부산대학교 도서관

The electrochemical biosensors with enzymes have always been an attractive research topic in various sectors of biology due to their high specificity and reproducibility. In this study, we suggest a novel cellulosome scaffolding based multi-enzyme platform for the detection of isopropyl alcohol (IPA) using electrochemical biosensing assay. Three enzymes namely alcohol dehydrogenase (ADH), NADH oxidase (NOX), and peroxidase were isolated and successfully overexpressed in E.coli BL21 Rosetta respectively. The ability of enzymes to catalyze NAD+/NADH was confirmed using the absorption spectrum at 340㎚ wavelength. From the results, it was confirmed that the rate of oxidization of NADH by NOX and the rate of reduction of NAD+ by ADH were approximately 3 folds compared to control. In the synergistic reaction, comparative to the individual curves i.e. logistic for ADH and decaying for NOX, a straight line parallel to the baseline was obtained which confirmed that NAD+ was reduced to NADH and can be used by NOX for further oxidation simultaneously. Taken together, this study can be a progression in the area of electrochemical biosensors for the detection of IPA.