Total Synthesis and Biosynthesis of Bioactive Substances

生物活性物质的全合成与生物合成

• 批准号: 8761664
• 负责人: DAVID H SHERMAN
• 金额: $ 26.85万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-15 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8761664
• 关键词: Aldehydes; Alder plant; Alkaloids; Amino Acids; Anabolism; Anti-Infective Agents; Antineoplastic Agents; Applications Grants; Aspergillus; Award; Biochemical; Biodiversity; Biogenesis; Bioinformatics; Biological; Biological Assay; Biological Factors; Biomimetics; Cells; Chemicals; Chicago; Complex; Cyclization; Dehydration; Denmark; Dimethylallyltranstransferase; Dipeptides; Enzymes; Family; Family member; Gene Cluster; Genes; Genome; Genomics; Goals; Growth; Health; Indole Alkaloids; Isoprene; Japan; Label; Laboratories; Malignant Neoplasms; Mediating; Methods; Michigan; Mining; Molecular; Molecular Genetics; National Human Genome Research Institute; Nature; Oxidases; Oxidoreductase; Parasitic Diseases; Pathway interactions; Phase; Pipecolic Acids; Preparation; Production; Proteins; Pyrans; Reaction; Recombinants; Resolution; Roentgen Rays; Series; Source; Structure; System; Technology; Tryptophan; Universities; Work; X-Ray Crystallography; base; catalyst; chemical addition; chemical synthesis; cycloaddition; design; enantiomer; fascinate; genome analysis; genome sequencing; interest; mevalonate; novel; paraherquamide; prenylation; progenitor; programs; prolylisoleucine; research study; tool; tryptophyl-proline

DESCRIPTION (provided by applicant): This revised grant application is focused on experimental studies to elucidate the biosynthesis of three biogenetically related families of natural products: (1) the Paraherquamides, Asperparalines, Malbranchemaides, Marcfortines and Chrysogenamide (the monooxopiperazines); (2) the Stephacidins, Notoamides, Waikialoids and Brevianamides (the dioxopiperazines); and (3) the Citrinadins, Citrinalins, Cyclopiamines and PF1270 alkaloids (the decarbonylated alkaloids). The goals are to exploit the powerful synergies of total synthesis, whole genome sequencing, bioinformatics analysis, genome mining, functional expression and X-ray crystallography of biosynthetic enzymes to fully elucidate the corresponding biosynthetic pathways to all three families of alkaloids. I. Paraherquamides, Asperparalines, Malbranchemaides, Marcfortines and Chrysogenamide: the Monooxopiperazines. Provocative evidence has been elucidated in our laboratory indicating that this class of alkaloids, are constructed by a rare biosynthetic intramolecular [4+2] cycloaddition reaction resulting from the reductive release of a Trp-Pro (or Trp-Pip) dipeptide amino-aldehyde from a NRPS module that upon reverse prenylation, suffers a cascade of cyclization, dehydration, tautomerization and intramolecular Diels-Alder cycloaddition to construct the monooxopiperazine bicyclo[2.2.2]diazaoctane ring system common to this family. Through the use of total chemical synthesis of isotopically labeled intermediate metabolites, genome mining, biosynthetic gene cluster identification and functional expression of biosynthetic enzymes, key features of the biosynthetic pathways to these complex secondary metabolites will be experimentally elucidated. In a multi-PI relationship and sub-award with Prof. David Sherman's laboratory (University of Michigan), we are actively engaged in the high-resolution elucidation of the entire biosynthetic pathway to these biomedically significant alkaloids. II. Stephacidins, Notoamides, Waikialoids and Brevianamides: The Dioxopiperazines. The dioxopiperazine family of bicyclo[2.2.2]diazaoctane alkaloids are constructed by a net oxidative transformation of a fully prenylated dioxopiperazine substrate. We have discovered that two orthologous species of Aspergillus produce the opposite enantiomers of Stephacidn A and Notoamide B. This fascinating enantiodivergent biogenesis will be further evaluated using bioinformatics analysis by a new collaborator, Prof. Martin Kreitman, a renowned evolutionary geneticist, to determine the evolutionary mechanisms that resulted in this rare production of opposite enantiomers of these complex alkaloids. III. Citrinadins, Citrinalins, Cyclopiamines and PF1270 Alkaloids: The Decarbonylated Alakloids. As a natural out-growth of our work on the Paraherquamide family of prenylated indole alkaloids, we propose to initiate a new project to study the total synthesis and biosynthesis of these structurally related alkaloids that appear to have arisen biogenetically from the reductive decarbonylation of bicyclo[2.2.2]diazaoctane progenitors. Here also, we shall deploy the powerful synergies of total synthesis, whole genome sequencing, bioinformatics analysis, genome mining and functional expression of biosynthetic enzymes to fully elucidate the corresponding biosynthetic pathways to all three families of alkaloids. In all three sub-projects, total synthesis of the natural products and isotopically-labeled biosynthetic intermediates and probe molecules will be utilized to confirm pathway transformations. New chemical entities generated in this program, either from the biological sources or through chemical synthesis, will be extensively screened and evaluated for biological activities at the Univ. of Michigan Center for Chemical Genomics, the National Human Genome Research Institute, and to Prof. Sachiko Tsukamoto (Japan) for analysis of biological activity using a series of biochemical and cell-based assays relevant to cancer and parasitic disease targets. Additional collaborators include: Prof. Sachiko Tsukamoto, Kumamoto University, Japan; Prof. Jens Frisvad, Technical University, Denmark; Prof. Martin Kreitman, University of Chicago; and Prof. Janet Smith, University of Michigan.

描述（由申请人提供）：本修订后的资助申请侧重于实验研究，以阐明三种生物遗传学相关的天然产物家族的生物合成：(1)Paraherquamides, asperparparines, Malbranchemaides， marmartines和Chrysogenamide (monooxopiperazines)；(2) Stephacidins、Notoamides、Waikialoids和Brevianamides（二氧哌嗪类）；(3)柑桔苷类、柑桔苷类、环胺类和PF1270类生物碱（脱碳生物碱）。目标是利用生物合成酶的全合成、全基因组测序、生物信息学分析、基因组挖掘、功能表达和x射线晶体学的强大协同作用，充分阐明所有三种生物碱家族的相应生物合成途径。1 .对苯二甲胺类、阿斯帕帕林类、丙烯酸类、马马丁内斯类和金曲根酰胺类：单哌嗪类。在我们的实验室中已经阐明了具有争议的证据，表明这类生物碱是由一种罕见的生物合成分子内[4+2]环加成反应构建的，该反应是由NRPS模块还原释放Trp-Pro（或Trp-Pip）二肽氨基醛引起的，在反向戊烯酰化后，遭受级联环化，脱水，通过互变异构和分子内diols - alder环加成，构建了该家族共有的单哌嗪双环[2.2.2]重氮辛烷环体系。通过同位素标记的中间代谢物的全化学合成、基因组挖掘、生物合成基因簇鉴定和生物合成酶的功能表达，实验将阐明这些复杂次生代谢物的生物合成途径的关键特征。在与David Sherman教授的实验室（密歇根大学）的多个pi关系和分奖中，我们积极参与这些生物医学上重要的生物碱的整个生物合成途径的高分辨率阐明。2。石蒜苷类、诺托酰胺类、怀基烷类和brevianamide类：二氧哌嗪类。双环[2.2.2]重氮辛烷生物碱的二氧哌嗪家族是由一个完全的净氧化转化而构建的