Optimization of Oligosaccharide Production from Leuconostoc lactis Using a Response Surface Methodology and the Immunostimulating Effects of These Oligosaccharides on Macrophage Cells

• Published in: Molecules (Basel, Switzerland) Switzerland), 2018-08, Vol.23 (9), p.2118
• Main Authors: Lee, Sulhee, Park, Gwi-Gun, Jang, Jae-Kweon, Park, Young-Seo
• Format: Article
• Language: English
• Subjects: Acids; Alginates; Alginic acid; Animals; Bacteria; Cell culture; Cell Survival - drug effects; central composite design; Cytotoxicity; immunostimulation; Interleukin-1beta - genetics; Interleukin-6 - genetics; Lactic acid; Lactic acid bacteria; Leuconostoc - metabolism; Leuconostoc lactis; Lipopolysaccharides; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Maltose; Mice; Nitric oxide; Nitric Oxide Synthase Type II - genetics; Nitric Oxide Synthase Type II - metabolism; Nitric-oxide synthase; Nitrogen Oxides - metabolism; oligosaccharide; Oligosaccharides; Oligosaccharides - metabolism; Oligosaccharides - pharmacology; Optimization; RAW 264.7 Cells; RAW264.7; Response surface methodology; RNA, Messenger - genetics; Studies; Sucrose; Sugar; Temperature; Tumor Necrosis Factor-alpha - genetics; Tumor necrosis factor-TNF; Tumor necrosis factor-α
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules23092118

Production of oligosaccharides from CCK940 was optimized using a response surface methodology with a central composite design. Culture temperature and the concentrations of sucrose and maltose were used as the main factors. The predicted optimum conditions for the production of oligosaccharides were a culture temperature of 30 °C, a sucrose concentration of 9.6% ( / ), and a maltose concentration of 7.4% ( / ). Using these optimal conditions, CCK940 was cultured using a fermenter to produce oligosaccharides, and the resulting oligosaccharides with a degree of polymerization greater than 4 were purified by Bio-gel P2 gel permeation column chromatography and then lyophilized. When macrophages were treated with the purified oligosaccharides at concentrations of 0.1⁻10 mg/mL, no cytotoxicity towards the macrophages was observed. However, nitric oxide production levels were similar to those following treatment with 1 μg/mL lipopolysaccharide. The mRNA expression levels of tumor necrosis factor-α, interleukin-1β, interleukin-6, and inducible nitric oxide synthase were all also increased in a dose-dependent manner following treatment with the oligosaccharides. These data suggest that oligosaccharides produced by CCK940 could be used as an immune enhancer of macrophages.