Chen He, Niu Jinfeng, Qin Tao, Ma Qi, Wang Lei, Shu Guowei
College of Life Science and Engineering, Shaanxi University of Science and Technology, China.
Enzyme Engineering Institute, Shaanxi Academy of Sciences, China.
Acta Sci Pol Technol Aliment. 2015 Jul-Sep;14(3):227-232. doi: 10.17306/J.AFS.2015.3.24.
Lactobacillus acidophilus not only improves the intestinal flora balance but also inhabits the growth of undesirable microorganisms in intestine, which is benefit to the health of humans and animals. Plackett-Burman and steepest ascent experiment are the rapid and concise ways of screening the main effective factors. This study is aimed to select the main influence factors and optimize the medium for Lactobacillus acidophilus by Plackett-Burman experiment and steepest ascent experiment.
The ideal carbon source was screened among glucose, maltose, lactose and whey powder, and the ideal nitrogen source was screened among casein hydrolysate, peptone, yeast extract powder, fish meal, carbamide, ammonium sulfate and sodium nitrate by single factor experiment. Plackett-Burman and steepest ascent experiment were applied to screen the main effective factors of Lactobacillus acidophilus among peptone, beef extract, yeast extract powder, glucose, K2HPO4, C6H14O7N2, CH3COONa, MgSO4 and Tween-80. Result. The results indicated that glucose (p = 0.01510) as negative factor and K2HPO4 (p = 0.02017) as positive effect were the significant growth factors of Lactobacillus acidophilus, CH3COONa (p = 0.09273) as positive effect was an important factor, and the optimized medium was as follows: glucose - 21 g/L, K2HPO4 - 3.5 g/L, CH3COONa - 6.5 g/L, peptone - 10 g/L, beef extract - 8 g/L, yeast extract pow.
nd. Lactobacillus acidophilus not only improves the intestinal flora balance but also inhabits the growth of undesirable microorganisms in intestine, which is benefit to the health of humans and animals. Plackett-Burman and steepest ascent experiment are the rapid and concise ways of screening the main effective factors. This study is aimed to select the main influence factors and optimize the medium for Lactobacillus acidophilus by Plackett-Burman experiment and steepest ascent experiment. Material and methods. The ideal carbon source was screened among glucose, maltose, lactose and whey powder, and the ideal nitrogen source was screened among casein hydrolysate, peptone, yeast extract powder, fish meal, carbamide, ammonium sulfate and sodium nitrate by single factor experiment. Plackett-Burman and steepest ascent experiment were applied to screen the main effective factors of Lactobacillus acidophilus among peptone, beef extract, yeast extract powder, glucose, K2HPO4, C6H14O7N2, CH3COONa, MgSO4 and Tween-80. Result. The results indicated that glucose (p = 0.01510) as negative factor and K2HPO4 (p = 0.02017) as positive effect were the significant growth factors of Lactobacillus acidophilus, CH3COONa (p = 0.09273) as positive effect was an important factor, and the optimized medium was as follows: glucose - 21 g/L, K2HPO4 - 3.5 g/L, CH3COONa - 6.5 g/L, peptone - 10 g/L, beef extract - 8 g/L, yeast extract powder - 8 g/L, C6H14O7N2 - 2 g/L, MgSO4 - 0.2 g/L and Tween-80 - 1 mL/L when the maximum viable count could achieve 2.72·109 cfu/mL.
The experimental model is reliable and the experimental results are of good stability. Variance analysis is performed to determine the adequacy and significance of the linear model. Thus, Plackett-Burman and steepest ascent experiment improve the veracity of optimization the medium for Lactobacillus acidophilus compared with the previous research.
嗜酸乳杆菌不仅能改善肠道菌群平衡，还能抑制肠道中有害微生物的生长，这对人和动物的健康有益。Plackett-Burman实验和最速上升实验是筛选主要影响因素的快速且简洁的方法。本研究旨在通过Plackett-Burman实验和最速上升实验筛选嗜酸乳杆菌的主要影响因素并优化其培养基。
通过单因素实验在葡萄糖、麦芽糖、乳糖和乳清粉中筛选理想碳源，在酪蛋白水解物、蛋白胨、酵母提取物粉、鱼粉、尿素、硫酸铵和硝酸钠中筛选理想氮源。应用Plackett-Burman实验和最速上升实验在蛋白胨、牛肉膏、酵母提取物粉、葡萄糖、K2HPO4、C6H14O7N2、CH3COONa、MgSO4和吐温-80中筛选嗜酸乳杆菌的主要影响因素。结果：结果表明，作为负因素的葡萄糖（p = 0.01510）和作为正效应的K2HPO4（p = 0.02017）是嗜酸乳杆菌的显著生长因素，作为正效应的CH3COONa（p = 0.09273）是重要因素，优化后的培养基如下：葡萄糖-21 g/L、K2HPO4-3.5 g/L、CH3COONa-6.5 g/L、蛋白胨-10 g/L、牛肉膏-8 g/L、酵母提取物粉-8 g/L、C6H14O7N2-2 g/L、MgSO4-0.2 g/L和吐温-80-1 mL/L，此时最大活菌数可达2.72·109 cfu/mL。
实验模型可靠，实验结果稳定性良好。进行方差分析以确定线性模型的充分性和显著性。因此，与先前研究相比，Plackett-Burman实验和最速上升实验提高了优化嗜酸乳杆菌培养基的准确性。
