Novel Organophosphorous Hydrolases for Decontamination

用于净化的新型有机磷水解酶

基本信息

批准号：
6834554
负责人：
ANDREW D MESECAR
金额：
$ 9.62万
依托单位：
LYBRADYN, INC.
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-20 至 2005-02-19
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6834554
关键词：
bacterial proteins bioreactors biotechnology bioterrorism /chemical warfare biotransformation directed evolution environmental toxicology enzyme activity enzyme structure esterase gram positive bacteria hazardous substances mass tissue /cell culture nerve gas neurotoxins organophosphorus insecticide technology /technique development transfection /expression vector

项目摘要

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 pesticides and chemical warfare (CW) agents. Currently, over 100,000 tons of chemical warfare agents are stockpiled worldwide, and thousands-of-tons of OP pesticides are released into the environment annually. Enzymes known as phosphotriesterases, commonly called organophosphorous hydrolases (OPH), are capable of hydrolyzing OP and CW agents into harmless products. Because of their potential for destruction of these neurotoxins, organophosphorous hydrolases have recently become the focus of intense research efforts aimed at developing these enzymes into products for enzyme-based decontamination. However, these 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 these enzymes. We have recently identified, cloned, expressed, purified, crystallized, and determined the x-ray structure of a novel OPH from an extremophile that has significant activity against the chemical warfare agent soman. The enzyme can be overexpressed in large quantities, and has desirable physical properties such as high solubility and thermostability compared to all of the other OPH enzymes under development. Therefore, our version of OPH holds significant promise for technological advancement in both civilian and military applications that require novel OPH enzymes for enzyme-based decontamination. Since large-scale expression and production using our current expression vectors and host strains would not be economically viable, and since our version of OPH has low activity toward V-agents, we propose to develop an economically viable, industrial scale production system for D. radiodurans OPH enzymes, and to enhance the catalytic potential of D. radiodurans OPH towards V-agents and other organophosphorous compounds using directed evolution genetic technology.

描述（由申请人提供）：有机磷 (OP) 神经毒素，例如对氧磷、沙林、梭曼、VX 和俄罗斯 VX，是一类广泛用作杀虫剂和化学战 (CW) 制剂的化学品。目前，全球储存的化学战剂超过10万吨，每年有数千吨有机磷农药被释放到环境中。磷酸三酯酶通常称为有机磷水解酶 (OPH)，能够将 OP 和 CW 制剂水解成无害的产品。由于有机磷水解酶具有破坏这些神经毒素的潜力，因此最近已成为大量研究工作的焦点，旨在将这些酶开发成基于酶的净化产品。然而，这些酶缺乏许多所需的特性，例如溶液溶解度、热稳定性和 pH 稳定性以及这些酶的所有潜在应用的底物特异性。我们最近从一种极端微生物中鉴定、克隆、表达、纯化、结晶并确定了一种新型 OPH 的 X 射线结构，该极端微生物对化学战剂梭曼具有显着的活性。该酶可以大量过度表达，并且与所有其他正在开发的 OPH 酶相比，具有理想的物理特性，例如高溶解度和热稳定性。因此，我们的 OPH 版本对于民用和军事应用领域的技术进步具有重大前景，这些应用需要新型 OPH 酶来进行基于酶的去污。由于使用我们当前的表达载体和宿主菌株进行大规模表达和生产在经济上不可行，并且由于我们的 OPH 版本对 V 剂活性较低，因此我们建议开发一种经济上可行的工业规模生产系统。耐辐射 D. 耐辐射 OPH 酶，并利用定向进化遗传技术增强耐辐射 D. 耐辐射 OPH 对 V 剂和其他有机磷化合物的催化潜力。