Microalgal Molecular Farming for Next-Generation Biopharmaceuticals: Integrating Chloroplast Engineering, Synthetic Biology, Structural Bioinformatics, and Artificial Intelligence

Microalgal molecular farming represents a promising platform for sustainable recombinant biopharmaceutical production. This review integrates advances in chloroplast engineering, synthetic biology, and artificial intelligence with experimental evidence and comparative bioinformatic analyses. Chloroplast genomes from eight representative microalgal species were examined to evaluate genomic organization, phylogenetic relationships, and synteny relevant to genetic engineering. Experimental studies involving recombinant therapeutic proteins, antibodies, and vaccine antigens were reviewed to assess the potential of different microalgal expression systems. Additionally, ten biomedically relevant proteins were comparatively analyzed to characterize structural complexity, receptor-binding interfaces, and evolutionary conservation. Detailed structural analyses of erythropoietin (EPO), interferon-α2b (IFN-α2b), and vascular endothelial growth factor A (VEGF-A) evaluated glycosylation-site accessibility, solvent-accessible surface area, intermolecular interactions, disulfide bonds, and structural conservation. The findings highlight protein-specific requirements for folding, post-translational modifications, and receptor recognition, emphasizing the importance of matching therapeutic protein characteristics with suitable microalgal expression strategies. Overall, this integrated approach provides a framework for rational host selection and the development of next-generation microalgal biofactories for recombinant biopharmaceutical production.