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.

项目概要天然产物作为药物或药物化合物的先导物在制药行业中发挥着至关重要的作用。这些分子通过次级代谢产生，影响毒力、共生和生存非蛋白原性氧乙烯基甘氨酸是一类有趣的次级氨基酸。此类代谢物的特征是存在乙烯基氧部分，例如 4-(2’-)。氨基乙氧基）乙烯基甘氨酸 (AVG) 已知可通过抑制吡哆醛 5'- 来阻止植物发芽磷酸盐 (PLP) 依赖性酶 1-氨基环丙烷-1-羧酸 (ACC) 合酶。2 含乙烯基醚的氧乙烯甘氨酸非蛋白氨基酸（包括 AVG）的生物合成经过探索，具有替代功能组的其他家庭成员的形成仍然存在据报道，4-甲酰氨基氧基乙烯基甘氨酸（FVG）是一种含有 N-O 键的氧基乙烯基甘氨酸。具有除草和杀菌双重活性。3,4假单胞菌中的gvg生物合成基因簇荧光菌株 WH6 负责 FVG 和相关化合物的生物合成胍基氧基乙烯基甘氨酸 (GOVG) 和氨基氧基乙烯基甘氨酸 (AOVG) 的生物合成机制仍不清楚。5,6 在本提案中，我将确定负责形成的生物合成途径 FVG、GOVG 和 AOVG 我将结合喂养实验和体外生化测定来进行。确定这三种生物合成途径中每种酶的底物和产物非蛋白氨基酸。乙烯基烷氧基胺在氨基酸中不常见，因此我将对其进行表征。这些研究将确定负责 N-O 键构建的酶并确定其化学机制。促进类似的氧乙烯甘氨酸非蛋白氨基酸的生物信息学发现。负责 N-O 键形成的铁依赖性酶的表征将有助于其在扩大氧化生物催化剂的合成范围。