低聚半乳糖对植物乳杆菌发酵乳特性及抗蜡样芽孢杆菌活性的改善
摘要: 为探究低聚半乳糖对植物乳杆菌发酵乳特性及抗菌活性的影响，本文采用单因素法考察影响发酵乳特性的主要因素，并以响应面法优化发酵乳最佳发酵条件；以产肠毒素蜡样芽孢杆菌HN001为指示菌，探究低聚半乳糖的添加对植物乳杆菌ZDY2013发酵乳抑菌活性的改善作用。结果表明：植物乳杆菌能有效利用低聚半乳糖进行体外代谢，并抑制蜡样芽孢杆菌生长；牛奶中添加适量低聚半乳糖能够增加植物乳杆菌发酵乳中的活菌数、降低发酵乳的pH，并提高其持水力；响应面分析发现低聚半乳糖发酵乳的最佳制备条件为：2.0%的植物乳杆菌接种量、1.0%的低聚半乳糖添加量、发酵时间为24 h及发酵温度为42 ℃；添加低聚半乳糖的发酵乳能有效控制产肠毒素蜡样芽孢杆菌浓度在106 CFU/mL以下。该研究结果为低聚半乳糖及植物乳杆菌ZDY2013在发酵乳中的应用奠定了理论基础。
Abstract: In order to test the effect of galactooligosaccharide (GOS) on the characterization and antibacterial activity of milk fermented by Lactobacillus plantarum, the main factors affecting the fermented milk were investigated and the most suitable fermentation conditions for improving the characterization of fermented milk were obtained with response surface analysis. The enterotoxigenic Bacillus cereus HN001 was used as an indicator to study the effect of GOS addition on the antibacterial activity of L. plantarum ZDY2013 fermented milk. The results showed that L. plantarum could use GOS for metabolism in vitro and inhibit the growth of B. cereus effectively. The proper addition of GOS in milk could increase the number of viable bacteria, reduce the pH and enhance the water holding capacity of L. plantarum fermented milk. The result from response surface analysis was found that the best fermentation conditions for fermented milk were 2.0% inoculums of L. plantarum, 1.0% GOS, fermented at 42 ℃ for 24 h. Additionally, GOS addition in fermented milk could effectively control the concentration of enterotoxigenic B. cereus HN001 to below 106 CFU/mL. The results of this study provide the foundation for the application of GOS and L. plantarum ZDY2013 in fermented milk.
图 1 低聚半乳糖对植物乳杆菌ZDY2013生长代谢的影响
注：*表示P<0.05。
Figure 1. Effect of GOS on the metabolism of L. plantarum ZDY2013
图 2 植物乳杆菌利用改良培养基发酵的抑菌能力
Figure 2. The antibacterial ability of L. plantarum fermented with modified media
图 3 不同菌液接种量对植物乳杆菌发酵乳特性的影响
注：#不同接种量间的比较，*相同接种量间的比较；#，*表示P<0.05，##，**表示P<0.01，###，***表示P<0.001；####，****表示P<0.0001；图4~图6同。
Figure 3. Effect of different inoculation amount on the characterization of L. plantarum fermented milk
图 4 不同浓度低聚半乳糖对植物乳杆菌发酵乳特性的影响
Figure 4. Effect of different concentration of GOS on the characterization of L. plantarum fermented milk
图 5 发酵时间对植物乳杆菌发酵乳特性的影响
Figure 5. Effect of fermentation time on the characterization of L. plantarum fermented milk
图 6 发酵温度对植物乳杆菌发酵乳特性的影响
Figure 6. Effect of fermentation temperature on the characterization of L. plantarum fermented milk
图 7 不同因素对发酵乳活菌数的交互影响
Figure 7. The interactive influence of different factors on viable bacteria counts of fermented milk
图 8 低聚半乳糖发酵乳对蜡样芽孢杆菌的抑制作用
注：****表示P<0.0001。
Figure 8. Inhibition of GOS fermented milk on B. cereus
表 1 Box-Behnken试验因素与水平
Table 1 Factors and levels of Box-Behnken test
因素水平-101 X1接种量（%）123X2糖浓度（%）123X3发酵时间（h）122436X4发酵温度（℃）323742表 2 Box-Behnken试验设计与结果
Table 2 Design and results of Box-Behnken test
实验号X1X2X3X4活菌数（lg CFU/mL） 11124378.6523124378.7931324378.9343324378.8252212328.5162236328.5872212427.9582236428.0491224328.70103224328.74111224428.18123224428.21132112378.39142312378.63152136378.51162336378.57171212377.97183212378.34191236378.78203236378.99212124328.69222324328.77232124428.17242324428.22252224378.67262224378.78272224378.97282224378.67292224378.63表 3 回归方程的方差分析
Table 3 Variance analysis of regression equation
方差来源偏差平方和自由度均方F值P值显著性 模型2.02140.144.560.0038**X10.03910.0391.220.2886X20.04610.0461.440.2499X30.2410.247.430.0164*X40.8610.8627.280.0001***X1X20.01610.0160.490.4940X1X30.006410.00640.200.6599X1X40.00002510.0000250.00078930.9780X2X30.008110.00810.260.6209X2X40.00022510.0002250.0071040.9340X3X40.000110.00010.0031570.9560X120.000659510.00065950.0210.8873X220.00147610.0014760.0470.8322X320.3110.319.910.0071**X420.5210.5216.340.0012**残差0.44140.0320.0080失拟项0.37100.371.920.2764纯误差0.07640.019总和2.4728 注：*表示P<0.05；**表示P<0.01；***表示P<0.001。 [1]MARKOWIAK P, SLIZEWSKA K. Effects of probiotics, prebiotics, and synbiotics on human health[J]. Nutrients,2017,9(9):1−30.
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