Development of carboxyl methyltransferases for sustainable synthesis

开发用于可持续合成的羧甲基转移酶

• 批准号: BB/W016052/1
• 负责人: Andrew Carnell
• 金额: $ 50.72万
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FW016052%2F1
• 关键词: Development; carboxyl; methyltransferases; sustainable; synthesis

The research proposed aims to provide new sustainable processes for the production of chemicals using nature's catalysts, enzymes. In particular we will focus on an enzyme that is able to catalyse the production of chemicals called esters, which are ubiquitous in chemical products such as pharmaceuticals and also important as chemical intermediates for synthetic processes. The formation of an ester from an acid often involves energy demanding processes or the use of toxic chemicals and the reactions must be done in the absence of water. The enzyme under development in this project will allow the formation of esters under aqueous conditions, making it compatible with other enzyme catalysts and allowing the development of biological synthetic pathways for chemical synthesis, or cascades. In this project, we will develop highly efficient mutants of the ester forming enzyme that are able to catalyse the formation of key intermediates for the syntheis of bioplastics and pharmaceuticals. We will use state-of-the-art screening technology that is already in use in our laboratories to identify the mutants. We will then couple the new enzymes with other smaller enzyme cascades that we have already developed for the synthesis of biodegradable bioplastics from biorenewable materials such as cellulose and lignin. We will also develop new cascades for the synthesis of pharmaceuticals by coupling with other new enzymes we have discovered recently that are surprisingly able to catalyse the formation of amide bonds in water. Amide bonds are important bonds within pharmaceuticals. The combination of the ester formation followed by the amide formation will enable replacement of expensive coupling reagents used in conventional amide bond synthesis. The ester forming enzyme needs a specific molecule found in cells called SAM to act as a catalyst. We will develop optimised cell hosts (so-called microbial cell factories) to produce all the required enzymes and the SAM, such that cell fermentations can be used to carry out the synthesis of the target chemicals. We will demonstrate a fermentation for the conversion of waste plastic, PET, into a pharmaceutical drug. We also will develop cells able to metabolise methanol that will drive the synthesis and a novel strain that can recycle the methanol produced in the amide bond forming reaction which is used by the ester forming enzyme making the whole process able to function with very small amounts of methanol.

这项研究旨在为使用自然界的催化剂酶生产化学品提供新的可持续工艺。特别是，我们将专注于能够催化称为酯的化学品生产的酶，酯在化学产品中无处不在，如药品，也是合成过程中重要的化学中间体。从酸形成酯通常涉及能量需求过程或使用有毒化学品，并且反应必须在没有水的情况下进行。该项目正在开发的酶将允许在水性条件下形成酯，使其与其他酶催化剂相容，并允许开发化学合成的生物合成途径或级联。在这个项目中，我们将开发高效的酯形成酶突变体，这些突变体能够催化生物塑料和药物合成的关键中间体的形成。我们将使用我们实验室已经使用的最先进的筛选技术来识别突变体。然后，我们将把新的酶与其他较小的酶级联反应结合起来，这些酶级联反应是我们已经开发的，用于从生物可再生材料（如纤维素和木质素）合成可生物降解的生物塑料。我们还将通过与我们最近发现的其他新酶偶联来开发用于药物合成的新级联，这些酶能够令人惊讶地催化水中酰胺键的形成。酰胺键是药物中的重要键。酯形成和随后酰胺形成的组合将使得能够替换常规酰胺键合成中使用的昂贵的偶联剂。酯形成酶需要在细胞中发现的一种称为SAM的特定分子作为催化剂。我们将开发优化的细胞宿主（所谓的微生物细胞工厂），以生产所有所需的酶和SAM，从而可以使用细胞发酵来合成目标化学品。我们将展示将废塑料PET转化为药物的发酵过程。我们还将开发能够代谢甲醇的细胞，这将驱动合成和一种新的菌株，该菌株可以回收酰胺键形成反应中产生的甲醇，该反应由酯形成酶使用，使整个过程能够用非常少量的甲醇发挥作用。