Strategic reprogramming of the ergot alkaloid pathway

麦角生物碱途径的战略重编程

• 批准号: 8878600
• 负责人: Daniel G. Panaccione
• 金额: $ 33.29万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2018-09-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8878600
• 关键词: Acids; Address; Affect; Alkaloids; Alleles; Alzheimer' s Disease; Anabolism; Aspergillus fumigatus; Biochemical; Candidate Disease Gene; Chemistry; Claviceps; Complex; Cytochrome P450; Dementia; Development; Disease; Engineering; Ergot Alkaloids; Family; Future; Gene Cluster; Genes; Genetic; Goals; Health; Healthcare; Human; Hyperprolactinemia; Individual; Isomerase; Knock-out; Methods; Migraine; Mixed Function Oxygenases; Modification; Molecular; Mutation; Nature; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Oxidoreductase; Parkinson Disease; Pathway interactions; Pharmacologic Substance; Production; Relative (related person); Role; Senile dementia; Source; System; Variant; Work; base; clinically relevant; experience; fungus; graduate student; improved; in vivo; mutant; novel; oxidation; pi bond; premature; public health relevance; undergraduate student

 DESCRIPTION (provided by applicant): Ergot alkaloids improve human health as powerful and versatile pharmaceuticals for treatment of multiple conditions including senile dementia, Alzheimer's disease, Parkinson's disease, migraines, hyperprolactinemia, and type 2 diabetes. Different types of ergot alkaloids are produced by distantly related groups of fungi. Certain species of Claviceps and Epichloë produce ergot alkaloids derived from lysergic acid, and these have powerful pharmacological activity. Two other species of Claviceps make dihydroergot alkaloids, in which slightly different chemistry results in different but equally powerful activitis. A third group of fungi, including Aspergillus fumigatus, makes a family of ergot alkaloids that is no derived from lysergic acid or dihydrolysergic acid and is not used clinically; however, A. fumigatus grows faster, is more readily manipulated, and has greater potential for genetically improving ergot alkaloids. All these fungi share common early steps in their ergot alkaloid pathways before diverging to make their unique end products. By blocking the A. fumigatus pathway at a specific point and adding two selected genes from a lysergic acid producer, A. fumigatus has been modified to produce lysergic acid. The long-term goals of the proposed project are to modify and expand the A. fumigatus expression system to answer basic questions about the origins of lysergic acid and related compounds. In addition to answering basic genetic questions, the work will have practical healthcare-related implications, because modified fungal strains will produce important ergot alkaloids that are rare in nature but are pharmaceutically relevant compounds. Specific aims are to (1) Clarify steps in the production of lysergic acid and elucidate the biosynthetic origins of alternate ergot alkaloids paspalic acid and lysergol, and (2) Determine the biosynthetic origin of dihydroergot alkaloids. Individual genes or combinations of two genes will be isolated from fungi that make these unique ergot alkaloids and expressed in A. fumigatus modified to produce one of two different substrate ergot alkaloids. The A. fumigatus strains transformed with these genes will be analyzed by molecular and biochemical methods to determine how expression of the candidate genes has affected the fungus's ergot alkaloid profile. Results of the proposed project will reveal roles of specific genes and elucidate origins of several important ergot alkaloids. The project will produce strains of A. fumigatus that produce molecules with pharmaceutical significance. Additional benefits include meaningful experiences for graduate and undergraduate students and further development of a platform for modification and improvement of additional or novel ergot alkaloids.

 描述（由适用提供）：麦角生物碱改善了人类健康，作为功能强大且多功能的药物，用于治疗多种疾病，包括老年痴呆症，阿尔茨海默氏病，帕金森氏病，偏头痛，高脂蛋白血症和2型糖尿病。遥远相关的真菌组产生了不同类型的麦角生物碱。某些种类的claviceps和Epichloë产生了源自脂肪酸的麦角生物碱，并且具有强大的药物活性。另外两种种类的claviepeps产生了二氢豆激素生物碱，其中略有不同的化学作用会导致不同但同样强大的活性。第三组真菌，包括曲霉菌，使麦角生物碱家族不可能源自脂肪酸或二氢溶质酸，并且在临床上未使用。然而，烟曲霉的生长速度更快，更容易受到操纵，并且具有更大的基因改善麦角生物碱的潜力。所有这些真菌在分歧以制造其独特的最终产品之前，在其麦角生物碱途径中共享共同的早期步骤。通过在特定点阻止烟曲霉途径并添加来自脂肪酸生产者的两个选定基因，已对富马图斯曲霉进行了修改以产生脂肪酸。拟议项目的长期目标是修改和扩展烟曲霉的表达系统，以回答有关脂肪酸及相关化合物起源的基本问题。除了回答基本的遗传问题外，这项工作还将具有与医疗保健相关的实际含义，因为经过修改的真菌菌株还会产生重要的麦角生物碱，这些生物碱本质上很少，但具有物理上相关的化合物。具体目的是（1）阐明产生脂肪酸的步骤，并阐明替代性麦角生物碱生物碱的生物合成起源 - paspalic Acid and Lysergol，以及（2） 确定二羟基果仁生物碱的生物合成起源。将从真菌中分离出两个基因的单个基因或组合，这些真菌使这些独特的麦角生物碱并在烟曲霉中表达，以产生两种不同的底物麦角麦尔菌之一。用这些基因转化的烟曲霉菌株将通过分子和生化方法分析，以确定候选基因的表达如何影响真菌的麦角生物碱谱。拟议项目的结果将揭示特定基因的作用并阐明 几种重要的麦角生物碱的起源。该项目将产生烟曲霉的菌株 产生具有药物意义的分子。其他好处包括为研究生和本科生的有意义的经历以及改进和改善其他或新颖的麦角生物碱的平台的进一步发展。