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標題: Voltammetric measurement of Escherichia coli concentration through p-APG hydrolysis by endogenous β-galactosidase
作者: Yu-Jui Fan Ph.D. 
Yu-Chen Hsu master degree 
Bing-Chen Gu master degree 
Chia-Che Wu Ph.D. 
This paper reports a facile electrochemical detection method for Escherichia coli (E. coli) that does not use DNA amplification or immunoassay. The detection principle is based on the activity of the β-galactosidase (β-gal) endogenous enzyme, which hydrolyzes p-aminophenyl-β-d-galactopyranoside (p-APG) into p-aminophenol. After E. coli consumes p-APG within 30 min, the remaining p-APG is oxidized on a gold electrode using cyclic voltammetry and square wave voltammetry. The β-gal expression level is increased through treatment with a β-gal expression inducer (isopropyl-β-d-thiogalactopyranoside), and the hydrolysis reaction of p-APG is facilitated through permeabilization treatment with sodium dodecyl sulfate. The calibration curve for E. coli has a working range of 102–104 colony-forming units per mL in nutrient broth buffer. The total assay time is less than 100 min. The successful application of this approach indicates the possibility of rapid detection.
Appears in Collections:機械工程學系所

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