摘要: 為探究低聚半乳糖對(duì)植物乳桿菌發(fā)酵乳特性及抗菌活性的影響，本文采用單因素法考察影響發(fā)酵乳特性的主要因素，并以響應(yīng)面法優(yōu)化發(fā)酵乳最佳發(fā)酵條件；以產(chǎn)腸毒素蠟樣芽孢桿菌HN001為指示菌，探究低聚半乳糖的添加對(duì)植物乳桿菌ZDY2013發(fā)酵乳抑菌活性的改善作用。結(jié)果表明：植物乳桿菌能有效利用低聚半乳糖進(jìn)行體外代謝，并抑制蠟樣芽孢桿菌生長(zhǎng)；牛奶中添加適量低聚半乳糖能夠增加植物乳桿菌發(fā)酵乳中的活菌數(shù)、降低發(fā)酵乳的pH，并提高其持水力；響應(yīng)面分析發(fā)現(xiàn)低聚半乳糖發(fā)酵乳的最佳制備條件為：2.0%的植物乳桿菌接種量、1.0%的低聚半乳糖添加量、發(fā)酵時(shí)間為24 h及發(fā)酵溫度為42 ℃；添加低聚半乳糖的發(fā)酵乳能有效控制產(chǎn)腸毒素蠟樣芽孢桿菌濃度在106 CFU/mL以下。該研究結(jié)果為低聚半乳糖及植物乳桿菌ZDY2013在發(fā)酵乳中的應(yīng)用奠定了理論基礎(chǔ)。

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.

圖  1   低聚半乳糖對(duì)植物乳桿菌ZDY2013生長(zhǎng)代謝的影響

注：*表示P<0.05。

Figure  1.   Effect of GOS on the metabolism of L. plantarum ZDY2013

圖  2   植物乳桿菌利用改良培養(yǎng)基發(fā)酵的抑菌能力

Figure  2.   The antibacterial ability of L. plantarum fermented with modified media

圖  3   不同菌液接種量對(duì)植物乳桿菌發(fā)酵乳特性的影響

注：#不同接種量間的比較，*相同接種量間的比較；#，*表示P<0.05，##，**表示P<0.01，###，***表示P<0.001；####，****表示P<0.0001；圖4~圖6同。

Figure  3.   Effect of different inoculation amount on the characterization of L. plantarum fermented milk

圖  4   不同濃度低聚半乳糖對(duì)植物乳桿菌發(fā)酵乳特性的影響

Figure  4.   Effect of different concentration of GOS on the characterization of L. plantarum fermented milk

圖  5   發(fā)酵時(shí)間對(duì)植物乳桿菌發(fā)酵乳特性的影響

Figure  5.   Effect of fermentation time on the characterization of L. plantarum fermented milk

圖  6   發(fā)酵溫度對(duì)植物乳桿菌發(fā)酵乳特性的影響

Figure  6.   Effect of fermentation temperature on the characterization of L. plantarum fermented milk

圖  7   不同因素對(duì)發(fā)酵乳活菌數(shù)的交互影響

Figure  7.   The interactive influence of different factors on viable bacteria counts of fermented milk

圖  8   低聚半乳糖發(fā)酵乳對(duì)蠟樣芽孢桿菌的抑制作用

注：****表示P<0.0001。

Figure  8.   Inhibition of GOS fermented milk on B. cereus

表  1   Box-Behnken試驗(yàn)因素與水平

Table  1   Factors and levels of Box-Behnken test

因素水平-101 X1接種量（%）123X2糖濃度（%）123X3發(fā)酵時(shí)間（h）122436X4發(fā)酵溫度（℃）323742

表  2   Box-Behnken試驗(yàn)設(shè)計(jì)與結(jié)果

Table  2   Design and results of Box-Behnken test

實(shí)驗(yàn)號(hào)X1X2X3X4活菌數(shù)（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   回歸方程的方差分析

Table  3   Variance analysis of regression equation

方差來(lái)源偏差平方和自由度均方F值P值顯著性 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**殘差0.44140.0320.0080失擬項(xiàng)0.37100.371.920.2764純誤差0.07640.019總和2.4728 注：*表示P<0.05；**表示P<0.01；***表示P<0.001。 [1]

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