Overview
This project explored using methane-consuming bacteria (Methylomicrobium alcaliphilum) and engineered E. coli as biocatalysts for producing shikimate and muconic acid — key precursors in the biosynthesis of nylon — as a sustainable alternative to petroleum-based chemical production. The work was conducted at the UNT BioDiscovery Institute under Dr. Calvin Henard and included a science communication component adapting research findings for a young audience.
Methods / Approach
- Designed a heterologous shikimate biosynthesis pathway by identifying and selecting enzyme-coding genes for introduction into a E. coli plasmid construct.
- Performed Gibson Assembly to construct the multi-insert plasmid and validated assembly by sequencing and colony PCR.
- Conducted qRT-PCR to assess transcript levels of inserted pathway genes under methane-feeding conditions in the bioreactor system.
- Adapted the Henard Lab’s published research (Muconic acid production from methane using rationally-engineered methanotrophic biocatalysts) into a plain-language article for Frontiers for Young Minds — a peer-reviewed journal targeting readers aged 8–15.
Key Results
- Successfully assembled and verified the target plasmid construct containing the shikimate pathway insert.
- qRT-PCR confirmed transcription of inserted genes under experimental growth conditions.
- Frontiers for Young Minds adaptation submitted, translating enzyme engineering and green chemistry concepts for a lay audience.
- Awarded $500 in internal research funding for project proposal and preliminary results.
- Gibson Assembly — multi-fragment cloning for pathway construction
- qRT-PCR — transcript-level verification of inserted genes
- Bioreactor Design — scalable methane-fed bioreactor for muconic acid / nylon precursor production
- Benchling (LIMS) — sequence design, annotation, and lab notebook
- Anaerobic Culture (Coy chamber) — methanotroph cultivation under controlled atmosphere
- Science Communication — Frontiers for Young Minds manuscript preparation