DEGRADATION OF COCAINE BY A MIXED CULTURE OF PSEUDOMONAS-FLUORESCENS MBER AND COMAMONAS-ACIDOVORANS MBLF

A mixed culture that could utilize cocaine as the sole source of carbo n and energy for growth was isolated by selective enrichment, The indi vidual microorganisms within this mixed culture were identified as Pse udomonas fluorescens (termed MBER) and Comamonas acidovorans (termed M BLF). Each microorganism was shown to be unable to grow to any appreci able extent on 10 mM cocaine in the absence of the other, C. acidovora ns MBLF was found to possess an inducible cocaine esterase which catal yzed the hydrolysis of cocaine to ecgonine methyl ester and benzoate, C. acidovorans was capable of growth on benzoate at concentrations bel ow 5 mM but was unable to metabolize ecgonine methyl ester, P. fluores cens MBER was capable of growth on either benzoate as the sole source of carbon or ecgonine methyl ester as the sole source of carbon and ni trogen, P, fluorescens MBER was found to initiate the degradation of e cgonine methyl ester via ecgonine, pseudoecgonine, and pseudoecgonyl-c oenzyme A. Subcellular studies resulted in the identification of an ec gonine methyl esterase, an ecgonine epimerase, and a pseudoecgonyl-coe nzyme A synthetase which were induced bg growth on ecgonine methyl est er or ecgonine. Further metabolism of the ecgonine moiety is postulate d to involve nitrogen debridging, with the production of carbonyl-cont aining intermediates.