Escherichia coli (E. coli) is one of the most ubiquitous foodborne pathogens that can cause infections and threaten human health. Herein, a colorimetric method for sensitive detection of E. coli was established by using enzyme-nanozyme cascade reaction for signal amplification. Gold nanoparticles (AuNPs) are well-known nanozymes due to their high peroxidase-like activity. When the dense cetyltrimethylammonium bromide (CTAB) membrane on the surfaces of AuNPs kept the substrate away from AuNPs, the peroxidase activity of AuNPs was inhibited. However, the CTAB membrane could be disrupted by Ag+, resulting in enhanced peroxidase activity of AuNPs. When E. coli was present, the enzyme-nanozyme cascade reaction was initiated. The substrate p-aminophenyl β-D-galactopyranoside (PAPG) was hydrolyzed to the reductive p-aminophenol (PAP) by beta-galactosidase (β-gal) in E. coli, reducing Ag+ to Ag. Consequently, CTAB-AuNPs remained weak peroxidase activity and could not catalyze the H2O2-mediated oxidation of TMB. As the amount of E. coli increased, the absorbance of TMB decreased along with a color change from deep blue to pink. The absorbance intensity displayed a linear dependence on E. coli from 1.0 × 102 to 1.0 × 109 CFU mL-1. Therefore, the proposed method holds good prospects in foodborne pathogenic bacteria detection.
Keywords: Bacteria; CTAB-AuNPs; Cascade reaction; Peroxidase activity.
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