Scott C C, Makula S R, Finnerty W R
J Bacteriol. 1976 Jul;127(1):469-80. doi: 10.1128/jb.127.1.469-480.1976.
Membranes were isolated and purified from nutrient broth-yeast extract- and hexadecane-grown cells of Acinetobacter sp. strain HO1-N. Two membrane fractions were isolated from nutrient broth-yeast extract-grown cells, the cytoplasmic membrane and the outer membrane. In addition to these two membrane fractions, a unique membrane fraction was isolated from hexadecane-grown cells (band 1) and characterized as a lipid-rich, low-density membrane containing high concentrations of hexadecane. The outer membrane preparations of Acinetobacter, obtained from nutrient broth-yeast extract- and hexadecane-grown cells, exhibited a low ratio of lipid phosphorus to protein and contained phospholipase activity and 2-keto-3-deoxyoctulosonic acid. Phosphatidic acid cytidyltransferase, adenosine triphosphatase, and reduced nicotinamide adenine dinucleotide oxidase were recovered almost exclusively in the cytoplasmic membrane fractions. The cytoplasmic membrane fractions contained 20 to 25 polypeptide species on sodium dodecyl sulfate-polyacrylamide gels, and the outer membrane fractions contained 15 to 20 polypeptide species. A major polypeptide species with an apparent molecular weight of approximately 42,000 to 44,000 was found for all outer membrane fractions. The buoyant densities of the cytoplasmic membrane fractions and the outer membrane fractions were closely similar, necessitating their separation by differential centrifugation. Band 1 of hexadecane-grown cells had a ratio of lipid phosphorus to protein that was almost twice that of cytoplasmic membrane and a correspondingly low buoyant density (1.086 g/cm3). Enzyme activities associated with band 1 were identical to those associated with the cytoplasmic membrane. The electrophoretic banding pattern of band 1 was essentially identical to the banding pattern of the cytoplasmic membrane. The phospholipid and neutral lipid compositions of the isolated membrane fractions were determined as qualitatively similar, with significant quantitative differences. The ultrastructure characteristics of the respective membrane fractions were examined by the negative-stain technique.
从营养肉汤 - 酵母提取物培养以及十六烷培养的不动杆菌属HO1 - N菌株细胞中分离并纯化细胞膜。从营养肉汤 - 酵母提取物培养的细胞中分离出两个膜组分，即细胞质膜和外膜。除了这两个膜组分外，还从十六烷培养的细胞中分离出一个独特的膜组分（条带1），其特征为富含脂质、低密度的膜，含有高浓度的十六烷。从营养肉汤 - 酵母提取物培养以及十六烷培养的细胞中获得的不动杆菌外膜制剂，脂质磷与蛋白质的比例较低，含有磷脂酶活性和2 - 酮 - 3 - 脱氧辛酮酸。磷脂酸胞苷转移酶、三磷酸腺苷酶和还原型烟酰胺腺嘌呤二核苷酸氧化酶几乎只在细胞质膜组分中回收。在十二烷基硫酸钠 - 聚丙烯酰胺凝胶上，细胞质膜组分含有20至25种多肽，外膜组分含有15至20种多肽。所有外膜组分都发现一种表观分子量约为42,000至44,000的主要多肽。细胞质膜组分和外膜组分的浮力密度非常相似，需要通过差速离心进行分离。十六烷培养细胞的条带1脂质磷与蛋白质的比例几乎是细胞质膜的两倍，相应地浮力密度较低（1.086 g/cm³）。与条带1相关的酶活性与细胞质膜相关的酶活性相同。条带1的电泳条带模式与细胞质膜的条带模式基本相同。对分离的膜组分的磷脂和中性脂质组成进行定性测定，发现它们相似，但存在显著的定量差异。通过负染技术检查了各个膜组分的超微结构特征。
