Identification of a Gene Involved in Production of Pharmaceutically Important Lysergic Acid Amides

Kelcie N. Britton*, Chey R. Steen, Jessi K. Sampson, and Daniel G. Panaccione

Division of Plant and Soil Sciences, West Virginia University, Morgantown, WV 26506

Presentation Category: Oral-Science & Technology (Oral Presentation #27)

Student’s Major: Immunology and Medical Microbiology

Ergot alkaloids are lysergic acid containing compounds produced by several species of fungi associated with significant human and animal toxicoses. Despite their toxicity, modified and appropriately dosed ergot alkaloid derivatives are effective pharmaceutical treatments for dementia, migraines, and hyperprolactinemia. Biochemical pathways to some ergot alkaloids have been determined, but the final step in the synthesis of lysergic acid amides remains elusive. This gap is significant because many of the pharmaceutically relevant ergot alkaloids are derived from lysergic acid amides. Lysergic acid ⍺-hydroxyethylamide (LAH) is the main ergot alkaloid produced by the fungus Metarhizium brunneum. We hypothesize two genes, named easP and estA, encode esterases involved in the final step of LAH biosynthesis. To test this hypothesis, CRISPR mutants were engineered in a M. brunneum background with easP alone mutated and with both estA and easP mutated. Biochemical analysis of our mutant strains demonstrated the product of easP has a significant role in the production of LAH; where the easP mutant only accumulated half of the LAH measured in non-mutant strains when normalized relative to fungal biomass as estimated by the fungal metabolite ergosterol. Mutation of estA did not affect accumulation of lysergic acid amides, indicating another enzyme is contributing the easP redundant activity. The hypothesized activity of EasP as an esterase is being tested by expressing the protein in E. coli. The phenotype of our CRISPR mutant demonstrates that easP is an integral part of the pathway to LAH.

Funding: Arnold and Mabel Beckman Foundation, NIH

Program/mechanism supporting research/creative efforts: an external internship or other type of program, Arnold and Mabel Beckman Foundation