Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors

Sung Gil Hong, Jae Hyun Kim, Ryang Eun Kim, Seok Joon Kwon, Dae Woo Kim, Hee Tae Jung, Jonathan S. Dordick, Jungbae Kim

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Glucose oxidase (GOx) was immobilized onto graphene oxide (GRO) via three different preparation methods: enzyme adsorption (EA), enzyme adsorption and crosslinking (EAC), and enzyme adsorption, precipitation and crosslinking (EAPC). EAPC formulations, prepared via enzyme precipitation with 60% ammonium sulfate, showed 1,980 and 1,630 times higher activity per weight of GRO than those of EA and EAC formulations, respectively. After 59 days at room temperature, EAPC maintained 88% of initial activity, while EA and EAC retained 42 and 45% of their initial activities, respectively. These results indicate that the steps of precipitation and crosslinking in the EAPC formulation are critical to achieve high enzyme loading and stability of EAPC. EA, EAC and EAPC were used to prepare enzyme electrodes for use as glucose biosensors. Optimized EAPC electrode showed 93- and 25-fold higher sensitivity than EA and EAC, respectively. To further increase the sensitivity of EAPC electrode, multi-walled carbon nanotubes (MWCNTs) were mixed with EAPC for the preparation of enzyme electrode. Surprisingly, the EAPC electrode with additional 99.5 wt% MWCNTs showed 7,800-fold higher sensitivity than the EAPC electrode without MWCNT addition. Immobilization and stabilization of enzymes on GRO via the EAPC approach can be used for the development of highly sensitive biosensors as well as to achieve high enzyme loading and stability.

Original languageEnglish
Pages (from-to)573-579
Number of pages7
JournalBiotechnology and Bioprocess Engineering
Volume21
Issue number4
DOIs
Publication statusPublished - 2016 Aug 1

Fingerprint

Glucose Oxidase
Glucose oxidase
Graphite
Biosensing Techniques
Biosensors
Immobilization
Oxides
Graphene
Adsorption
Enzymes
Crosslinking
Carbon Nanotubes
Electrodes
Enzyme electrodes
Enzyme Stability
Carbon nanotubes

Keywords

  • biosensors
  • carbon nanotubes
  • enzyme adsorption/precipitation/crosslinking
  • glucose oxidase
  • graphene oxide

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Biomedical Engineering

Cite this

Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors. / Hong, Sung Gil; Kim, Jae Hyun; Kim, Ryang Eun; Kwon, Seok Joon; Kim, Dae Woo; Jung, Hee Tae; Dordick, Jonathan S.; Kim, Jungbae.

In: Biotechnology and Bioprocess Engineering, Vol. 21, No. 4, 01.08.2016, p. 573-579.

Research output: Contribution to journalArticle

Hong, Sung Gil ; Kim, Jae Hyun ; Kim, Ryang Eun ; Kwon, Seok Joon ; Kim, Dae Woo ; Jung, Hee Tae ; Dordick, Jonathan S. ; Kim, Jungbae. / Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors. In: Biotechnology and Bioprocess Engineering. 2016 ; Vol. 21, No. 4. pp. 573-579.
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