Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.
PLoS One. 2013 May 20;8(5):e63816. doi: 10.1371/journal.pone.0063816. Print 2013.
In Taiwanese alternative medicine Lu-doh-huang (also called Pracparatum mungo), mung beans are mixed with various herbal medicines and undergo a 4-stage process of anaerobic fermentation. Here we used high-throughput sequencing of the 16S rRNA gene to profile the bacterial community structure of Lu-doh-huang samples. Pyrosequencing of samples obtained at 7 points during fermentation revealed 9 phyla, 264 genera, and 586 species of bacteria. While mung beans were inside bamboo sections (stages 1 and 2 of the fermentation process), family Lactobacillaceae and genus Lactobacillus emerged in highest abundance; Lactobacillus plantarum was broadly distributed among these samples. During stage 3, the bacterial distribution shifted to family Porphyromonadaceae, and Butyricimonas virosa became the predominant microbial component. Thereafter, bacterial counts decreased dramatically, and organisms were too few to be detected during stage 4. In addition, the microbial compositions of the liquids used for soaking bamboo sections were dramatically different: Exiguobacterium mexicanum predominated in the fermented soybean solution whereas B. virosa was predominant in running spring water. Furthermore, our results from pyrosequencing paralleled those we obtained by using the traditional culture method, which targets lactic acid bacteria. In conclusion, the microbial communities during Lu-doh-huang fermentation were markedly diverse, and pyrosequencing revealed a complete picture of the microbial consortium.
在台湾的替代医学中，绿豆（也称为朴硝）与各种草药混合，并经历一个四阶段的厌氧发酵过程。在这里，我们使用 16S rRNA 基因高通量测序来描绘卢豆黄样品的细菌群落结构。对发酵过程中 7 个时间点获得的样品进行焦磷酸测序，揭示了 9 个门、264 个属和 586 个种的细菌。当绿豆在竹节内（发酵过程的第 1 和第 2 阶段）时，乳杆菌科和乳杆菌属的数量最多；植物乳杆菌广泛分布于这些样品中。在第 3 阶段，细菌分布转向卟啉单胞菌科，而丁酸单胞菌成为主要的微生物成分。此后，细菌数量急剧下降，第 4 阶段的细菌数量太少而无法检测到。此外，用于浸泡竹节的液体的微生物组成差异很大：发酵大豆溶液中以极端小球菌为主，而流动泉水中则以丁酸单胞菌为主。此外，我们的焦磷酸测序结果与我们通过传统培养方法（针对乳酸菌）获得的结果一致。总之，卢豆黄发酵过程中的微生物群落非常多样，焦磷酸测序揭示了微生物联合体的全貌。
