Electrochemical characterization of enzymatic activity of yeast cells entrapped in a poly(dimethylsiloxane) microwell on the basis of limited diffusion system.
A highly sensitive and quantitative analysis was performed using a poly(dimethylsiloxane) (PDMS) microwell array in a scanning electrochemical microscopy setup. A microelectrode with a relatively large seal radius was used to cover the top of the cylindrical PDMS microwell (96 pL). The voltammogram for 4 mM ferrocyanide resulted in a charge value of 38 nC, suggesting that almost 100% of the reductant in the microwell was converted to the oxidation current. When genetically modified yeast cells were entrapped in the microwell, the accumulation of p-aminophenol (PAP) produced by expressing beta-galactosidase (betaGAL) was successfully observed.
Shiku
Hitoshi
H
Graduate School of Environmental Studies, Tohoku University, 6-6-11, Aramaki-Aoba, Sendai 980-8579, Japan. shiku@bioinfo.che.tohoku.ac.jp
Goto
Shun
S
Jung
Sungbong
S
Nagamine
Kuniaki
K
Koide
Masahiro
M
Itayama
Tomosato
T
Yasukawa
Tomoyuki
T
Matsue
Tomokazu
T
D004058:Diffusion; D004129:Dimethylpolysiloxanes; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D008839:Microelectrodes; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D001616:beta-Galactosidase / Q000032:analysis*
D004129:Dimethylpolysiloxanes; D001616:beta-Galactosidase