The biosynthesis of N-N bond-containing natural products

含N-N键天然产物的生物合成

• 批准号: 10580666
• 负责人: Emily Patricia Balskus
• 金额: $ 34.85万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10580666
• 关键词: Anabolism; Architecture; Attention; Biochemical; Biochemistry; Biological; Biology; Breathing; Chemical Structure; Chemicals; Chemistry; Chemotherapy-Oncologic Procedure; Clinical; Clinical Trials; Complex; Drug usage; Enzymes; Future; Gene Cluster; Genes; Genome; Genomics; Goals; Health; Human; Knowledge; Life; Logic; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Medicine; Molecular; Natural Products; Nature; Nitrogen; Outcome; Pathway interactions; Pharmaceutical Preparations; Pharmacologic Substance; Proteins; Reagent; Research; Source; Streptozocin; Synthesis Chemistry; Therapeutic; Work; bioactive natural products; catalyst; chemical reaction; clinical candidate; diazo compound; functional group; guided inquiry; improved; interest; microbial; microorganism; novel; pharmacophore; programs; scaffold; small molecule; synthetic biology

PROJECT SUMMARY Microbial natural products possess complex chemical structures as well as potent biological activity and are an important source of drugs. While these molecules have captivated synthetic and medicinal chemists for decades, more recently the underlying biosynthetic pathways that construct natural product scaffolds have been recognized as important reservoirs of novel enzymes. Uncovering new enzymatic chemistry and biosynthetic strategies expands our basic understanding of Nature’s synthetic capabilities. It is also a critical first step toward applications of this fundamental knowledge and can serve as an inspiration for synthetic chemists. The long- term goal of the proposed research is to identify microbial enzymes that catalyze previously unappreciated chemical transformations. We envision discovering such enzymes by studying the biosynthesis of natural products containing important molecular architecture and functional groups of unknown biosynthetic origin. An important class of such structural motifs are functional groups containing a nitrogen-nitrogen (N–N) bond, a chemical linkage found in 9% of the 200 best-selling drugs. Reactive N–N bond-containing functional groups, including diazo and N-nitroso groups, are a critical part of biologically active small molecules including streptozotocin (Zanosar®), a clinically used treatment for metastatic pancreatic cancer. They are also uniquely enabling chemical reagents, with diazo compounds mediating many important and powerful chemical transformations in synthetic chemistry, biocatalysis, and biorthogonal chemistry. Though reactive N–N bonds are present in microbial natural products, their biosynthetic origins are poorly understood. Thus, the overall objective of this application is to discover and characterize enzymes that construct diazo- and N-nitroso- containing metabolites. Preliminary results from our lab and others have uncovered biosynthetic gene clusters responsible for constructing multiple diazo- and N-nitroso-containing natural products, including streptozotocin and other molecules that have been in clinical trials. These findings set the stage for our three complementary specific aims: 1) identify and characterize the biosynthetic enzymes responsible for constructing the diazo groups of the natural products cremeomycin and kinamycin; 2) identify and characterize the biosynthetic enzymes responsible for constructing the N-nitroso groups of the natural products streptozotocin and alanosine; 3) access additional diazo and N-nitroso biosynthetic enzymes and natural products by characterizing cryptic gene clusters. By leveraging the tremendous structural diversity of microbial natural products in the genomic era, we will rapidly discover and characterize biosynthetic transformations that fill critical gaps in our current knowledge of enzymatic chemistry capabilities. Finally, the workflow we have formulated for investigating the biosynthesis of reactive N– N bond-containing functional groups will also be readily generalizable to additional structural motifs found in microbial natural products.

项目摘要 微生物天然产物具有复杂的化学结构以及潜在的生物学活性，是一种 药物的重要来源。尽管这些分子吸引了数十年来吸引合成和医学化学家 最近，构建天然产物支架的基本生物合成途径已经 被认为是新型酶的重要储层。发现新的酶学化学和生物合成 策略扩大了我们对自然合成能力的基本理解。这也是迈向的关键第一步 这种基本知识的应用，可以作为合成化学家的灵感。长期 拟议研究的术语目标是鉴定微生物酶，这些酶催化先前未批准的酶 化学转化。我们通过研究自然的生物合成来设想发现这种酶 包含重要分子结构和未知生物合成起源的官能团的产品。一个 重要类别的结构基序是含有氮氮（N – N）键的官能团 在200种最畅销药物中，有9％的化学连接发现。反应性n – N含键的官能团， 包括重氮和N-亚硝基组，是生物活性小分子的关键部分 链霉菌素（Zanosar®），一种用于转移性胰腺癌的临床治疗。他们也很独特 启用化学试剂，并带有重氮化合物，介导了许多重要且功能强大的化学物质 合成化学，生物催化和生物三相化学的转化。虽然反应性N – N键 它们存在于微生物天然产物中，其生物合成起源知之甚少。那，总体 该应用的目的是发现和表征构建重氮和N-硝基的酶 含有代谢物。我们实验室和其他人的初步结果已经发现了生物合成基因簇 负责构建含有含链霉菌素的多重量和N-硝基的天然产物 以及其他正在临床试验的分子。这些发现为我们的三个完成奠定了舞台 具体目的：1）识别和表征负责构建重氮组的生物合成酶 天然产物丝霉素和kinamycin; 2）识别和表征生物合成酶 负责构建天然产物链霉菌素和丙氨酸的N-亚硝酸基团； 3）访问 通过表征隐源性簇来表征其他重氮和N-亚硝基生物合成酶和天然产物。 通过利用基因组时代微生物天然产物的巨大结构多样性，我们将迅速 发现并表征生物合成转化，这些转化填补了我们当前对酶促的知识的关键空白 化学能力。最后，我们为研究反应性n –的生物合成而制定的工作流程 n含键的官能团也将很容易被推广到在 微生物天然产品。