Bresler_2000_Appl.Environ.Microbiol_66_904

Reference

Title : Gene cloning and nucleotide sequencing and properties of a cocaine esterase from Rhodococcus sp. strain MB1 - Bresler_2000_Appl.Environ.Microbiol_66_904
Author(s) : Bresler MM , Rosser SJ , Basran A , Bruce NC
Ref : Applied Environmental Microbiology , 66 :904 , 2000
Abstract : A strain of Rhodococcus designated MB1, which was capable of utilizing cocaine as a sole source of carbon and nitrogen for growth, was isolated from rhizosphere soil of the tropane alkaloid-producing plant Erythroxylum coca. A cocaine esterase was found to initiate degradation of cocaine, which was hydrolyzed to ecgonine methyl ester and benzoate; both of these esterolytic products were further metabolized by Rhodococcus sp. strain MB1. The structural gene encoding a cocaine esterase, designated cocE, was cloned from Rhodococcus sp. strain MB1 genomic libraries by screening recombinant strains of Rhodococcus erythropolis CW25 for growth on cocaine. The nucleotide sequence of cocE corresponded to an open reading frame of 1,724 bp that codes for a protein of 574 amino acids. The amino acid sequence of cocaine esterase has a region of similarity with the active serine consensus of X-prolyl dipeptidyl aminopeptidases, suggesting that the cocaine esterase is a serine esterase. The cocE coding sequence was subcloned into the pCFX1 expression plasmid and expressed in Escherichia coli. The recombinant cocaine esterase was purified to apparent homogeneity and was found to be monomeric, with an M(r) of approximately 65,000. The apparent K(m) of the enzyme (mean +/- standard deviation) for cocaine was measured as 1.33 +/- 0.085 mM. These findings are of potential use in the development of a linked assay for the detection of illicit cocaine.
ESTHER : Bresler_2000_Appl.Environ.Microbiol_66_904
PubMedSearch : Bresler_2000_Appl.Environ.Microbiol_66_904
PubMedID: 10698749
Gene_locus related to this paper: rhosm-cocE

Related information

Gene_locus related to this paper: rhosm-cocE

Citations formats

Bresler MM, Rosser SJ, Basran A, Bruce NC (2000)
Gene cloning and nucleotide sequencing and properties of a cocaine esterase from Rhodococcus sp. strain MB1
Applied Environmental Microbiology 66 :904

Bresler MM, Rosser SJ, Basran A, Bruce NC (2000)
Applied Environmental Microbiology 66 :904