[P002] Poster

1:00 PM - 1:50 PM JST(4:00 AM - 4:50 AM UTC)

[P01-034] Enzyme Screening and Utilization of Enzyme Complexes for Polysaccharide Degradation from Red Algae

○Jiyoon Baek¹, Yeonseo Nam¹, Jeongeun hyeon¹ (1. Sungshin Women's University (Korea))

Keywords:

Enzyme complex,Galacto-oligosaccharides,Red algal,prebiotics

Red algal polysaccharides, including agar, carrageenan, and porphyran, were enzymatically degraded into galacto-oligosaccharides (GOS), which had significant industrial value as prebiotic ingredients. Therefore, enzymatic conversion of red algal polysaccharides enhanced the efficiency of prebiotic production and promoted the growth of probiotic bacteria. In this study, exo-agarase, endo-agarase, NABH, carrageenase, sulfatase, and porphyranase were employed for polysaccharide degradation. All recombinant enzymes were expressed in Escherichia coli BL21(DE3), were purified, and were confirmed by SDS-PAGE. The degradation activity of each purified enzyme was evaluated using the DNS assay with agar, carrageenan, and porphyran as substrates. To investigate synergistic effects, enzymes were combined in pairs at a 1:1 ratio and were screened for optimal activity. Based on the screening results, an enzyme complex was constructed via PCR-based cloning using the most effective enzyme combination. Protein structures and active sites were analyzed using AI-based structural prediction tools. Based on the predicted structures, site-directed mutagenesis was designed to improve expression levels and catalytic efficiency. In addition, AI-guided in silico modeling was performed to optimize enzyme complex formation and improve structural stability. The resulting enzyme complex is expected to exhibit higher GOS production efficiency compared with individual enzymes. The produced GOS is expected to be utilized for probiotic cultivation, and further studies will explore its application in next-generation synbiotic systems and value-added bioprocess development.