(P26-032-25) Comparative Peptidomics-Based Identification of Novel Bioactive Peptides From Probiotics Digestion of Green Tea Biomass

Objectives: Bioactive peptides (BPs) are renowned for their diverse bioactivities, high stability, and low toxicity, making them valuable in cosmetics, pharmaceuticals, and the food industries. The sequence and functionality of BPs depend on protein cleavage sites, emphasizing the importance of hydrolysis method. This study aimed to evaluate functional potential and industrial scalability of BPs from green tea (GT) biomass.

Methods: GT biomass (40% crude proteins) was hydrolyzed enzymatically using trypsin and alcalase, and through microbial fermentation under aerobic and anaerobic conditions with Lactiplantibacillus plantarum APsulloc 331261. Low molecular weight peptides were purified via ultrafiltration and size-exclusion chromatography, and sequences were identified using LC-MS². Bioactive potential was assessed in silico using the BIOPEP database. Media was optimized by determining essential and non-essential amino acids (AAs) using in silico analysis and single omission, and by determining concentrations of media components using RSM.

Results: Hydrolysis of GT biomass under varying conditions resulted in distinct peptide portfolios and cleavage patterns. Anaerobic fermentation yielded 310 GT-derived peptides and 26 microbe-derived peptides. L. plantarum preferentially cleaved between hydrophobic and polar residues, differing from enzymatic hydrolysis patterns. Using a BP database, we predicted DPP-4 and ACE inhibitory activities of these peptides. Growth media optimization involved supplementing essential AAs (S, D, E) while omitting non-essential AAs (W, K, N, R, P), with further optimization of AAs, glucose, and GT biomass concentrations.

Conclusions: The study provides a framework for valorizing GT biomass to produce BPs via microbial fermentation, highlighting the potential of GT biomass as a sustainable protein resource for various applications.

Funding Sources: This research was supported by AMOREPACIFIC, Bio & Medical Technology Development Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Science and ICT (MSIT) (RS-2021-NR056579), and a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (RS-2023-KH141436).