Abstract
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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Witteveen CF, Veenhuis M, Visser J (1992) Localization of glucose oxidase and catalase activities in Aspergillus niger. Appl Environ Microbiol 58:1190–1194
Wang J (2008) Electrochemical glucose biosensors. Chem Rev 108:814–825
Newman JD, Turner APF (2005) Home blood glucose biosensors: a commercial perspective. Biosens Bioelectron 20:2435–2453
Hanefeld U, Gardossi L, Magner E (2009) Understanding enzyme immobilisation. Chem Soc Rev 38:453–468
Lee SY, Choi JH, Xu Z (2003) Microbial cell-surface display. Trends Biotechnol 21:45–52
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 84:275–282
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 147:492–498
Liang B, Li L, Tang XL et al (2013) Microbial surface display of glucose dehydrogenase for amperometric glucose biosensor. Biosens Bioelectron 45:19–24
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 33:100–105
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 42:156–162
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 44:160–163
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 85:6107–6112
Boder ET, Wittrup KD (1997) Yeast surface display for screening combinatorial polypeptide libraries. Nat Biotechnol 15:553–557
Gouda MD, Singh SA, Rao AG et al (2003) Thermal inactivation of glucose oxidase. Mechanism and stabilization using additives. J Biol Chem 278:24324–24333
Bergmeyer HU (1974) Methods of enzymatic analysis, vol I, 2nd edn. Verlag Chemie, Academic, Weinheim, New York
Acknowledgements
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Wang, H., Lang, Q., Liang, B., Liu, A. (2015). Electrochemical Glucose Biosensor Based on Glucose Oxidase Displayed on Yeast Surface. In: Liu, B. (eds) Yeast Surface Display. Methods in Molecular Biology, vol 1319. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2748-7_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-2748-7_13
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2747-0
Online ISBN: 978-1-4939-2748-7
eBook Packages: Springer Protocols