In vivo Selection of Ribosomal Aptamers for Improved Bioremediation

描述(由申请人提供)：生物修复技术使用微生物来减少、消除、包含或转化为土壤、沉积物、水和空气中存在的无害产品污染物。虽然生物处理生活和工业废物已经有一段时间了，但直到最近才应用基因工程方法来增强微生物的能力。然而，大多数基因工程研究都集中在操纵代谢途径以提高环境污染物的降解率。该项目的重点是通过在核糖体RNA中引入和进化功能性RNA配体或适配子，极大地提高微生物从废水处理流中隔离微量污染物的能力(以及其他可能的应用)。超过69%的美国人口由市政污水处理厂提供服务，这些污水处理厂提供“二级”(即生物)处理。虽然这类工艺在去除废水中的大部分有机污染物方面通常非常有效，但许多污染物难以降解(通过正常的生物工艺进行化学转化)。在极端情况下，金属离子的转化过程相对较少，因此通常需要物理化学方法来抛光这些污染物的废水。许多农药、药品和激素也会在环境中积累，即使是微量的，也具有显著的生物活性，而这些污染物通常不能通过传统的废水处理方法完全去除。我们将展示我们的新型细菌菌株改善有机和无机污染物去除的可行性，包括金属离子、以杀虫剂和其他工业副产品为代表的大杂环有机化合物以及激素。这项研究的预期结果是发展分子生物学方法和生物体菌株，使生物废水处理过程中几乎任何顽固分子化合物的释放大大减少，对人类健康和环境产生明显的积极影响。此外，开发的菌株有望在工业、政府和军事清理场所的生物修复方面具有相当大的商业价值。

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 industrial 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 ligands 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 bulk 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 methods 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 fully 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 during 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.