Chair of Food Science, Institute of Nutritional Science, and Interdisciplinary Research Centre for Biosystems, Land Use and Nutrition, Justus Liebig University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany.
Adisseo France S.A.S, Immeuble Anthony Parc 2, 10 Place du Général de Gaulle, 92160 Antony, France.
J Chromatogr A. 2021 Mar 15;1640:461929. doi: 10.1016/j.chroma.2021.461929. Epub 2021 Jan 22.
Imaging high-performance thin-layer chromatography (HPTLC) was explored with regard to its ability to visualize changes in the metabolite profile of bacteria. Bacillus subtilis has become a model organism in many fields. The increasing interest in these bacteria is driven by their attributed probiotic activity. However, growth behavior and metabolism of Bacillus species have a considerable influence on their activity and secondary metabolite profile. On the HPTLC plate, cultivation broths of Bacillus species (B. subtilis, B. licheniformis, B. pumilus and B. amyloliquefaciens) and some B. subtilis strains of high genetic similarity up to 99.5% were applied directly and compared with their respective liquid-liquid extracts. The latter as well as the cultivation in a minimal medium reduced the matrix load and improved the zone resolution. Cultivation parameters such as nutrient supply, cultivation temperature, cultivation time and rotational speed (oxygen level) as well as medium change were shown to have a considerable influence on the growth behavior and resulting metabolite profiles. Imaging HPTLC turned out to be an efficient and affordable tool to visualize such influences of cultivation parameters on the metabolite profiles. It converts the complexity of reaction processes occurring during cell cultivation in easy-to-understand images, which are helpful to figure out factors of influence and understand activity changes. The results highlighted that optimal cultivation conditions need to be found for the intended bacterial application, and in particular, these conditions have to be kept constant. It must be ensured that small variations in cultivation parameters of bacteria do not change the specified (probiotic) effect on the health of animals and humans. The HPTLC metabolite profiles represented the cultivation conditions of specific bacteria and were found to be a proof of the activity of distinct bacteria. In addition, HPTLC can also be used to optimize and streamline the culture media. The quality control of cultivation or fermentation processes can benefit from such a powerful tool, as a picture is worth a thousand words.
采用高性能薄层色谱（HPTLC）研究了其可视化细菌代谢物谱变化的能力。枯草芽孢杆菌已成为许多领域的模式生物。人们对这些细菌的兴趣日益增加，这是因为它们具有益生菌活性。然而，芽孢杆菌的生长行为和代谢对其活性和次生代谢物谱有相当大的影响。在 HPTLC 板上，直接应用枯草芽孢杆菌（枯草芽孢杆菌、地衣芽孢杆菌、短小芽孢杆菌和解淀粉芽孢杆菌）和一些遗传相似度高达 99.5%的枯草芽孢杆菌菌株的培养肉汤，并将其与各自的液-液提取物进行比较。后者以及在最小培养基中的培养降低了基质负载并提高了区域分辨率。培养参数，如营养供应、培养温度、培养时间和转速（氧气水平）以及培养基变化，对生长行为和产生的代谢物谱有相当大的影响。成像 HPTLC 被证明是一种有效的、经济实惠的工具，可以直观地观察培养参数对代谢物谱的影响。它将细胞培养过程中发生的复杂反应过程转化为易于理解的图像，有助于找出影响因素并理解活性变化。结果强调，需要为预期的细菌应用找到最佳的培养条件，特别是要保持这些条件不变。必须确保细菌培养参数的微小变化不会改变其对动物和人类健康的特定（益生菌）作用。HPTLC 代谢物谱代表了特定细菌的培养条件，被认为是特定细菌活性的证明。此外，HPTLC 还可用于优化和简化培养基。这种强大的工具可用于培养或发酵过程的质量控制，因为一图胜千言。
