Data Mining Assisted Discovery of Novel Organophosphorou

描述（由申请人提供）：有机磷（OP）神经毒素，如对氧磷、沙林、梭曼、VX和Russian-VX，是一大类常用作化学战剂的化学品。目前，全世界储存的化学战剂超过10万吨。此外，数千吨OP化合物被用作杀虫剂，每年释放到环境中。被称为磷酸三酯酶的酶，通常被称为有机磷酸水解酶（OPH），能够将OP农药和CW试剂水解成无害的产品。有机磷酸水解酶最近已成为集中的研究工作，旨在开发这些酶的产品，酶为基础的去污。然而，目前可用的酶缺乏许多所需的性质，如溶液溶解度，热和pH稳定性和底物特异性的OPH的所有潜在的应用。许多OPH同源物可以从已经部分或完全测序的大量细菌和古细菌基因组中鉴定出来。不幸的是，即使不是大多数，也有许多是没有活性的有机磷酸酯水解酶。我们已经开发了一种数据挖掘方法，用于识别活跃的OPH。该方法已被用来发现两个新的OPH的嗜热嗜热脂肪土芽孢杆菌和硫磺硫化叶菌。目前区分活性和非活性OPH的方法取决于活性OPH的有限池大小。我们建议分离和分析一系列的五个OPH同系物，以完善我们的鉴定方法，以最大限度地提高活性OPH的池大小。此外，嗜热脂肪土芽孢杆菌和硫磺硫化叶菌酶将进一步表征（包括结晶）其在基于酶的去污系统中使用的适用性。

DESCRIPTION (provided by applicant): Organophosphorous (OP) neurotoxins such as paraoxon, sarin, soman, VX and Russian-VX, are a broad class of chemicals commonly used as chemical warfare (CW) agents. Currently, over 100,000 tons of chemical warfare agents are stockpiled worldwide. In addition, thousands-of-tons of OP compounds are used as pesticides and are annually released into the environment. Enzymes known as phosphotriesterases, commonly called organo-phosphate hydrolases (OPH's), are capable of hydrolyzing OP pesticides and CW agents into harmless products. Organophosphate hydrolases have recently become the focus of intense research efforts aimed at developing these enzymes into products for enzyme-based decontamination. However, currently available enzymes lack many of the desired properties such as solution solubility, thermal and pH stability and substrate specificity for all of the potential applications of OPH's. Numerous OPH homologues can be identified from the copious number of bacterial and archael genomes that have been either partially or completely sequenced. Unfortunately, many if not most are not active organophosphate hydrolases. We have developed a data mining method for identifying active OPH's. The method has already been used to discover two novel OPH's from the thermophiles Geobacillus stearothermophilus and Sulfolobus solfataricus. The current method of discriminating between active and inactive OPH's is dependent upon a limited pool size of active OPH's. We propose to isolate and analyze a series of five OPH homologues in order to refine our method of identification in order to maximize the pool size of active OPH's. In addition, the Geobacillus stearothermophilus and Sulfolobus solfataricus enzymes will be further characterized (including crystallization) for their suitability for use in enzyme-based decontamination systems.