Contribution
Our team contributed to the iGEM and synthetic biology community by building an integrated framework for thermophilic polyamine discovery, characterization, and application, combining methodological, genetic, and analytical innovations.
1. Thermophilic Polyamine Gene Resource Development
We established a metagenomic mining and annotation pipeline to identify key polyamine biosynthetic genes from high-temperature microbiomes. A curated database was constructed, covering six core enzyme families involved in polyamine metabolism --- including arginine decarboxylase, ornithine decarboxylase, SAM decarboxylase, spermidine synthase, aspartate kinase, and aspartate semialdehyde dehydrogenase. This database contains over 650 validated thermophilic sequences, providing a valuable genetic resource for future research on enzyme evolution, pathway optimization, and polyamine engineering.
2. Multi-Omics Profiling of Heat Stress Response
We conducted a comprehensive multi-omics analysis (transcriptome, proteome, and metabolome) of representative Bacillus strains cultivated at 37 °C, 45 °C, and 55 °C. The results revealed coordinated thermal adaptation mechanisms involving metabolic reprogramming, protein homeostasis, and membrane regulation, with consistent upregulation of polyamine biosynthetic genes and precursors. This dataset provides a systematic reference map for studying thermophilic stress adaptation and designing robust microbial chassis for high-temperature biosynthesis.
3. Thermotolerant Chassis Candidate Collection
Through selective aerobic and anaerobic culturing across temperature gradients, we successfully isolated and purified 64 thermotolerant strains spanning 25 species and 9 genera, primarily within the Bacillus and Anoxybacillus lineages. Among these, six representative strains were sequenced and annotated, confirming the presence of multiple polyamine biosynthetic pathways.This strain library establishes a living resource for constructing thermophilic production chassis and for future comparative genomic studies on heat adaptation and polyamine metabolism.
4. Integration into a Thermophilic Synthetic Biology Pipeline
All experimental findings were integrated into a modular synthetic biology framework, linking metagenomic discovery, strain isolation, omics characterization, and analytical validation.This unified pipeline empowers downstream iGEM teams and the broader synthetic biology community to replicate, extend, or repurpose thermophilic biosynthetic systems for the sustainable production of polyamines and other bioactive compounds.