Strategic Reprogramming of the Ergot Alkaloid Pathway

麦角生物碱途径的战略性重编程

• 批准号: 10793120
• 负责人: Daniel G. Panaccione
• 金额: $ 8.64万
• 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 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10793120
• 关键词: Aging; Alzheimer' s Disease; Anabolism; Award; CRISPR/Cas technology; Data; Dementia; Development; Equipment; Ergot Alkaloids; Future; Genes; Goals; Health; High Pressure Liquid Chromatography; Human; Hyperprolactinemia; Learning; Maintenance; Mass Spectrum Analysis; Migraine; Modification; Mutate; Non-Insulin-Dependent Diabetes Mellitus; Parents; Parkinson Disease; Pathway interactions; Pharmacologic Substance; Recombinant DNA; Research; Role; Senile dementia; Statistical Data Interpretation; Structure; System; Testing; Training; Training and Education; Water; data management; design; experience; fungus; gene function; gene product; graduate student; improved; instrument; novel; programs; undergraduate student

7. Project Summary/Abstract 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. Tremendous diversity in structure and activity can be found among the natural and semi-synthetic ergot alkaloids. Small changes in structure may lead to a great changes in activity. The long-term goal of this research program is to understand and ultimately control the biosynthesis of diverse ergot alkaloids by determining the functions of genes and gene products that produce and diversify ergot alkaloids. The requested equipment, a Waters Arc HPLC (high performance liquid chromatography) system, will replace an aging instrument that presents frequent maintenance and data management challenges. The new instrument will enhance efficiency of the research team and improve the quality of data. Working with the new instrument will provide undergraduate and graduate students with relevant training. The instrument will be used to gather data for each of the approaches being used for both specific aims of the parent award, titled “Strategic reprogramming of the ergot alkaloid pathway.” Both specific aims involve modifying fungal ergot alkaloid pathways by CRISPR/Cas9-based approaches or expression of heterologous genes, and then analyzing the altered ergot alkaloid profiles by HPLC and subsequent mass spectrometry approaches. The new HPLC will have an integral role in gathering data for all of the planned experimental approaches. The HPLC will be critical in identifying which candidate strains have been mutated by the recombinant DNA approaches and in identifying the metabolites accumulating in those constructed strains. The HPLC also will be used to collect quantitative data on ergot alkaloid accumulation which will be used in statistical analyses to test hypotheses about gene function. Results of the proposed project will reveal roles of specific genes and provide strains of fungi that produce molecules with pharmaceutical significance. Additional benefits include meaningful experiences for graduate and undergraduate students and further development of platforms for modification and improvement of additional or novel ergot alkaloids.

7. 项目总结/摘要 麦角生物碱作为强大且多功能的药物可改善人类健康，用于治疗多种疾病 包括老年痴呆症、阿尔茨海默病、帕金森病、偏头痛、 高泌乳素血症和 2 型糖尿病在结构和活性上存在巨大的多样性。 在天然和半合成的麦角生物碱中，结构上的微小变化可能会导致很大的变化。 该研究计划的长期目标是了解并最终控制活动的变化。 通过确定基因和基因产物的功能来生物合成多种麦角生物碱 生产和多样化麦角生物碱所需的设备是 Waters Arc HPLC（高性能）。 液相色谱）系统，将取代需要频繁维护和维护的老化仪器 新仪器将提高研究团队的效率和数据管理挑战。 使用新仪器将为本科生和研究生提供数据质量的提高。 受过相关培训的学生将使用该仪器收集每种方法的数据。 用于家长奖的两个具体目标，标题为“麦角生物碱的战略重编程” “这两个具体目标都涉及通过基于 CRISPR/Cas9 的方法修改真菌麦角生物碱途径 异源基因的方法或表达，然后分析麦角生物碱谱 HPLC 和后续的质谱方法 新的 HPLC 将在其中发挥不可或缺的作用。 收集所有计划的实验方法的数据对于识别至关重要。 哪些候选菌株已通过重组 DNA 方法突变并鉴定 HPLC 也将用于定量收集这些构建的菌株中积累的代谢物。 麦角生物碱积累的数据将用于统计分析以检验以下假设 拟议项目的结果将揭示特定基因的作用并提供菌株。 产生具有药学意义的分子的真菌还包括有意义的好处。 研究生和本科生的经验以及进一步开发平台 额外或新型麦角生物碱的修饰和改进。

项目成果

Contribution of a novel gene to lysergic acid amide synthesis in Metarhizium brunneum.

• DOI: 10.1186/s13104-022-06068-2
• 发表时间: 2022-05-18
• 期刊: BMC research notes
• 影响因子: 1.8

Functional analysis of the gene controlling hydroxylation of festuclavine in the ergot alkaloid pathway of Neosartorya fumigata.

• DOI: 10.1007/s00294-016-0591-5
• 发表时间: 2016-11
• 期刊: CURRENT GENETICS
• 影响因子: 2.5
• 作者: Bilovol, Yulia;Panaccione, Daniel G.
• 通讯作者: Panaccione, Daniel G.

Derivation of the multiply-branched ergot alkaloid pathway of fungi.

• DOI: 10.1111/1751-7915.14214
• 发表时间: 2023-04
• 期刊: Microbial biotechnology
• 影响因子: 5.7

Ergot Alkaloids of the Family Clavicipitaceae.

• DOI: 10.1094/phyto-12-16-0435-rvw
• 发表时间: 2017-05
• 期刊: Phytopathology
• 影响因子: 3.2
• 作者: Florea S;Panaccione DG;Schardl CL
• 通讯作者: Schardl CL

Psychoactive plant- and mushroom-associated alkaloids from two behavior modifying cicada pathogens

• DOI: 10.1016/j.funeco.2019.06.002
• 发表时间: 2019-10-01
• 期刊: FUNGAL ECOLOGY
• 影响因子: 2.9
• 作者: Boyce, Greg R.;Gluck-Thaler, Emile;Kasson, Matt T.
• 通讯作者: Kasson, Matt T.