Electrochemical tuning of alcohol oxidase and dehydrogenase catalysis via biosensing towards butanol-1 detection in fermentation media

Daria Semenova*, Tiago Pinto, Marcus Koch, Krist V. Gernaey, Helena Junicke

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

A novel approach for electrochemical tuning of alcohol oxidase (AOx) and alcohol dehydrogenase (ADH) biocatalysis towards butanol-1 oxidation by incorporating enzymes in various designs of amperometric biosensors is presented. The biosensors were developed by using commercial graphene oxide-based screen-printed electrodes and varying enzyme producing strains, encapsulation approaches (layer-by-layer (LbL) or one-step electrodeposition (EcD)), layers composition and structure, operating conditions (applied potential values) and introducing mediators (Meldola Blue and Prussian Blue) or Pd-nanoparticles (Pd-NPs). Simultaneous analysis/screening of multiple crucial system parameters during the enzyme engineering process allowed to identify within a period of one month that four out of twelve proposed designs demonstrated a good signal reproducibility and linear response (up to 14.6 mM of butanol) under very low applied potentials (from −0.02 to −0.32 V). Their mechanical stability was thoroughly investigated by multi-analytical techniques prior to butanol determination in cell-free samples from an anaerobic butanol fermentation. The EcD-based biosensor that incorporates ADH, NAD+, Pd-NPs and Nafion showed no loss of enzyme activity after preparation and demonstrated capabilities towards low potential (−0.12 V) detection of butanol-1 in fermentation medium (4 mM) containing multiple electroactive species with almost 15 times enhanced sensitivity (0.2282 μA/mM ± 0.05) when compared to the LbL design. Furthermore, the ADH-Nafion bonding for the S. cerevisiae strain was confirmed to be 3 times higher than for E. coli.

Original languageEnglish
Article number112702
JournalBiosensors and Bioelectronics
Volume170
Number of pages8
ISSN0956-5663
DOIs
Publication statusPublished - 2020

Keywords

  • Alcohol dehydrogenase
  • Alcohol oxidase
  • Amperometric biosensors
  • Biobutanol production
  • Butanol determination
  • Pd-NPs

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