Cloning and expression of a novel esterase gene cpoA from Burkholderia cepacia

Abstract

Aims: To screen and clone a novel enzyme with specific activity for the resolution of (R)-b-acetylmercaptoisobutyrate (RAM) from (R,S)-b-acetylmercaptoisobutyrate [(R,S)-ester]. Methods and Results: A micro-organism that produces a novel esterase was isolated and identified as the bacterium Burkholderia cepacia by using the analysis of cellular fatty acids, Biolog automated microbial identification/characterization system, and 16S rRNA gene sequence analysis. A novel esterase gene was cloned from the chromosomal DNA of B. cepacia and was designated as cpoA. The cpoA encodes a polypeptide of 273 amino acids which shows a strong sequence homology with many bacterial nonhaeme chloroperoxidases. In addition, a typical serine-hydrolase motif, Gly-X-Ser-X-Gly, and the highly conserved catalytic triad, Ser95, Asp224, and His253, were identified in the deduced amino acid sequence of cpoA by multiple sequence alignment. Conclusion: The cpoA cloned from B. cepacia encodes a novel esterase which is highly related to the nonhaeme chloroperoxidases. Significance and Impact of the Study: This is the first report that describes the isolation and cloning of a serine esterase gene from B. cepacia, which is useful in the chiral resolution of (R,S)-ester. The cloned gene will allow additional research on the bifunctionality of the enzyme with esterase and chloroperoxidase activity at the structural and functional levels.

Functional Analysis of Esterase TCE2 Gene from Tetranychus cinnabarinus (Boisduval) involved in Acaricide Resistance

“…The synergistic action of several CarEs in conferring resistance against acaricides is known in some mites and ticks. For instance, it has been certified that CarEs contribute to resistance to bifenthrin, fenpyroximate, and spirodiclofen in Tetranychus urticae 11 12 13 and phoxim in Panonychus citri 14 . Significantly higher activity of esterases has been observed in resistant populations of Rhipicephalus bursa 15 .…”

MOLECULAR CHARACTERIZATION OF TWO CARBOXYLESTERASE GENES OF THE CITRUS RED MITE, Panonychus citri (ACARI: TETRANYCHIDAE)

“…The predicted amino acid sequences of PCE1 and PCE2 had highly conserved motifs of the serine hydrolase super family (Figs. and ), including serine active site (G × S × G), and the site of forming the disulfide bond (Kim et al, ). The serine active sites are different in PCE1 and PCE2 (GESAG in PCE1 and GQSAG in PCE2 ).…”

“…The synergistic action of several CarEs in conferring resistance to acaricides is known in some mites or ticks. For instance, it has been certified that CarEs contribute to resistance to bifenthrin, fenpyroximate, and spirodiclofen in Tetranychus urticae (Rauch and Nauen, ; Kim et al, ; Van Leeuwen and Tirry, ) and phoxim in P. citri (Chen et al, ). Significantly higher activity of esterases (EST) has been observed in resistant populations of Rhipicephalus bursa (Enayati et al, ).…”

Molecular cloning of levan fructotransferase gene from Arthrobacter ureafaciens K2032 and its expression in Escherichia coli for the production of difructose dianhydride IV

“…1990). Batch culture was carried out at pH 7·0 and 37°C using a rich medium (Kim et al. 2004), containing 20 g l −1 of glucose as a carbon source, and 100 μ g ml −1 of ampicillin, in a 5‐l jar fermentor (Kobiotech Co., Incheon, Korea).…”