Supplement 1
Properties of (a) the soil sample (S3) and (b) the polymer industrial wastewater used in this study
a
Properties
pH
Organic matter (%)
Nitrogen (%)
Phosphorus (mg/kg)
Potassium (mg/kg)
Organic carbon (%)
C:N ratio
Moisture (%)
b
Properties
Color
pH
Electrical conductivity (EC, dS.m-1)
Salinity (ppt)
Total dissolved solid (TDS, mg l-1)
COD
Soil S3
6.82
5.34
2.45
58
640
37.4
15.3
23.9
Value
Colorless
9.24-9.69
57.1-75.3
38.2-51.2
45,507-55,998
680-3,170
Analytical method
Observation
pH meter
Electrical conductivity meter
Electrical conductivity meter
EPA 160.1
Potassium dichromate digestion
Supplement 2
Standard curves for the determination of 4CA concentration in environmental samples generated by (a) the
DH5/pPXGFP-pTODST bioreporter, and (b) HPLC. The standard curves were prepared as described in
Materials and methods. The bioreporter standard curve was generated within a linear concentration range of
4CA, from the average value of six replicates and the error bars represent the standard deviations. The linear
regression equation is y = 6.420x+3.07 (r2 = 0.991); where y is fluorescence induction ratio and x is 4CA
concentration. The HPLC standard curve was generated from three replicates and the error bars represent the
standard deviations. The linear regression equation is y = 7e6x – 212,366 (r2 = 0.996); where y is the arbitrary
Induction ratio
12
HPLC peak area (AU)
unit of HPLC peak area and x is 4CA concentration.
a
10
8
6
4
2
0
0.0
0.2
0.4
0.6
0.8
4CA concentration (mM)
1.0
b
8.E+06
6.E+06
4.E+06
2.E+06
0.E+00
0
0.4
0.8
1.2
4CA concentration (mM)
Supplement 3
A time-course fluorescence induction in the DH5/pPXGFP-pTODST in different culture media. Cells were resuspended in (a, d) LB, (b, e) M9G and (c, f) TB medium and incubated alone (control,) or with 1 mM of
either: toluene (), and 4CA () as the inducers. At intervals, the cell growth (opened symbol, dashed line) and
relative fluorescence unit (RFU) were evaluated and are shown as the (a, b, c) RFU or (d, e, f) the induction
ratio. The data points represent the means of at least three independent repeats, and the error bars represent the
300
a
b
c
2.0
200
1.5
1.0
Induction ratio
100
0.5
0
10
0.0
d
8
e
f
6
4
2
0
0
6
12
18
24 0
6
12
18
Incubation time (h)
24 0
6
12
18
24
Cell biomass (OD600)
Relative fluorescence
unit (RFU)
standard errors of the mean.
Journal of Industrial Microbiology & Biotechnology
Title:
Construction and application of an Escherichia coli bioreporter for aniline and
chloroaniline detection
Authors:
Alisa S. Vangnaia,b*, Naoya Kataokac, Suwat Soonglerdsongphad, Chatvalee Kalambahetid,
Takahisa Tajimac, and Junichi Katoc
a
Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330,
Thailand
b
National Center of Excellence for Environmental and Hazardous Waste Management (NCEEHWM), Chulalongkorn University, Bangkok 10330 Thailand
c
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter,
Hiroshima University, Hiroshima 739-8530, Japan
d
Environmental Research and Management Department, PTT Research and Technology
Institute, Wangnoi, Ayutthaya 13170, Thailand
*Corresponding author
Correspondence:
Alisa S. Vangnai
Department of Biochemistry, Faculty of Science,
Chulalongkorn University, 254 Phayathai Road, Bangkok 10330, Thailand.
Tel: +662 218 5430; +662 218 5418;
E-mail: alisa.v@chula.ac.th, avangnai@yahoo.com