et al (2013) Yeast surface displaying glucose oxidase as whole-cell biocatalyst: construction, characterization, and its electrochemical glucose sensing application. Anal Chem 85:6107–6112PubMedCrossRefGoogle Scholar
  13. 13.
    Boder ET, Wittrup KD (1997) Yeast surface display for screening combinatorial polypeptide libraries. Nat Biotechnol 15:553–557PubMedCrossRefGoogle Scholar
  14. 14.
    Gouda MD, Singh SA, Rao AG et al (2003) Thermal inactivation of glucose oxidase. Mechanism and stabilization using additives. J Biol Chem 278:24324–24333PubMedCrossRefGoogle Scholar
  15. 15.
    Bergmeyer HU (1974) Methods of enzymatic analysis, vol I, 2nd edn. Verlag Chemie, Academic, Weinheim, New YorkGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Hongwei Wang
    • 1
    • 2
  • Qiaolin Lang
    • 1
  • Bo Liang
    • 1
  • Aihua Liu
    • 1
    Email author
  1. 1.Laboratory for Biosensing, Qingdao Institute of Bioenergy & Bioprocess Technology (QIBEBT) and Key Laboratory of Biofuels (QIBEBT)Chinese Academy of SciencesQingdaoChina
  2. 2.State Key Laboratory of Crop Biology, College of AgronomyShandong Agricultural UniversityTai’an, ShandongChina

Personalised recommendations