Experiments were conducted to assess the ability of Streptomyces (strain PS1/5) to metabolize twelve herbicides representing several different classes including: acetanilides, triazines, ureas, uracils, and imidazoles. Incubations in aqueous culture with dextrin as carbon source and either ammonium or Casamino acids as nitrogen source resulted in transformations (> 50%) of eight of the herbicides tested: alachlor, metolachlor, atrazine, prometryne, ametryne, linuron, tebuthiuron, and bromacil; the remaining four herbicides (cyanazine, diuron, metribuzin, and imazapyr) were also transformed, but to a lesser extent. In most instances, biotransformations occurred concurrently with growth and results were consistent regardless of the nitrogen source (ammonium vs. Casamino acids). However, in some instances there were differences in rates of biotransformation as a consequence of the nitrogen source (e.g. alachlor, metribuzin), suggesting the selective induction of certain metabolic enzymes; in other instances biotransformations were not associated with growth, suggesting secondary metabolism. An experiment was also conducted to assess the ability of Streptomyces (strain PS1/5) to metabolize atrazine contaminated soil. Inoculation of soil amended with 20 micrograms/g of atrazine and 5% chitin as carbon source resulted in ca. 78% removal of atrazine within 28 days. These data suggest that Streptomyces species may be potential candidates for soil inoculation to bioremediate herbicide contaminated soils.
Currently, there has been limited use of genetic engineering for waste treatment. In this work, we are developing a procedure for the in situ treatment of toxic organophosphate wastes using the enzyme parathion hydrolase. Since this strategy is based on the use of an enzyme and not viable microorganisms, recombinant DNA technology could be used without the problems associated with releasing genetically altered microorganisms into the environment. The gene coding for parathion hydrolase was cloned into a Streptomyces lividans, and this transformed bacterium was observed to express and excrete this enzyme. Subsequently, fermentation conditions were developed to enhance enzyme production, and this fermentation was scaled-up to the pilot scale. The cell-free culture fluid (i.e., a nonpurified enzyme solution) was observed to be capable of effectively hydrolyzing organophosphate compounds under laboratory and simulated in situ conditions.
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