Evolving a stereochemically promiscuous aldolase to create new classes of stereoselective biocatalyst for asymmetric synthesis

进化立体化学混杂的醛缩酶以创建用于不对称合成的新型立体选择性生物催化剂

• 批准号: BB/E020704/1
• 负责人: Steven Bull
• 金额: $ 51.07万
• 依托单位: University of Bath
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2008
• 资助国家: 英国
• 起止时间: 2008 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE020704%2F1
• 关键词: Evolving; stereochemically; promiscuous; aldolase; create

This proposal describes the use of a thermostable enzyme to prepare new types of chiral compounds as building blocks for synthesising important intermediates for the pharmaceutical and fine chemical industry. The scope and limiation of the enzyme to catalyse a specific reaction with a wide range of substrates will be investigated. The enzyme contains an active site where catalysis takes place, and its structure has been characterised at a molecular level by X-ray crystallography. Knowledge of the structure of this active site will enable us to generate new 'mutant' catalysts with their own specificity profiles as improved biocatalysts for synthesis. These mutation studies will fall into two categories: firstly, we will deliberately introduce specific changes into the enzyme; and secondly, we will generate random mutations to afford large libraries of catalysts. Screening protocols will be developed that will enable us to rapidly identify mutants with useful catalytic activity, with the aim of identifying biocatalysts that carry out reactions not currently observed in nature. Concurrently, we will also develop a chemical approach to the synthesis of these type of compounds, using small chiral molecules as alternative catalysts. A number of compounds produced in this study will then be converted into novel highly fuctionalised intermediates of use for the synthesis of biologically-important natural products.

该提案描述了使用热稳定性酶来制备新型手性化合物，作为合成制药和精细化工行业重要中间体的基础材料。将研究酶与多种底物催化特定反应的范围和限制。该酶含有一个发生催化作用的活性位点，其结构已通过 X 射线晶体学在分子水平上进行了表征。了解该活性位点的结构将使我们能够生成具有自身特异性的新“突变”催化剂，作为改进的合成生物催化剂。这些突变研究将分为两类：首先，我们会刻意向酶中引入特定的变化；其次，我们将产生随机突变以提供大型催化剂库。我们将制定筛选方案，使我们能够快速识别具有有用催化活性的突变体，目的是识别能够进行目前在自然界中未观察到的反应的生物催化剂。同时，我们还将开发一种化学方法来合成此类化合物，使用小手性分子作为替代催化剂。本研究中产生的许多化合物将被转化为新型高度功能化的中间体，用于合成具有生物学意义的天然产物。

项目成果

Structurally informed site-directed mutagenesis of a stereochemically promiscuous aldolase to afford stereochemically complementary biocatalysts.

• DOI: 10.1021/ja104412a
• 发表时间: 2010-08
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: S. Royer;Luke J. Haslett;S. Crennell;D. Hough;M. Danson;S. Bull

The structure of Sulfolobus solfataricus 2-keto-3-deoxygluconate kinase.

硫磺菌硫化叶菌 2-酮-3-脱氧葡萄糖酸激酶的结构。

• DOI: 10.1107/s0907444908036111
• 发表时间: 2008
• 期刊: Acta crystallographica. Section D, Biological crystallography
• 作者: Potter JA