The field of study on extracellular vesicles (EVs) has recently focused on probiotic bacteria, due to their ability to exhibit a wide variety of biological activity. Therefore, this study aims to evaluate the effect of nutrient modulation during Lacticaseibacillus paracasei cultivation on EVs production and bioactivity. EVs were isolated by ultrafiltration method and were characterized using NTA and cryo-TEM. An in vitro evaluation was used to assess the bioactivities, antioxidant properties, anti-inflammatory potential, and ability to inhibit α-glucosidase activity. The results showed that the 2 LPEV exhibited the characteristics of vesicles with size of 96.04–100.29 nm. Cultivation in 50% MRS increased the number of particles (5.70 × 10⁹ particles/mL) compared with 100% MRS (4.46 × 10⁹ particles/mL). In addition, LPEV exhibited moderate antioxidant activity (15.5–34.6%), strong anti-inflammatory activity (90.6–99.2%), and substantial α-glucosidase inhibitory activity (75.5–79.7%). The reducing nutrients increased EV production without changing their physical and chemical properties or biological activity. This suggest that changing nutrients is an easy way to increase L. paracasei EV production and use LPEVs in the future for biomedical studies.
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