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Identifying Antibiotic Molecules in Ceanothus leucodermis and Quantifying Their Antibacterial Activity with a Novel, Simulation-Aided Method

ISEF · 2019 Microbiology

Feng, Daniel · University High School

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Overview

Antibiotic-resistant bacteria have rendered many modern antibiotics ineffective. Plants, however, have traditionally provided many natural medicines. In a previous project, I discovered that the Native American herb Ishwish (Ceanothus leucodermis) showed antibacterial properties. This year, I purified and identified the antibiotics in Ishwish and quantified their antibacterial strength using a novel, simulation-aided method. To determine the optimal method to extract molecules from plant mass, I created several liquid extracts under varying conditions. Antibacterial strength was tested using disk diffusion assays, and an ethanolic extraction of Ishwish stems was found to be the most effective. I purified this extract with C8 and silica gel chromatography columns and evaluated the effectiveness of my purification by using a low-resource method that I developed. My method combines disk diffusion assay data with a diffusion simulation that I wrote in Python. This allowed me to find the MICs (Minimum Inhibitory Concentrations) of my active fractions despite the small amounts available. The purification was found to be successful. Finally, I applied my most purified fractions to LC-MS analysis to identify the antibacterial compounds. After comparing my LC-MS data to molecular data in the database METLIN, I determined that the active compounds were polymers of the antibiotic catechin. I successfully purified and identified the antibiotics in Ishwish and demonstrated a real-life application of my computer simulation to determine MICs. This project provides a scientific basis for the Native Americans’ use of Ishwish, and the strategies developed here can be applied to examine other Native American herbs while using limited amounts of plant material.

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Awards (1)

  • Second Award of $1,500 $1,500

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Source: Regeneron International Science and Engineering Fair

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