Chavarria-Quicaño Estefany, De la Torre-González Francisco, González-Riojas Mario, Rodríguez-González Jorge, Asaff-Torres Ali
Department of Science, Research Center in Food and Development, Hermosillo, Sonora, México.
Innovak Global, Chihuahua, Chihuahua, México.
Appl Microbiol Biotechnol. 2023 Mar;107(5-6):1537-1549. doi: 10.1007/s00253-023-12391-w. Epub 2023 Jan 31.
The aim of this work was to develop a comparative study between Bacillus paralicheniformis TB197 and B. subtilis ATCC 21332 strains in terms of growth, cyclic lipopeptide production, nematicidal activity, and active lipopeptide characteristics. Crude lipopeptide extracts (CLEs) from their fermentation broths were obtained, and their nematicidal activity (NA) was estimated as the mean lethal dose (LD), employing Caenorhabditis elegans. Using a bioguided approach, CLE components were fractionated by semipreparative thin layer chromatography, and active lipopeptides were characterized by mass spectrometry. Both strains produced similar concentrations of CLEs (p ≥ 0.05) (0.99 ± 0.11 and 1.14 ± 0.15 mg/mL by TB197 and ATCC 21332, respectively). The estimated LD values of CLEs from the TB197 and ATCC 21332 strains were 3.88 and 8.15 mg/mL, respectively, showing that the NA of the TB197 strain CLE was 2.1-fold higher (p ≤ 0.05). Mass spectrometry revealed that strain TB197 synthesizes several families of lipopeptides, namely, fengycin A (C-C), fengycin B (C-C), surfactin (C-C), and lichenysin (C, C, C, and C), from which fengycins and lichenysins possess the highest NA (100 and 60% mortality in C. elegans larvae, respectively), while the ATCC 21332 strain produces mainly surfactin (C-C) (NA 63% mortality). The main differences found in this study were that the TB197 strain has a higher tolerance to inhibition by the product, and the lipopeptides they synthesize have a higher nematicidal activity due to the diversity of families compared to ATCC 21332. Likewise, it was shown that more polar lipopeptides (fengycins) are more effective at causing mortality in C. elegans larvae. KEY POINTS: • The nematicidal activity of lipopeptides from TB197 is higher than from ATCC 21332 • TB197 produces surfactin, lichenysin, and fengycin, while ATCC 21332 mainly produces surfactin • The most polar lipopeptides (fengycins) cause more mortality in C. elegans L2.
本研究旨在对类地衣芽孢杆菌TB197和枯草芽孢杆菌ATCC 21332菌株在生长、环脂肽产生、杀线虫活性及活性脂肽特性方面进行比较研究。从它们的发酵液中获得了粗脂肽提取物（CLEs），并以秀丽隐杆线虫为实验对象，将其杀线虫活性（NA）评估为平均致死剂量（LD）。采用生物导向法，通过半制备薄层色谱对CLE成分进行分离，并利用质谱对活性脂肽进行表征。两株菌产生的CLEs浓度相似（p≥0.05）（TB197和ATCC 21332分别为0.99±0.11和1.14±0.15mg/mL）。TB197和ATCC 21332菌株CLEs的估计LD值分别为3.88和8.15mg/mL，表明TB197菌株CLE的NA高2.1倍（p≤0.05）。质谱分析表明，TB197菌株合成了几个脂肽家族，即丰原素A（C-C）、丰原素B（C-C）、表面活性素（C-C）和地衣芽孢杆菌素（C、C、C和C），其中丰原素和地衣芽孢杆菌素具有最高的NA（分别导致秀丽隐杆线虫幼虫100%和60%的死亡率），而ATCC 21332菌株主要产生表面活性素（C-C）（NA为63%死亡率）。本研究发现的主要差异在于，TB197菌株对产物抑制的耐受性更高，并且与ATCC 213相比，它们合成的脂肽由于家族多样性而具有更高的杀线虫活性。同样，研究表明，极性更强的脂肽（丰原素）在导致秀丽隐杆线虫幼虫死亡方面更有效。要点：• TB197的脂肽杀线虫活性高于ATCC 21332• TB197产生表面活性素、地衣芽孢杆菌素和丰原素，而ATCC 21332主要产生表面活性素• 极性最强的脂肽（丰原素）导致秀丽隐杆线虫L2死亡率更高
