Hsueh Yi-Huang, Somers Eileen B, Lereclus Didier, Ghelardi Emilia, Wong Amy C Lee
Department of Bacteriology, Food Research Institute, University of Wisconsin-Madison, 1550 Linden Drive, Madison, WI 53706, USA.
Appl Environ Microbiol. 2007 Nov;73(22):7225-31. doi: 10.1128/AEM.00690-07. Epub 2007 Oct 5.
Bacillus cereus ATCC 14579 can respond to nutrient changes by adopting different forms of surface translocation. The B. cereus ATCC 14579 DeltaplcR mutant, but not the wild type, formed dendritic (branched) patterns on EPS [a low-nutrient medium that contains 7.0 g K(2)HPO(4), 3.0 g KH(2)PO(4), 0.1 g MgSO(4).7H(2)O, 0.1 g (NH(4))(2)SO(4), 0.01 g CaCl(2), 0.001 g FeSO(4), 0.1 g NaCl, 1.0 g glucose, and 125 mg yeast extract per liter] containing 0.7% agar. The dendritic patterns formed by sliding translocation of nonflagellated cells are enhanced under low-nutrient conditions and require sufficient production of a biosurfactant, which appears to be repressed by PlcR. The wild-type and complemented strains failed to slide on the surface of EPS agar because of the production of low levels of biosurfactant. Precoating EPS agar surfaces with surfactin (a biosurfactant produced by Bacillus subtilis) or biosurfactant purified from the DeltaplcR mutant rescued the ability of the wild-type and complemented strains to slide. When grown on a nutrient-rich medium like Luria-Bertani agar, both the wild-type and DeltaplcR mutant strains produced flagella. The wild type was hyperflagellated and elongated and exhibited swarming behavior, while the DeltaplcR mutant was multiflagellated and the cells often formed long chains but did not swarm. Thin-layer chromatography and mass spectrometry analyses suggested that the biosurfactant purified from the DeltaplcR mutant was a lipopeptide and had a mass of 1,278.1722 (m/z). This biosurfactant has hemolytic activity and inhibited the growth of several gram-positive bacteria.
蜡样芽孢杆菌ATCC 14579可通过采用不同形式的表面易位来响应营养变化。蜡样芽孢杆菌ATCC 14579的ΔplcR突变体(而非野生型)在EPS[一种低营养培养基,每升含有7.0 g K₂HPO₄、3.0 g KH₂PO₄、0.1 g MgSO₄·7H₂O、0.1 g (NH₄)₂SO₄、0.01 g CaCl₂、0.001 g FeSO₄、0.1 g NaCl、1.0 g葡萄糖和125 mg酵母提取物]含0.7%琼脂的培养基上形成树突状(分支状)图案。非鞭毛细胞通过滑动易位形成的树突状图案在低营养条件下会增强,且需要足够量生物表面活性剂的产生,而这种生物表面活性剂的产生似乎受到PlcR的抑制。野生型和互补菌株由于生物表面活性剂产量低而无法在EPS琼脂表面滑动。用表面活性素(一种由枯草芽孢杆菌产生的生物表面活性剂)或从ΔplcR突变体中纯化的生物表面活性剂预涂EPS琼脂表面,可以挽救野生型和互补菌株的滑动能力。当在富含营养的培养基如Luria-Bertani琼脂上生长时,野生型和ΔplcR突变体菌株均产生鞭毛。野生型有过多的鞭毛且细胞伸长,并表现出群体运动行为,而ΔplcR突变体有多根鞭毛,细胞常形成长链但不群体运动。薄层色谱和质谱分析表明,从ΔplcR突变体中纯化的生物表面活性剂是一种脂肽,质量为1278.1722(m/z)。这种生物表面活性剂具有溶血活性,并能抑制几种革兰氏阳性细菌的生长。