胶质芽孢杆菌胞外多糖单糖组成的研究

Research on monosaccharide composition of polysaccharide produced by Bacillus mucilaginosus

中文关键词:  胶质芽孢杆菌PM13  多糖  单糖组成；气相色谱法

英文关键词:Bacillus mucilaginosus PM13 strain  polysaccharide  monosaccharide composition  gas chromatography

中文摘要:

      目的 对胶质芽孢杆菌PM13菌株的胞外多糖的单糖组成进行研究。方法 取培养48 h的斜面菌体, 用10倍体积的蒸馏水稀释, 稀释液6000 r/min离心30 min后除去沉淀, 上清液加4倍体积的95%的乙醇进行沉淀, 并于6000 r/min离心30 min得沉淀物, 沉淀物烘干得到多糖样品。采用苯酚-硫酸法测定多糖样品中含糖量。采用气相色谱法分析其单糖组成, 色谱条件为: 进样口温度为250 ℃, 分流比为40:1, FID检测器温度为250 ℃, VF-5HT熔融石英毛细柱, 柱温为160 ℃, 8 ℃/min升至200 ℃, 200 ℃保持5 min, 载气为高纯氮气, 1.6 mL/min。结果 PM13菌株胞外多糖的含糖量为90.98%, 其单糖组成为葡萄糖、半乳糖、甘露糖和木糖, 比例为5.57:3.08:4.04:1。结论 本文建立了用气相色谱法检测胶质芽孢杆菌PM13菌株胞外多糖的单糖组成的方法, 确定了胶质芽孢杆菌胞外多糖的单糖组成及相应比例。

英文摘要:

      Objective To study the monosaccharide composition of the exopolysaccharides produced by Bacillus mucilaginosus PM13. Method Bacillus mucilaginosus PM13 was cultured on agar slant for 48 h, diluted by 10 times volume of distilled water, centrifuged at 6000 r/min for 30 min. The supernatant was collected and mixed with 4 times volume of 95% ethanol, and centrifuged at 6000 r/min for 30 min to obtain pre-cipitate. The precipitate was dried to obtain the polysaccharide sample. The content of sugar was determined by phenol sulfuric acid method. The monosaccharide composition was analyzed by gas chromatography. The chromatographic condition was as follows: injector temperature 250 ℃, spilt ratio 40:1, FID detector temperature 250 ℃, VF-5HT melted silica capillary column. The initial column temperature was 160 ℃, then increased to 200 ℃ with the rate of 8 ℃/min and kept this temperature for 5 min. The carrier gas was high purity nitrogen gas with the speed of 1.6 mL/min. Results The sugar content of sample was 90.98% on average. The results of the gas chromatography analysis showed that the polysaccharide produced by PM13 strain was mainly composed of glucose, galactose, mannose, and xylose with the ratio of 5.57:3.08:4.04:1. Conclusion The detection method of the monosaccharide composition of the exopolysaccharides from Bacillus mucilaginosus PM13 by gas chromatography is established and the monosaccharide composition and the rate are obtained.