Whole-cell method for phenol detection based on the color reaction of phenol with 4-aminoantipyrine catalyzed by CotA laccase on endospore surfaces.
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.
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Zhiming
Z
Zeng
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Longjian
L
Tian
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Zheng
Z
Li
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Lina
L
Jia
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Xinya
X
Zhang
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Miaomiao
M
Xia
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China; College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China. Electronic address: yongganghu@mail.hzau.edu.cn.
Yonggang
Y
Hu
D000675:Ampyrone / Q000494:pharmacology; D001407:Bacillus / Q000187:drug effects* / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001681:Biological Assay / Q000295:instrumentation*; D002384:Catalysis; D003124:Colorimetry / Q000295:instrumentation*; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000295:instrumentation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D042845:Laccase / Q000235:genetics / Q000378:metabolism*; D010636:Phenols / Q000032:analysis* / Q000494:pharmacology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014874:Water Pollutants, Chemical / Q000032:analysis
D001426:Bacterial Proteins; D010636:Phenols; D014874:Water Pollutants, Chemical; D000675:Ampyrone; C533003:CotA protein, Bacillus licheniformis; D042845:Laccase
Bacillus amyloliquefaciens; Coat protein A; Endospore; Phenol; Spectrophotometry