Precision H2O2 Management for Pyropia haitanensis Sporulation: Transcriptome-Driven Identification of Peroxidases

To regulate spore formation and induction in Pyropia haitensis , this study investigates potential peroxidase targets. The spore-formation induction techniques used today are highly variable and challenging to control precisely on a large scale. To increase the amount of H ₂ O ₂ , a reactive oxygen species (ROS) known to induce asexual reproduction in Pyropia haitensis and closely related species, this study suggests using molecular precision by inhibiting peroxidases. To identify these critical peroxidases, transcriptomics data from various stages of Pyropia haitensis ’ life cycle were examined. Differential expression analysis and functional annotation, utilizing NCBI BLAST and InterProScan, were employed as the methods. Finally, ligand binding affinities of annotated peroxidase sequences were predicted using GPsite analysis. The findings indicate that some peroxidase gene sequences exhibit stage-specific expression patterns; 18469 and 19494 are identified as potential targets for inhibition due to their decreased expression during the spore formation stage. Numerous peroxidase types, including haloperoxidases and classical haem-dependent peroxidases, were identified by annotation. The majority of the identified peroxidases exhibited high haem binding, as determined by GPsite analysis. High binding to Ca ²⁺ , Mg ²⁺ , and Mn ²⁺ ions, a property of catalases, was demonstrated for specific sequences. Furthermore, peroxidases with high binding affinity to Ca ²⁺ , Mg ²⁺ , and Mn ²⁺ ions lacked annotation related to thylakoid localisation, which made them desirable targets whose inhibition could prevent damage to photosystem II. This study revealed several peroxidases with potential for additional investigation and modification to enhance spore production. This opens the door for future large-scale ligand-based screening using proteo-chemometrics with AlphaFold and in vitro validation with full factorial designs, to maximise on-demand spore production on a large scale.