A Study on the Extraction, Fermentation Condition Optimization, and Antioxidant Activity Assessment of Polysaccharides Derived from Kluyveromyces marxianus

<i>Kluyveromyces marxianus</i> exhibits advantages such as strong thermotolerance, rapid growth rate, and high safety, making it an excellent host cell for the production of bio-based products. In this study, two exopolysaccharides, KE1 and KE2, were isolated from the fermentation broth of the strain <i>K. marxianus</i> KM-502, and their hydroxyl radical scavenging, DPPH scavenging, and Fe²⁺-reducing activities were evaluated. In vitro antioxidant assays demonstrated that exopolysaccharide KE2 exhibited superior antioxidant activity compared to KE1. The fermentation conditions were optimized using single-factor experiments followed by response surface methodology (RSM). The optimized fermentation process revealed that the most suitable fermentation medium consisted of 8% sucrose, 1.99% peptone, and 0.13% CaCl₂, while the optimal fermentation conditions were a medium volume of 74 mL in a 300 mL flask, pH 6.7, an inoculum size of 1.99%, a temperature of 30 °C, a shaking speed of 160 r/min, and a cultivation time of 96 h. After optimizing the fermentation conditions of <i>K. marxianus</i> KM-502, the exopolysaccharide (EPS) yield reached 5842.42 mg/L, representing a 22.77-fold increase compared to the yield before optimization. In summary, this study isolated exopolysaccharides KE1 and KE2 from <i>K. marxianus</i> KM-502. These exopolysaccharides demonstrated significant antioxidant activities, and the fermentation conditions for exopolysaccharide production were optimized. The findings of this study will facilitate the further development and utilization of exopolysaccharides from <i>K. marxianus</i>.