Altschul S. F.

Chich J. F.

Desomer J.

Grzeszik C.

Heikoff S.

Holt P. J.

Johnson E. L.

Kesseler M.

Komeda H.

Meyer-Barton E. C.

Mira A. L.

Rathbone D. A.

Rorsch A.

Vesanto E.

Weisburg W. G.

Yoshpe-Besancon I.

Yüksel G. U.

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.
          </jats:p

Matthew M. Bresler

Susan J. Rosser

Amrik Basran

Neil C. Bruce

Crossref

Applied and Environmental Microbiology

Gene Cloning and Nucleotide Sequencing and Properties of a Cocaine Esterase from
            <i>Rhodococcus</i>
            sp. Strain MB1

Gene Cloning and Nucleotide Sequencing and Properties of a Cocaine Esterase from Rhodococcus

Gene Cloning and Nucleotide Sequencing and Properties of a Cocaine Esterase from Rhodococcus

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