Department of Food Science and Biotechnology, Gachon University, Seongnam 13120, Korea.
Food Nutrition Major, School of Food, Chungkang College of Cultural Industries, Icheon 17390, Korea.
Molecules. 2018 Aug 23;23(9):2118. 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.
利用中心组合设计的响应面法优化 CCK940 生产低聚糖，以培养温度、蔗糖和麦芽糖浓度为主要因素。预测生产低聚糖的最佳条件为培养温度 30°C、蔗糖浓度 9.6%（/）和麦芽糖浓度 7.4%（/）。利用这些最佳条件，使用发酵罐培养 CCK940 生产低聚糖，得到的聚合度大于 4 的低聚糖通过 Bio-gel P2 凝胶渗透柱色谱法纯化，然后冻干。当巨噬细胞用浓度为 0.1⁻10 mg/mL 的纯化低聚糖处理时，巨噬细胞没有细胞毒性。然而，一氧化氮的产生水平与 1 μg/mL 脂多糖处理后的水平相似。用低聚糖处理后，肿瘤坏死因子-α、白细胞介素-1β、白细胞介素-6 和诱导型一氧化氮合酶的 mRNA 表达水平均呈剂量依赖性增加。这些数据表明，CCK940 产生的低聚糖可用作巨噬细胞的免疫增强剂。
