TY - JOUR
T1 - A new method for determining activity of CMEPA hydrolase
AU - Wang, Fei
AU - Li, Zhou Kun
AU - Dong, Wei Liang
AU - Cui, Zhong Li
N1 - Publisher Copyright:
© 2016, China Environmental Science Press. All rights reserved.
PY - 2016/7/25
Y1 - 2016/7/25
N2 - A new method for rapid determination of the activity of 2'-methyl-6'-ethyl-2-chloroacetanilide(CMEPA) hydrolase in Delftia acidovorans T3-6 was established. CMEPA hydrolase can hydrolyze CMEPA into 2-methyl-6-ethyl-aniline (MEA), which, in turn, reacts with 4-aminoantipyrene in 20 mmol·L-1 Tris-HCl (pH 7.0) buffer to produce a compound, purple in color, with absorbancy at 535 nm(y) positively related to the concentration of MEA(x) in the range of 10-50 µmol·L-1. The linear equation goes as y=0.014 6x-0.005 4, R2=0.9988. The compounds of CMEPA, SDS, organic reagent such as methanol and 1 mmol·L-1 metal ions have no significant effect on chromogenic reaction, whereas temperature and pH does. Compared with the dichloromethane extraction-UV spectrophotometry method, the 4-aminoantipyrene colorimetric method does not vary much in determining enzymatic activity of T3-6 crude enzyme. This method is also applicable to the determination of other derivatives of aniline and phenol.
AB - A new method for rapid determination of the activity of 2'-methyl-6'-ethyl-2-chloroacetanilide(CMEPA) hydrolase in Delftia acidovorans T3-6 was established. CMEPA hydrolase can hydrolyze CMEPA into 2-methyl-6-ethyl-aniline (MEA), which, in turn, reacts with 4-aminoantipyrene in 20 mmol·L-1 Tris-HCl (pH 7.0) buffer to produce a compound, purple in color, with absorbancy at 535 nm(y) positively related to the concentration of MEA(x) in the range of 10-50 µmol·L-1. The linear equation goes as y=0.014 6x-0.005 4, R2=0.9988. The compounds of CMEPA, SDS, organic reagent such as methanol and 1 mmol·L-1 metal ions have no significant effect on chromogenic reaction, whereas temperature and pH does. Compared with the dichloromethane extraction-UV spectrophotometry method, the 4-aminoantipyrene colorimetric method does not vary much in determining enzymatic activity of T3-6 crude enzyme. This method is also applicable to the determination of other derivatives of aniline and phenol.
KW - 2-methyl-6-ethyl-aniline (MEA)
KW - 4-aminoantipyrinechromogenic method
KW - Spectrophotometry delftia acidovorans T3-6
UR - http://www.scopus.com/inward/record.url?scp=85042509945&partnerID=8YFLogxK
U2 - 10.11934/j.issn.1673-4831.2016.04.026
DO - 10.11934/j.issn.1673-4831.2016.04.026
M3 - 文章
AN - SCOPUS:85042509945
SN - 1673-4831
VL - 32
SP - 682
EP - 686
JO - Journal of Ecology and Rural Environment
JF - Journal of Ecology and Rural Environment
IS - 4
M1 - 1673-4831(2016)04-0682-05
ER -