Understanding the specificity and non-native catalytic activity of PLP-dependent enzymes

了解 PLP 依赖性酶的特异性和非天然催化活性

• 批准号: 10715757
• 负责人: Yang Hai
• 金额: $ 37.74万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA BARBARA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715757
• 关键词: Amines; Amino Acids; Biology; Carbon; Cell physiology; Chemicals; Complex; Data; Enzymatic Biochemistry; Enzymes; Evolution; Family; Goals; Health; Human; Investigation; Knowledge; Laboratories; Metabolism; Nature; Pharmacologic Substance; Physiology; Play; Research; Role; Specificity; Structure; Substrate Specificity; Therapeutic; Work; aminoacid biosynthesis; bioactive natural products; frontier; non-Native; novel; programs; tool

Project Summary PLP-dependent enzymes are one of the most versatile biocatalysts and catalyze a diverse range of chemical transformations. They are widespread in nature and play critical roles in metabolism and numerous cellular processes. Studying PLP enzymes is hence important for us to understand biology and develop therapeutics. Because of their exquisite and versatile catalytic activity, PLP-dependent enzymes are also remarkable biocatalysts to build diverse structurally complex and bioactive natural products; and are indispensable biocatalytic tools for asymmetric synthesis of noncanonical amino acids and chiral amine pharmaceuticals. However, despite the vast number of PLP-dependent enzymes characterized to date, our abilities to predict, manipulate, and harness their activities are still largely limited. This research program desires to fill the knowledge gap by integrating discovery, mechanistic investigation, and biocatalytic application to systematically and comprehensively study four types of carbon-carbon (C-C) bond forming and cleaving PLP enzymes, including our recently discovered PLP-dependent Mannich cyclase. These enzymes represent the frontier of PLP enzymology because of their unusual activity, complementary synthetic utility to existing biocatalysts, and unexpected evolutionary relationship with well-characterized PLP enzyme family. All proposed aims are supported by strong preliminary data gathered in our laboratory. Our overarching goal is to understand the chemical and substrate specificity and leverage this understanding to uncover previously unknown functions of PLP-dependent enzymes and explore their non-native catalytic utility. Ultimately, the proposed research will expand our mechanistic understanding on PLP enzymology, shed new light on metabolism, and provide novel biocatalytic tools for amino acid biosynthesis.

项目摘要 PLP依赖性酶是最通用的生物催化剂之一，并催化了多样的范围 化学转化。它们在自然界广泛，在代谢中起着关键作用和许多 细胞过程。因此，研究PLP酶对我们了解生物学和发展很重要 疗法。由于其精致且通用的催化活性，PLP依赖性酶也是 出色的生物催化剂，以建立各种结构复杂和生物活性天然产品；是 不可或缺的生物催化工具，用于非对称氨基酸和手性胺的不对称合成 药品。然而，尽管迄今为止有大量的PLP依赖性酶，但我们的 预测，操纵和利用其活动的能力仍然很大程度上受到限制。该研究计划希望 通过将发现，机械研究和生物催化应用集成到知识差距 系统，全面研究四种类型的碳碳（C-C）键形成和切割PLP 酶，包括我们最近发现的PLP依赖性Mannich Cyclase。这些酶代表 PLP酶学的前沿由于其非同寻常的活性，因此与现有的互补合成效用 生物催化剂以及与特征良好的PLP酶家族的意外进化关系。都提出了 目的是由我们实验室收集的强大初步数据支持的。我们的总体目标是了解 化学和底物特异性并利用这种理解来揭示以前未知功能 依赖PLP的酶并探索其非本地催化效用。最终，拟议的研究将 扩展我们对PLP酶学的机械理解，为新陈代谢提供新的启示，并提供新颖 氨基酸生物合成的生物催化工具。