Self-sufficient reductive dehalogenases for bioremediation

用于生物修复的自给自足的还原脱卤酶

• 批准号: BB/X007952/1
• 负责人: David Leys
• 金额: $ 66.85万
• 依托单位: University of Manchester
• 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%2FX007952%2F1
• 关键词: Self; sufficient; reductive; dehalogenases; bioremediation

Halogenated organic compounds have found widespread use in the chemical and pharmaceutical industries, in agriculture, and as solvents and materials, with production on the multi-ton scale per annum. Their persistent nature and (un)controlled release has led to worldwide accumulation of such compounds in the environment. Furthermore, bioaccumulation and toxicity has led to many organohalides presenting significant challenges to human and environmental health. Indeed, the majority of persistent organic pollutants (POPs) contain halogens, with many of them now falling under the 2004 Stockholm convention that serves to eliminate their use. While regulation has reduced some levels of contamination, remaining reservoirs in sediments and waste remain of concern. Furthermore, few alternatives are available for the use of organohalides, and society is balancing human benefit with environmental detriment when regulating organohalide production and use. However, significant knowledge gaps remain in understanding the (long-term) effects of human/environmental exposure. Counter to popular perception, organohalides are not exclusively of anthropogenic origin, with an ever-increasing amount of natural compounds identified, the majority of biological origin. Indeed, the last decades have seen identification and characterisation of a range of halogenase and dehalogenase enzymes that feature in organohalide biochemistry. While dehalogenases can serve in bioremediation or biosensing of POPs, the inherent properties of these enzymes do not match the human-made xenobiotic repertoire. We seek to study a particular type of dehalogenases, the reductive dehalogenases (Rdh) or organohalide reductases, which present unique opportunities in terms of substrate scope, bioremediation and biosensing. In recent years, our own group and that of others have provided detailed insights into these enigmatic proteins that depend on vitamin B12 for activity. However, the exact mode of operation remains somewhat enigmatic, while protein engineering efforts aimed at generating robust biocatalyst based on the Rdh-template have yet to meet with success. We seek to use specific reductive dehalogenases selected and studied in our lab to provide a complete fundamental insight into the mechanism of action. We seek this information to guide directed evolution experiments aimed at generating a toolbox of dehalogenases that can act on POPs and related compounds of interest.

已发现卤化有机化合物广泛用于化学和制药工业、农业中以及作为溶剂和材料，每年的生产规模为数吨。它们的持久性和（不）受控释放导致了这些化合物在环境中的全球积累。此外，生物累积性和毒性导致许多有机卤化物对人类和环境健康构成重大挑战。事实上，大多数持久性有机污染物（POP）含有卤素，其中许多现在属于2004年斯德哥尔摩公约，该公约旨在消除其使用。虽然管制措施在一定程度上减少了污染，但沉积物和废物中剩余的水库仍然令人关切。此外，有机卤化物的使用几乎没有替代品，社会在管制有机卤化物的生产和使用时，正在平衡人类利益与环境损害。然而，在了解人类/环境接触的（长期）影响方面仍然存在重大的知识差距。与流行的看法相反，有机卤化物并不完全是人为来源的，已确定的天然化合物数量不断增加，其中大多数是生物来源的。事实上，在过去的几十年里，已经看到了一系列的卤化酶和脱卤酶的有机卤化物生物化学功能的鉴定和表征。虽然脱卤酶可用于持久性有机污染物的生物修复或生物传感，但这些酶的固有特性与人造异生素库不匹配。我们试图研究一种特殊类型的脱卤酶，还原脱卤酶（Rdh）或有机卤化物还原酶，这在底物范围，生物修复和生物传感方面提供了独特的机会。近年来，我们自己的团队和其他团队对这些依赖维生素B12进行活性的神秘蛋白质提供了详细的见解。然而，确切的操作模式仍然有些神秘，而蛋白质工程的努力，旨在产生强大的生物催化剂的基础上Rdh-template尚未取得成功。我们试图使用特定的还原脱卤酶选择和研究在我们的实验室提供一个完整的基本洞察的作用机制。我们寻求这些信息来指导定向进化实验，旨在产生一个工具箱的脱卤酶，可以对持久性有机污染物和相关化合物的利益。