Whole-cell method for phenol detection based on the color reaction of phenol with 4-aminoantipyrine catalyzed by CotA laccase on endospore surfaces

Biosens Bioelectron. 2015 Jul 15:69:162-6. doi: 10.1016/j.bios.2015.02.032. Epub 2015 Feb 20.

Abstract

A green method for phenol spectrophotometric determination was developed based on the color reaction of phenol with 4-aminoantipyrine catalyzed by addition of Bacillus amyloliquefaciens endospores in the presence of O2. The catalytic activity of the endospores may be attributed to the presence of coat protein A on the cell surfaces. This deduction was confirmed by cotA gene knock-out from B. amyloliquefaciens using the homologous double-exchange method. Under optimal conditions, linear responses were obtained over phenol concentrations ranging from 5.0×10(-5)gL(-1) to 1.0×10(-2)gL(-1) (r=0.9984) with a detection limit of 2.1×10(-5)gL(-1) (3σ). Repeatability measurements of 1.0mgL(-1) phenol provided reproducible results with a relative standard deviation of 5.3% (n=11). Standard addition tests indicated recoveries ranging from 92.78% to 107.60%. The proposed whole-cell method was successfully used to detect total phenol in synthetic samples. Results confirmed the potential use of the developed method in practical applications.

Keywords: Bacillus amyloliquefaciens; Coat protein A; Endospore; Phenol; Spectrophotometry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ampyrone / pharmacology
  • Bacillus / drug effects*
  • Bacillus / physiology
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Biological Assay / instrumentation*
  • Catalysis
  • Colorimetry / instrumentation*
  • Dose-Response Relationship, Drug
  • Environmental Monitoring / instrumentation
  • Equipment Design
  • Equipment Failure Analysis
  • Laccase / genetics
  • Laccase / metabolism*
  • Phenols / analysis*
  • Phenols / pharmacology*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Water Pollutants, Chemical / analysis

Substances

  • Bacterial Proteins
  • Phenols
  • Water Pollutants, Chemical
  • Ampyrone
  • CotA protein, Bacillus licheniformis
  • Laccase