TY - JOUR
T1 - Electrochemical tuning of alcohol oxidase and dehydrogenase catalysis via biosensing towards butanol-1 detection in fermentation media
AU - Semenova, Daria
AU - Pinto, Tiago
AU - Koch, Marcus
AU - Gernaey, Krist V.
AU - Junicke, Helena
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - Alcohol dehydrogenase
KW - Alcohol oxidase
KW - Amperometric biosensors
KW - Biobutanol production
KW - Butanol determination
KW - Pd-NPs
U2 - 10.1016/j.bios.2020.112702
DO - 10.1016/j.bios.2020.112702
M3 - Journal article
C2 - 33045667
AN - SCOPUS:85092405052
SN - 0956-5663
VL - 170
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 112702
ER -