Purification of Leu. lactis SBC001 Oligosaccharides

Oligosaccharides produced by Leu. lactis SBC001 were purified as described in a previous study [17 (link)]. The culture supernatant was concentrated 10-fold at 60 °C using a rotary evaporator (SB-1200, EYELA, Miyagi, Japan), and the concentrate was loaded onto Bio-gel P2 resin (Bio-Rad Laboratories, Inc., Hercules, CA, USA) packed in a glass Econo-Column (1.5 × 120 cm, Bio-Rad Laboratories, Inc.). The elutes were collected using a fraction collector (Gilson Inc., Middleton, WI, USA) with a fraction volume of 5 mL per tube with a flow rate of 0.5 mL/min. The fractions were analyzed by thin-layer chromatography (TLC), and the oligosaccharide fractions were pooled and freeze-dried using a freeze dryer (SunilEyela, Seongnam, Korea). The freeze-dried powder was dissolved in distilled water at a concentration of 1% (w/v).