Lee K Y, So J S, Heo T R
Department of Biological Engineering, Inha University, Yonghyun-dong 253, Nam-gu, Inchon 402-751, South Korea.
J Microbiol Methods. 2001 May;45(1):1-6. doi: 10.1016/s0167-7012(01)00214-7.
This study presents a simple and fast method for the identification of bifidobacteria using a thin layer chromatographic (TLC) analysis of the short chain fatty acids in a culture broth. When the chromatogram was sprayed with the indicator solution (methyl red-bromophenol blue in 70% ethanol), lactic acid exhibited two red spots, and acetic acid, propionic acid, and butyric acid all produced blue spots. Succinic acid and citric acid produced yellow and dark yellow spot, respectively. In addition, these organic acids showed different R(f) values. The total time taken to analyze the organic acids in the 10 bacterial culture broths using the proposed method was approximately 50 min. The proposed TLC method was used to analyze the organic acids in culture broths of the following strains, five Bifidobacterium species. (Bifidobacterium longum, B. breve, B. infantis, B. bifidum, and B. adolescentis) and five other lactic acid bacteria strains (Lactobacillus casei, L. bulgaricus, L. acidophilus, Streptococcus thermophilus, and S. lactis). Both spots of lactic acid and acetic acid were detected on all the TLC plates from the five bifidobacterial culture broths. The five other lactic acid bacterial culture broths, however, only exhibited lactic acid spots. Accordingly, the proposed TLC method would appear to be a useful tool for rapid identification of Bifidobacterium spp. at the genus level.
本研究提出了一种简单快速的双歧杆菌鉴定方法，即通过对培养液中的短链脂肪酸进行薄层色谱（TLC）分析。当用指示剂溶液（70%乙醇中的甲基红-溴酚蓝）喷洒色谱图时，乳酸呈现两个红色斑点，而乙酸、丙酸和丁酸均产生蓝色斑点。琥珀酸和柠檬酸分别产生黄色和深黄色斑点。此外，这些有机酸显示出不同的比移值（R(f)值）。使用该方法分析10种细菌培养液中的有机酸，总用时约50分钟。所提出的TLC方法用于分析以下菌株培养液中的有机酸，5种双歧杆菌（长双歧杆菌、短双歧杆菌、婴儿双歧杆菌、两歧双歧杆菌和青春双歧杆菌）以及5种其他乳酸菌菌株（干酪乳杆菌、保加利亚乳杆菌、嗜酸乳杆菌、嗜热链球菌和乳酸链球菌）。在来自5种双歧杆菌培养液的所有TLC板上均检测到乳酸和乙酸的斑点。然而，其他5种乳酸菌培养液仅显示出乳酸斑点。因此，所提出的TLC方法似乎是在属水平快速鉴定双歧杆菌属的有用工具。
