In vivo Selection of Ribosomal Aptamers for Improved Bioremediation

DESCRIPTION (provided by applicant): Bioremediation technologies use microorganisms to reduce, eliminate, contain, or transform to benign products contaminants present in soils, sediments, water, and air. While biological treatment of domestic and industri al waste has been employed for some time, only recently have genetic engineering approaches been applied to augment microbial capabilities. Most genetic engineering research, however, has focused on manipulation of metabolic pathways to enhance degradation rates of environmental contaminants. This project focuses on greatly enhancing the ability of microorganisms to sequester trace contaminants from wastewater treatment streams (among other possible applications) by introducing and evolving functional RNA l igands or aptamers within ribosomal RNA. Over 69 percent of the U.S. population is served by municipal wastewater treatment plants that provide secondary (i.e. biological) treatment. While such processes are generally highly effective in removing the b ulk of organic contaminants from the wastewater stream, many contaminants are refractory to degradation (chemical transformation by normal biological processes). In the extreme, metal ions are transformed by relatively few processes and physico-chemical me thods are therefore often required to polish wastewater of these contaminants. Many pesticides, pharmaceuticals, and hormones also accrue in the environment which have significant bioactivity even in trace amounts, and these contaminants are often not full y removed by conventional wastewater treatment. We will demonstrate the feasibility of our novel bacterial strains for improved removal of both organic and inorganic contaminants, including metal ions, large heterocyclic organic compounds representative of pesticides and other industrial byproducts, and hormones. The anticipated result of this study is the development of molecular biology methods and organism strains enabling a vast reduction in the release of virtually any recalcitrant molecular compound d uring biological wastewater treatment with obvious positive consequences to human health and the environment. Additionally, the developed strains are expected to have considerable commercial value for bioremediation at industrial, government, and military clean-up sites.