Synthetic biology approaches to construct metal analogues of vitamin B12 to act as anti-microbial and imaging agents for health applications.

利用合成生物学方法构建维生素 B12 的金属类似物，作为健康应用的抗菌剂和显像剂。

• 批准号: 2881504
• 金额: --
• 依托单位: University of Southampton
• 依托单位国家: 英国
• 项目类别: Studentship
• 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=studentship-2881504
• 关键词: Synthetic; biology; approaches; construct; metal

Vitamin B12 (cobalamin) is an essential micronutrient whose synthesis is restricted to only certain bacteria. It is one of the most complex molecules found in Nature, consisting of a cobalt-containing corrin ring and a lower base attached via the nucleotide loop. The biological activity of the molecule is dependent on the properties of the metal ion at its centre.We have successfully prepared several metal analogues of cobalamin (Rh, Zn and Ni). This was achieved using a novel approach consisting of an engineered bacterial strain which can produce a metal-free intermediate, hydrogenobyric acid (Hby), and then subsequent biochemical and chemical steps to complete the synthesis and produce the desired analogues. While the system works well, we are looking to optomise/simplify the synthesis by further engineering the E. coli strain, using genes from Chromatium vinosum which is known to produce cobalt-free cobalamin under specific conditions.We have determined that the rhodium analogue is an inhibitor of B12-dependent processes, but we currently know very little about the biological activity of the metal analogues. We wish to expand our knowledge in this area by looking at the cellular uptake and trafficking of the analogues, their effect on B12-dependent enzymes and processes, and document any antimicrobial/anticancer activity present. The ability to replace the metal ion also opens the possibility of creating unique medical imaging agents for positron emission tomography (PET) which could be used in cancer diagnosis.The aim of this project is to:1. Synthesise a range of metal analogues of cobalamin (Cu, Rh, Zn and Ni).2. Investigate the biological properties of the metal analogues (eg. vitamin uptake/transport, enzyme inhibition, antimicrobial/anticancer agents).3. Engineer an E. coli strain capable of producing cobalt-free cobalamin (hydrogenobalamin).

维生素B12(钴胺)是一种必需的微量营养素，其合成仅限于某些细菌。它是自然界中发现的最复杂的分子之一，由一个含钴的科林环和一个通过核苷酸环连接的下部碱基组成。该分子的生物活性取决于其中心金属离子的性质。我们已经成功地制备了钴胺的几种金属类似物(Rh，Zn和Ni)。这是通过一种新的方法实现的，该方法包括一种工程菌，它可以产生一种不含金属的中间体--氢比酸(Hby)，然后通过后续的生化和化学步骤来完成合成并生产所需的类似物。虽然该系统运行良好，但我们希望通过进一步改造大肠杆菌菌株来优化/简化合成，使用已知的在特定条件下产生无钴钴胺的棕色染色杆菌的基因。我们已经确定Rh类似物是B12依赖过程的抑制物，但目前我们对金属类似物的生物活性知之甚少。我们希望通过观察细胞对类似物的摄取和运输，它们对B12依赖的酶和过程的影响，并记录存在的任何抗菌/抗癌活性，来扩大我们在这一领域的知识。取代金属离子的能力也开启了为正电子发射断层扫描(PET)创造独特的医学显像剂的可能性，可以用于癌症诊断。本项目的目标是：1.合成一系列钴的金属类似物(铜、Rh、锌和镍)。研究金属类似物的生物学特性(例如。维生素摄取/转运、酶抑制、抗菌/抗癌药物)。设计一种能够生产无钴钴胺(氢钴胺)的大肠杆菌菌株。