Enzymic Detoxification for Organophosphate Nerve Agents

描述（由申请人提供）：有机磷酸酯和有机膦酸酯是迄今为止合成的毒性最大的化合物之一。这些化合物是有毒的，因为它们通过快速和不可逆地灭活乙酰胆碱酯酶而干扰神经传递的固有能力。有机磷的毒性特性已被用作农业杀虫剂和化学武器。已经鉴定了一种细菌磷酸三酯酶，其可以作为催化剂用于识别、水解和解毒广谱有机磷酸盐神经毒剂。本申请的主要目的是设计和表征细菌磷酸三酯酶的修饰形式，其被优化用于选择性识别和破坏对人类健康构成最严重威胁的那些有机磷酸酯。将构建突变酶的合理和组合文库，并通过新的筛选和选择程序鉴定具有增强的催化能力的那些变体。细菌磷酸三酯酶的氨基酸序列中的渐进变化将与催化常数的增强和活性位点内的修饰直接相关，如通过X-射线衍射方法测定的。公共卫生相关性：有机磷神经毒剂的毒性对文明社会的健康和福祉构成严重威胁。该项目的主要目标是创造新的催化酶，优化其检测，破坏和解毒广谱有机磷神经毒剂的能力。

DESCRIPTION (provided by applicant): Organophosphate and organophosphonate esters are among the most toxic compounds that have ever been synthesized. These compounds are toxic because of their inherent ability to interfere with nerve transmission through the rapid and irreversible inactivation of the enzyme acetyl cholinesterase. The toxic properties of the organophosphates have been exploited as agricultural insecticides and as chemical weapons. A bacterial phosphotriesterase enzyme has been identified that can function as a catalyst for the recognition, hydrolysis, and detoxification of a broad spectrum of organophosphate nerve agents. The primary objective of this application is to design and characterize modified forms of the bacterial phosphotriesterase that are optimized for the selective recognition and destruction of those organophosphates that pose the most serious threats to human health. Rational and combinatorial libraries of mutant enzymes will be constructed and those variants with enhanced catalytic proficiency will be identified through novel screening and selection procedures. The progressive changes in the amino acid sequence of the bacterial phosphotriesterase will be directly correlated with the enhancements in the catalytic constants and the modifications within the active site as determined by X-ray diffraction methods. PUBLIC HEALTH RELEVANCE: The toxic properties of the organophosphate nerve agents represent a serious threat to the health and well being of civilized societies. The primary objective of this project is to create novel catalytic enzymes that are optimized in their ability to detect, destroy and detoxify a broad spectrum of organophosphate nerve agents.