Biosynthesis of Several Oxyvinylglycine Nonproteinogenic Amino Acids Bearing Unusual Alkoxyamine Bonds

几种带有异常烷氧基胺键的氧乙烯甘氨酸非蛋白氨基酸的生物合成

基本信息

批准号：
10752052
负责人：
Joshua Ray Turek-Herman
金额：
$ 6.87万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10752052
关键词：
Agrochemicals Amino Acids Anabolism Anti-Bacterial Agents Antibiotics Attenuated Bacteria Binding Biochemical Bioinformatics Biological Assay Breathing C-terminal Cells Chemicals Dependence Drug Compounding Drug Industry Enzyme Inhibition Enzymes Ethers Ethylenes Exhibits Family member Gene Cluster Genetic Germination Goals Guanidines Heme Herbicides Homoserine Imino Acids In Vitro Industrialization Iron Medicine Metabolism Metals Methods N hydroxylation N-terminal Natural Compound Natural Products Nucleosides Oxygen Oxygenases Pathway interactions Pharmaceutical Preparations Pharmacologic Substance Plant Growth Regulators Plants Property Pseudomonas fluorescens Pyridoxal Phosphate Reaction Recombinants Reporting Role Source Streptozocin Structure Symbiosis Synthesis Chemistry System Virulence bactericide carboxylate carboxylation chemotherapy cofactor enol enzyme mechanism experimental study feeding functional group member metalloenzyme microorganism secondary metabolite small molecule small molecule inhibitor stable isotope unnatural amino acids vinyl ether

项目摘要

Project Summary Natural products serve crucial roles in the pharmaceutical industry as drugs or leads for drug compounds. Generated through secondary metabolism, these molecules impact the virulence, symbiosis, and survival of microorganisms. The nonproteinogenic oxyvinylglycine amino acids are an interesting class of secondary metabolites characterized by the presence of a vinyl oxygen moiety. Members of this class, such as 4-(2’- aminoethoxy)vinylglycine (AVG), are known to arrest germination in plants by inhibiting the pyridoxal 5’- phosphate (PLP)-dependent enzyme 1-aminocyclopropane-1-carboxylate (ACC) synthase.2 While the biosynthesis of vinyl ether-bearing oxyvinylglycine nonproteinogenic amino acids including AVG has been explored, the formation of other members of the family with alternative functional groups remains undetermined. 4-Formylaminooxyvinylglycine (FVG), an oxyvinylglycine containing an N–O bond, was reported to have duel herbicidal and bactericidal activity.3,4 The gvg biosynthetic gene cluster in Pseudomonas fluorescens strain WH6 is responsible for the biosynthesis of FVG and the related compounds guanidinooxyvinylglycine (GOVG) and aminooxyvinylglycine (AOVG) though the exact biosynthetic mechanism remains unclear.5,6 In this proposal I will determine the biosynthetic pathways responsible for the formation of FVG, GOVG, and AOVG. I will use a combination of feeding experiments and in vitro biochemical assays to determine the substrate and product of each enzyme along the biosynthetic pathway of these three nonproteinogenic amino acids. The vinyl alkoxyamine is unusual in amino acids, therefore I will characterize the enzyme responsible for N–O bond construction and determine its chemical mechanism. These studies will facilitate the bioinformatic discovery of similar oxyvinylglycine nonproteinogenic amino acids. Moreover, characterization of the iron-dependent enzyme responsible for N–O bond formation will facilitate its use in a synthetic context to expand the repertoire of oxidative biocatalysts.

项目摘要天然产品在制药行业中起着至关重要的作用，作为药物或药物化合物的铅。这些分子通过继发代谢产生，影响病毒，共生和生存微生物。非蛋白质生成的氧乙烯基甘氨酸氨基酸是一类有趣的次级代谢产物的特征是存在乙烯基氧部分。该课的成员，例如4-（2'-- 已知通过抑制吡啶多5'-- 磷酸盐（PLP）依赖性酶1-氨基丙烷-1-羧酸盐（ACC）合酶2 乙烯基乙醚含氧乙烯基乙烯基乙烯基非蛋白质生成氨基酸（包括AVG）的生物合成已是经过探索，仍然存在替代功能组的家庭其他成员的形成不确定。据报道，4-甲基氨基乙烯基甘氨酸（FVG）是含有N – O键的氧乙烯基甘氨酸要育成纯种和杀菌活性。3,4假单胞菌中的GVG生物合成基因簇荧光菌株WH6负责FVG的生物合成和相关化合物的生物合成瓜尼迪二乙烯基甘氨酸（GOVG）和氨基乙烯基甘氨酸（AOVG）虽然确切的生物合成机制仍然不清楚。5,6在该提案中，我将确定负责形成的生物合成途径 FVG，Govg和AOVG。我将使用喂养实验和体外生化测定的组合确定沿这三个酶的生物合成途径的每个酶的底物和产物非蛋白质生成氨基酸。乙烯基烷氧胺在氨基酸中不寻常，因此我将表征负责N – O键构建并确定其化学机制的酶。这些研究会促进生物信息学发现相似的氧乙烯基甘氨酸非蛋白质生成氨基酸。而且，负责N – O键形成的铁依赖性酶的表征将促进其在A中的使用合成环境扩大氧化生物催化剂的曲目。