NOVEL DHAP-DEPENDENT ALDOLASES FOR ORGANIC SYNTHESIS

依赖于DHAP的醛缩酶催化DHAP和醛的C-C键缩合。该反应产物作为药物或医药中间体具有很高的治疗价值。虽然这些醛缩酶具有工业应用的潜力，但到目前为止所描述的酶存在稳定性问题，并受到底物特性的限制。微生物酶在有机合成反应中的应用具有底物专一性高、反应条件温和、底物转化完全、产生化学废物少等优点。耐热酶通常提供所有这些特征以及增强的稳定性。Thermogen建议开发一套稳定的依赖于DHAP的醛缩酶和一种用于有机合成反应的高效、可扩展的技术。在第一阶段，我们将通过鉴定一组具有不同立体选择性的四种酶来证明其可行性，这些酶由具有更广泛底物特异性和稳定性的新的醛缩酶或诱变的醛缩酶组成。我们还将确定关键的反应参数来测试合成效用，并展示它们催化模型合成反应的能力。在第二阶段，我们将开发优化合成条件、酶生产和扩大底物范围的方法。在第三阶段，我们将把酶和技术商业化，供合成有机化学家和制药商使用。建议的商业应用：Thermogen建议开发新型依赖DHAP的醛缩酶，用作立体选择性有机合成反应的合成工具。醛缩酶将有助于高效、经济地合成药物或中间体，并减少化学废物的产生。

DHAP-dependent aldolases catalyze the C-C bond condensation of DHAP and an aldehyde. The products of this reaction show high therapeutic value as drugs or pharmaceutical intermediates. While these aldolases have potential for industrial applications, the enzymes described to date have stability problems and are limited by their substrate specificities. The application of microbial enzymes for organic synthetic reactions can offer high substrate specificity, complete substrate conversion under mild reaction conditions, and less chemical waste generation. Thermostable enzymes in general offer all these features as well as enhanced stability. ThermoGen proposes to develop a set of stable DHAP-dependent aldolases and an efficient, scaleable technology for use in organic synthesis reactions. In Phase I we will show feasibility by identifying a set of four enzymes with different stereoselectivities made up of either new aldolases or mutagenized aldolases with broader substrate specificities and stability. We will also identify key reaction parameters to test synthetic utility and demonstrate their ability to catalyze model synthetic reactions. In Phase II we will develop methods for optimal synthesis conditions, enzyme production, and expand the substrate range. In Phase III we will commercialize the enzymes and technology for use by synthetic organic chemists and pharmaceutical manufacturers. PROPOSED COMMERCIAL APPLICATIONS: ThermoGen proposes to develop novel DHAP-dependent aldolases for use as synthetic tools in stereoselective organic synthetic reactions. Aldolases will help syntheses or pharmaceutical drugs or intermediates efficiently and economically and reduce the chemical waste generation.