Beney L, Mille Y, Gervais P
Laboratoire de Génie des Procédés Alimentaires et Biotechnologiques, Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation, 1 Esplanade Erasme, 21000, Dijon, France.
Appl Microbiol Biotechnol. 2004 Sep;65(4):457-64. doi: 10.1007/s00253-004-1574-x. Epub 2004 Apr 17.
This study reports the effects of exposure to increasing osmotic pressure on the viability and membrane structure of Escherichia coli. Changes in membrane structure after osmotic stress were investigated by electron transmission microscopy, measurement of the anisotropy of the membrane fluorescent probe DPH (1,6-diphenyl-1,3,5-hexatriene) inserted in E. coli, and Fourier infrared spectroscopy (FTIR). The results show that, above a critical osmotic pressure of 35 MPa, the viability of the bacterium is drastically reduced (2 log decrease in survivors). Electron micrographs revealed a severe contraction of the cytoplasm and the formation of membrane vesicles at 40 MPa. Changes in DPH anisotropy showed that osmotic dehydration to 40 MPa promoted a decrease in the membrane fluidity of integral cells of E. coli. FTIR measurements showed that at 10-40 MPa a transition from lamellar liquid crystal to lamellar gel among the phospholipids extracted from E. coli occurred. Bacterial death resulting from dehydration can be attributed to the conjunction between membrane deformation, caused by the volumetric contraction, and structural changes of the membrane lipids. The influence of the latter on the formation of membrane vesicles and on membrane permeabilization at lethal osmotic pressure is discussed, since vesiculation is hypothetically responsible for cell death.
本研究报告了暴露于不断增加的渗透压对大肠杆菌活力和膜结构的影响。通过电子透射显微镜、测量插入大肠杆菌中的膜荧光探针DPH(1,6-二苯基-1,3,5-己三烯)的各向异性以及傅里叶红外光谱(FTIR),研究了渗透胁迫后膜结构的变化。结果表明,在临界渗透压35MPa以上,细菌的活力急剧下降(存活菌数减少2个对数级)。电子显微镜照片显示,在40MPa时细胞质严重收缩并形成膜泡。DPH各向异性的变化表明,渗透脱水至40MPa会导致大肠杆菌完整细胞的膜流动性降低。FTIR测量表明,在10 - 40MPa时,从大肠杆菌中提取的磷脂发生了从层状液晶到层状凝胶的转变。脱水导致的细菌死亡可归因于体积收缩引起的膜变形与膜脂结构变化之间的共同作用。讨论了后者对膜泡形成以及在致死渗透压下膜通透性的影响,因为推测膜泡化是细胞死亡的原因。