Molecular cloning, purification and biochemical characterization of a novel pyrethroid-hydrolyzing carboxylesterase gene from Ochrobactrum anthropi YZ-1

J Hazard Mater. 2012 Jun 30:221-222:206-12. doi: 10.1016/j.jhazmat.2012.04.031. Epub 2012 Apr 20.

Abstract

Strain YZ-1 was isolated from activated sludge and identified as Ochrobactrum anthropi. This strain was capable of degrading pyrethroids pesticides, suggesting the presence of degrading enzymes. In the present study, a novel esterase gene pytZ was cloned from the genomic library of YZ-1 successfully. The pytZ contained an open reading frame of 606bp encoding a pyrethroid-hydrolyzing carboxylesterase. Deduced amino acid sequence showed moderate identities (39-59%) with most homologous carboxylesterase, except a putative carboxylesterase from O. anthropi ATCC 49188 with the highest identity of 85%. Phylogenetic analysis revealed that PytZ belonged to esterase VI family. The gene pytZ showed no any sequence similarity with reported pyrethroid-hydrolyzing genes and was a new pyrethroid-degrading gene. PytZ was expressed in Escherichia coli BL21 (DE3) and purified using Ni-NTA Fast Start. PytZ was able to degrade various pyrethroids. The optimal temperature and pH were 35°C and 7.5. This enzyme was very stable over a wide range of temperature and pH. No cofactors were required for enzyme activity. Broad substrate specificity, high enzyme activity, and the favorable stability make the PytZ a potential candidate for the detoxification of pyrethroid residues in biotechnological application.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Carboxylesterase / chemistry
  • Carboxylesterase / genetics*
  • Chromatography, Gas
  • Cloning, Molecular
  • DNA Primers
  • Escherichia coli / genetics
  • Genes, Bacterial*
  • Hydrolysis
  • Molecular Sequence Data
  • Ochrobactrum anthropi / enzymology
  • Ochrobactrum anthropi / genetics
  • Ochrobactrum anthropi / isolation & purification*
  • Polymerase Chain Reaction
  • Pyrethrins / metabolism*
  • Sequence Homology, Amino Acid
  • Substrate Specificity

Substances

  • DNA Primers
  • Pyrethrins
  • Carboxylesterase