脱脂乳发酵体系中混合乳酸菌的HPLC研究

HPLC study of mixed lactic acid bacteria cultured in skim milk

研究了对脱脂乳发酵体系中Hansen DVS发酵后混合乳酸菌的HPLC分析过程中,不同色谱条件对分离效果的影响。确立了最佳色谱条件:样品上样于Toyopearl TSKgel SuperQ-650c强阴离子交换树脂柱,以浓度为0.02 mol/L哌嗪-HC1缓冲液平衡,洗脱盐浓度为1.0 mol/L的NaC1,洗脱梯度为0～0.75 mol/L的NaCl线性梯度洗脱30 min,再经浓度为0.75～1 mol/L的NaCl线性梯度洗脱10 min以及浓度为1 mol/L的NaC1洗脱10 min,流速定为1 mL/min;并对脱脂乳中不同培养时间的Hansen DVS混合乳酸菌进行了色谱表征。首次对脱脂乳发酵体系中混合乳酸菌的HPLC分析最佳色谱条件的建立,为深入研究发酵过程中乳酸菌混合菌群的动态变化提供了一种快速有效的方法。

The influence of isolation effect of diferent chromatographic conditions of mixed lactic acid bacteria ctflmred in skim milk by high performance ion exchange chromatography was studied.The optimum chromatographic conditions were determined. The sample was applied to the chromatography column packed with Toyopearl SuperQ-650c anion resin, equilibrated with 0.02 mol/L piperazin-hydrochloric acid bufer （pH=7.0）,and elution salt 1 mol/L NaC1, eluted with the gradient of 0-75% NaC1/30 min, then 75%-100% NaCl/10 min followed by 100% NaCl ehited for 10 rain at the flow rate 1 mL/min.Under these conditions,mixed lactic add bacteria were properly isolated.And the mixed lactic add bacteria ctflmred in skim milk for different time were characterized. The establishment of the HPLC analysis on mixed lactic add bacteria cultured in skim milk for the first time helps to investigate the dynamic changes of mixed lactic acid bacteria during fermentation.