Zhao Nan, Cai Jialiang, Zhang Chuchu, Guo Zhuang, Lu Wenwei, Yang Bo, Tian Feng-Wei, Liu Xiao-Ming, Zhang Hao, Chen Wei
a State Key Laboratory of Food Science and Technology, School of Food Science and Technology , Jiangnan University , Wuxi , P.R. China.
d Northwest Hubei Research Institute of Traditional Fermented Food, College of Chemical Engineering and Food Science, Hu Bei University of Arts and Science , Xiangyang , China.
Bioengineered. 2017 Sep 3;8(5):642-650. doi: 10.1080/21655979.2017.1300736. Epub 2017 Apr 14.
Traditional paocai brine (PB), which is continuously propagated by back-slopping and contains various species of lactic acid bacteria (LAB), is critical for the flavor of paocai. Culture-independent approaches are commonly used to investigate the microbial communities of fermented food. To evaluate the influence of different DNA (DNA) extraction methods on estimates of bacterial community profiles from 4 PBs, the lysis efficiency, DNA yield, purity and denaturing gradient gel electrophoresis (DGGE) profiles of V3 region of a 16S ribosomal ribonucleic acid gene were acquired. The cell lysis pattern of SDS + beads and Lysing matrix E+ beads (methods 3 and 4) showed higher cell lysis efficiency than SDS and SDS + Lysozyme (methods 1 and 2) in all PBs. SDS + beads obtained the largest DNA yield of the 4 methods. Moreover, methods 3 and 4 resulted in higher H' values and generated more global bacteria profiles than other methods. Overall, our results demonstrate that the properties of PB significantly affect the efficiency of DNA extraction methods. Methods 3 and 4 were both suitable for DNA extraction from PB. Method 3 is more economic, simple and rapid than method 4 for large-scale studies of the bacterial profiles of PB.
传统泡菜卤水（PB）通过回接连续传代培养，含有多种乳酸菌（LAB），对泡菜风味至关重要。不依赖培养的方法常用于研究发酵食品的微生物群落。为评估不同DNA提取方法对4种PB细菌群落图谱估计的影响，获取了16S核糖体核糖核酸基因V3区域的裂解效率、DNA产量、纯度及变性梯度凝胶电泳（DGGE）图谱。在所有PB中，SDS + 珠磨法和裂解基质E + 珠磨法（方法3和4）的细胞裂解模式显示出比SDS法和SDS + 溶菌酶法（方法1和2）更高的细胞裂解效率。SDS + 珠磨法在4种方法中获得的DNA产量最高。此外，方法3和4比其他方法产生更高的H'值，并生成更多的整体细菌图谱。总体而言，我们的结果表明PB的性质显著影响DNA提取方法的效率。方法3和4都适用于从PB中提取DNA。对于PB细菌图谱的大规模研究，方法3比方法4更经济、简单和快速。
