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蔗糖经肠膜明串珠菌发酵可生成不同分子量右旋糖酐,发酵液中除含有蔗糖底物及右旋糖酐产物外,还有一定的果糖、葡萄糖等副产物。对发酵液中蔗糖、果糖的含量以及产物右旋糖酐的分子量同时进行跟踪检测,对于计算底物消耗量、产物生成量以及探讨右旋糖酐的合成过程规律有重要意义。分别采用KS 801单柱、TSK G4000 PW与KS 801双柱串联的高效凝胶渗透色谱法(HPGPC),测定不同分子量右旋糖酐标准品,以及肠膜明串珠菌粗酶液发酵蔗糖而得的发酵液。研究表明,TSK G4000 PW与KS 801双柱串联对样品的分离效果更好。蔗糖浓度在0.1×10-3~0.1×10-2mol/L范围内与峰面积呈现良好的线性关系,且该方法的RSD值小于3.00%。表明,双柱串联的高效凝胶渗透色谱法准确、可靠,适用于分析样品中同时含有小分子糖类及大分子物质的研究。
Fermentation of sucrose by S. enteritidis can produce dextran of different molecular weights. In addition to the sucrose substrate and dextran product, there are certain fructose, glucose and other by-products in the fermentation broth. The content of sucrose and fructose in the fermentation broth and the molecular weight of the product dextran are tracked at the same time, which is of great significance for calculating the substrate consumption, product generation and the law of dextran synthesis. High performance gel permeation chromatography (HPGPC) with single-column KS 801, double-column TSK G4000 PW and KS 801 was used to determine the contents of dextran with different molecular weight and fermented sucrose from Leuconostoc mesenteroides . Studies have shown that TSK G4000 PW and KS 801 two-column in-line separation of the sample better. The concentration of sucrose showed a good linear relationship with the peak area in the range of 0.1 × 10-3 ~ 0.1 × 10-2mol / L, and the RSD value of this method was less than 3.00%. The results showed that the high performance gel permeation chromatography with two columns in series was accurate and reliable and suitable for the analysis of samples containing both small sugars and macromolecules.