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.

项目概要 PLP 依赖性酶是最通用的生物催化剂之一，可催化多种 化学转变。它们在自然界中广泛存在，在新陈代谢和许多 细胞过程。因此，研究 PLP 酶对于我们了解生物学和开发非常重要 疗法。由于其精致且多功能的催化活性，PLP 依赖性酶也被广泛应用。 卓越的生物催化剂，可构建多种结构复杂且具有生物活性的天然产物；并且是 非经典氨基酸和手性胺不对称合成不可或缺的生物催化工具 药品。然而，尽管迄今为止已鉴定出大量 PLP 依赖性酶，但我们的 预测、操纵和利用他们的活动的能力在很大程度上仍然有限。该研究计划希望 通过整合发现、机制研究和生物催化应用来填补知识空白 系统、全面地研究四种类型的碳-碳（C-C）键形成和断裂PLP 酶，包括我们最近发现的 PLP 依赖性曼尼希环化酶。这些酶代表 PLP 酶学的前沿，因为它们具有不寻常的活性，可补充现有的合成效用 生物催化剂，以及与已充分表征的 PLP 酶家族的意外进化关系。全部提议 我们实验室收集的强有力的初步数据支持了这些目标。我们的首要目标是了解 化学和底物特异性，并利用这种理解来揭示以前未知的功能 PLP 依赖性酶的研究并探索其非天然催化效用。最终，拟议的研究将 扩展我们对 PLP 酶学机制的理解，为新陈代谢提供新的思路，并提供新的 用于氨基酸生物合成的生物催化工具。