Functional and Genetic Analysis of an Esterase Gene Involved in Synthesis of Ergot Alkaloids

Kelcie N. Britton*, Chey R. Steen*, Jessi K. Sampson, and Daniel G. Panaccione
Division of Plant and Soil Sciences, West Virginia University, Morgantown, WV 25606

Presentation Category: Science & Technology (Oral Presentation)

Student’s Major: Immunology and Medical Microbiology

Ergot alkaloids represent a broad class of nitrogenous metabolites produced across a range of fungal species and have been applied as effective pharmaceutical treatments for dementia, migraines, and hyperprolactinemia. Some pathways have been studied both biochemically and genetically, but critical steps in the synthesis of lysergic acid amides remain elusive. Many of the pharmaceutically relevant ergot alkaloids are semi-synthetically derived from lysergic acid amides, making these gaps significant. Lysergic acid ⍺-hydroxyethylamide (LAH) is the main lysergic acid amide produced by the fungus Metarhizium brunneum. We hypothesize a gene named easP encodes an esterase involved in the final step of LAH biosynthesis. To test this hypothesis, a CRISPR mutant was engineered for easP and checked using PCR and DNA sequencing. The quantity of LAH, as measured by high performance liquid chromatography in each strain, was normalized showing that the concentration of LAH in CRISPR-derived mutants was reduced by ~50% relative to that in the wild-type. These data indicate that EasP plays a significant role in producing LAH and that an additional esterase is also involved.

Funding: NIH, 2R15GM114774-02

Program/mechanism supporting research/creative efforts: : a WVU 497-level course