Direct-Electron-Transfer Bio-Nanoink with Single-Walled Carbon Nanotube and Aspergillus terreus var. aureus Flavin Adenine Dinucleotide Glucose Dehydrogenase

Atsunori Hiratsuka, Hisanori Iwasa, Hirotaka Uzawa, Atsuya Suzuki, Hitoshi Muguruma

Research output: Contribution to journalArticle

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Abstract

This pioneering study contains the fabrication of a novel concept of direct-electron-transfer (DET) bio-nanoink for a mediator-free and oxygen-insensitive glucose sensing. The DET bio-nanoink is composed of glycan chain-rich flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase (GDH), single-walled carbon nanotubes (SWCNTs), and surfactant sodium cholate in aqueous solution. We discovered FAD-GDH genes in various fungi and characterized the enzymatic properties of the recombinant enzymes produced by Pichia pastoris as a host. Glycan-chain-rich Aspergillus terreus var. aureus FAD-GDH is screened and is the most suitable for the bio-nanoink compared to conventional FAD-GDHs from Aspergillus species in terms of chemical stability and activity. The amperometric biosensor is fabricated with piezoelectric inkjet printing, where the small fraction (several pL) of the bio-nanoink can be erupted without clogging of the nozzle. The biosensor with the bio-nanoink showed a large (1 mA cm -2 at +0.6 V 48 mM glucose) and a glucose-concentration-dependent current, indicating that the DET between FAD-GDH and SWCNTs in the bio-nanoink is accomplished. The DET is supported by the observation of the atomic force and transmission microscopies, where debundled SWCNTs are connected to FAD-GDH molecules. The mediator-free and oxygen-insensitive biosensor fabricated by the DET bio-nanoink revealed a high sensitivity of 70 μA mM -1 cm -2 , a broad linear dynamic range of 0.0025-3.2 mM, and selectivity toward an interferent, and a low detection limit of 1.1 μm, which are superior to those of any other glucose biosensor.

Original languageEnglish
Pages (from-to)5776-5783
Number of pages8
JournalACS Omega
Volume4
Issue number3
DOIs
Publication statusPublished - 2019 Mar 25

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Glucose 1-Dehydrogenase
Flavin-Adenine Dinucleotide
Aspergillus
Single-walled carbon nanotubes (SWCN)
Glucose
Biosensors
Electrons
Polysaccharides
Sodium Cholate
Oxygen
Chemical stability
Fungi
Surface-Active Agents
Oxidoreductases
Printing
Nozzles
Microscopic examination
Genes
Fabrication
Molecules

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

Cite this

Direct-Electron-Transfer Bio-Nanoink with Single-Walled Carbon Nanotube and Aspergillus terreus var. aureus Flavin Adenine Dinucleotide Glucose Dehydrogenase. / Hiratsuka, Atsunori; Iwasa, Hisanori; Uzawa, Hirotaka; Suzuki, Atsuya; Muguruma, Hitoshi.

In: ACS Omega, Vol. 4, No. 3, 25.03.2019, p. 5776-5783.

Research output: Contribution to journalArticle

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