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Purification and characterization of a novel cypermethrin‐hydrolyzing esterase from Bacillus licheniformis B‐1
Author(s) -
Zhang Mengmei,
Lai Wen,
Zhu Yuanting,
Chen Shujuan,
Zhou Kang,
Ao Xiaolin,
He Li,
Yang Yong,
Zou Likou,
Liu Aiping,
Yao Kai,
Liu Shuliang
Publication year - 2021
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/1750-3841.15662
Subject(s) - esterase , chemistry , bacillus licheniformis , enzyme , chromatography , ammonium sulfate precipitation , enzyme assay , cypermethrin , molecular mass , size exclusion chromatography , biochemistry , bacillus subtilis , bacteria , biology , pesticide , agronomy , genetics
Cypermethrin (CY) is a synthetic pyrethroid widely used to control insect pests and it elicits a toxic effect on the human body. In this study, Bacillus licheniformis B‐1 isolated from tea garden soil was used to degrade CY effectively. A specific enzyme was mainly localized in the extracellular compartments of B‐1. This enzyme was identified as an esterase that could be produced without CY. The enzyme was purified 23.03‐fold to apparent homogeneity with 8.38% overall recovery by ammonium sulfate precipitation, anion exchange chromatography, and gel filtration chromatography. The molecular mass of the CY‐degrading enzyme was 66.4 kDa, and its optimal pH and temperature were 8.5 and 40 °C, respectively. Appropriate Zn 2+ , Mn 2+ , Mg 2+ , Tween 80, SDS, Triton X‐100, and BSA concentrations could greatly increase the activity of this enzyme. By contrast, EDTA, 1,10‐phenanthroline, NaF, and PMSF strongly inhibited its activity. The purified enzyme showed K m and V max values were 5.532 nmol/mL and 33.445 nmol/min. The CY residue in lettuce and cherry tomatoes could be removed more than 50% under the conditions of the treatment concentration for 500 mg/L and the enzyme preparation dilution of 100 times. These results suggested that the CY‐degrading enzyme, a constitutive enzyme that mainly exists in the extracellular space, was a novel esterase that might be used to detoxify CY, and could remove CY in vegetables effectively. Practical Application Our research found a novel cypermethrin‐hydrolyzing esterase from Bacillus licheniformis B‐1 and proved that the enzyme could remove cypermethrin in vegetables effectively.