用激光镊子拉曼光谱法优化红法夫酵母生产虾青素的条件。首先, 将红法夫酵母细胞拉曼光谱与虾青素标准品溶液拉曼光谱进行对比, 找出定量虾青素的拉曼特征峰; 然后对照红法夫酵母生长曲线与不同时间点红法夫酵母细胞内虾青素含量曲线, 进行发酵时间的优化; 最后将红法夫酵母分别用不同氮源、碳源培养基培养, 对比其细胞的虾青素拉曼特征峰强度, 最终优化培养基。由实验可得, 适宜定量虾青素的拉曼特征峰是1520 cm-1峰; 最佳发酵时间为72 h; 最佳优化培养基氮源含硫酸铵4 g/L+硝酸钾4 g/L, 碳源含蔗糖60 g/L。从上述结果可知, 用激光镊子拉曼光谱法对红法夫酵母合成虾青素的条件进行优化, 充分发挥了其操作简便、耗时短、样品用量少、对样品无损伤等优势, 使所得结果更准确可信。因此, 激光镊子拉曼光谱法是红法夫酵母细胞合成虾青素条件优化的理想选择。

In this paper, laser tweezers Raman spectroscopy (LTRS)is used for optimizing fermentation condition of astaxanthin in Phaffia Rhodozyma. First, For identifying the characteristic peak of astaxanthin we compare the Raman spectra of Phaffia Rhodozyma cells with that of astaxanthin standard solution; Then we contradistinguish growth curve of Phaffia Rhodozyma from content curve of astaxanthin in Phaffia Rhodozyma cells in different fermentation time for optimization fermentation time; Finally, we culture Phaffia Rhodozyma cells in different carbon medium or nitrogen medium, and compare the Raman spectra of cells in different medium for the medium optimization. The results are as follows: The peak at 1520 cm-1 is an appropriate characteristic raman peak for astaxanthin quantitation; The optimal fermentation time is 72 h; The optimal medium contain sucrose 60 g/L for carbon source and ammonium sulfate 4 g/L, potassium nitrate 4 g/L for nitrogen source. The above results suggest LTRS when used for optimizing fermentation condition of astaxanthin in Phaffia Rhodozyma possess advantages of simple operation, short time, less sample consumption, no damage to the sample and so on. It makes experimental data more accurate and reliable. Therefore LTRS is an efficient approach for optimizing fermentation condition of astaxanthin in Phaffia Rhodozyma.