TRICHODIENE SYNTHASE

单孢二烯合酶

• 批准号: 7602294
• 负责人: Lakshmi Sangeetha Vedula
• 金额: $ 0.29万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7602294
• 关键词: Active Sites; Affect; Alcohols; Antibiotics; Aspartate; Binding; Biological; Complex; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Funding; Goals; Grant; Hydrocarbons; Institution; Life; Ligand Binding; Ligands; Light; Metals; Mutation; Perfume; Research; Research Personnel; Resources; Sesquiterpenes; Site; Source; Structure; United States National Institutes of Health; base; commercial application; design; enzyme activity; enzyme mechanism; farnesyl pyrophosphate; mutant; novel; trichodiene; trichodiene synthase

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Trichodiene synthase (TS) is a Mg(II)-dependent sesquiterpene cyclase. Sesquiterpene cyclases catalyze the conversion of a universal acyclic substrate farnesyl diphosphate to one of numerous cyclic and chiral sesquiterpene hydrocarbons or alcohols with exquisite geometric and stereochemical precision. These sesquiterpenes are the precursors of several molecules that are of great importance to us in our everyday life as antibiotics, culinary additives, perfumes etc. TS evolved to exclusively form only trichodiene. However, conservative site-specific mutations in the active site of the enzyme not only affect the activity of the enzyme but also derail the enzyme into making additional products that resemble the intermediates more than the product or the substrate. Therefore, the study of these products and how a certain mutation makes certain aberrant products sheds light on the mechanism of the enzyme. We are studying the structural basis for aberrant product formation by the study of the mutant TS enzymes and their complexes with various ligands. Based on the structures of various mutants, we found that the aberrant product formation is mainly a function of the active site that does not form a perfectly tailored template for the exclusive formation of trichodiene. The volumes of ligand-bound active sites are generally much larger in the mutants than in the wild-type enzyme. We have studied an aspartate-rich motif that is conserved across sesquiterpene cyclases from all species. We are currently studying the importance of a second metal-binding motif, often called the NSE (or DTE) motif. The ultimate goal of these studies on this and other sesquiterpene cyclases is to be able to predict the products of a given sesquiterpene cyclases and to design novel sesquiterpenes that will form novel sesquiterpenes with various biological and commercial applications.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Trichodiene 合酶 (TS) 是一种 Mg(II) 依赖性倍半萜环化酶。倍半萜环化酶催化通用无环底物法呢基二磷酸转化为众多环状和手性倍半萜烃或醇中的一种，具有精致的几何和立体化学精度。这些倍半萜烯是几种分子的前体，这些分子在我们的日常生活中非常重要，如抗生素、烹饪添加剂、香料等。TS 演变为仅形成单族二烯。然而，酶活性位点的保守位点特异性突变不仅影响酶的活性，而且使酶脱轨，产生与中间体比产物或底物更相似的其他产物。因此，对这些产物以及某种突变如何产生某些异常产物的研究有助于阐明酶的机制。我们正在通过研究突变型 TS 酶及其与各种配体的复合物来研究异常产物形成的结构基础。基于各种突变体的结构，我们发现异常产物的形成主要是活性位点的功能，该活性位点没有形成用于单端孢二烯排他性形成的完美定制模板。突变体中配体结合活性位点的体积通常比野生型酶大得多。我们研究了一个富含天冬氨酸的基序，该基序在所有物种的倍半萜环化酶中都是保守的。我们目前正在研究第二种金属结合基序的重要性，通常称为 NSE（或 DTE）基序。这些关于这种倍半萜环化酶和其他倍半萜环化酶的研究的最终目标是能够预测给定倍半萜环化酶的产物并设计新型倍半萜，其将形成具有各种生物和商业应用的新型倍半萜。