HPGPC柱串联法测定肠膜明串珠菌发酵产物研究

Determination of Leuconostoc mesenteroides fermentation by HPGPC columns in series

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。

Leuconostoc mesenteroides is grown on sucrose medium and synthesises different molecular weight of dextran, the fermentation broth contains sucrose substrate, dextran product, fructose, glucose and other byproducts. Tracking and detecting the variations of sucrose, fructose, and dextran product at the same time,are quite important for calculating the consumption of substrate, the amount of by-products and to explore dextran synthesis kinetic. This article uses single column KS 801, two column TSK G4000 PW and KS 801 connected in series in high performance gel permeation chromatography （HPGPC） system, measuring the molecular weight of different dextran standards, and the fermentation obtained by Leuconostoc mesenteroides. The study showed that column TSK G4000 PW coupled with column KS 801 can separate the samples better. The sucrose concentration and peak area shows good linear relationship in the range of 0.1×10^-3-0.1×10^-2 mol/L, and the RSD values of this method are less than 3.00%. Two columns in series in HPGPC system is an accurate, reliable method which is suitable for the analysis of samples containing small molecule carbohydrates and macromolecules.