PREPARATION AND CHEMICAL COMPOSITION OF CELL WALLS OF MATURE INFECTIOUS DENSE FORMS OF MENINGOPNEUMONITIS ORGANISMS
PREPARATION AND CHEMICAL COMPOSITION OF CELL WALLS OF MATURE INFECTIOUS DENSE FORMS OF MENINGOPNEUMONITIS ORGANISMS
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DOI：
10.1128/jb.94.4.1178-1183.1967
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发表时间：
1967-01-01
影响因子：
3.2
通讯作者：
TAMURA, A
中科院分区：
文献类型：
--
作者：
MANIRE, GP;TAMURA, A
较大规模的脑膜肺炎微生物的生产和纯化是为了对感染性致密形式的细胞壁进行化学和免疫学研究。通过在摇床中用玻璃珠破碎纯化的微生物，通过蔗糖密度梯度离心法、酶消化法和十二烷基硫酸钠处理得到高纯度的细胞壁制剂。从完整生物中纯化的细胞壁的干重回收率约为13%。当32P标记的细胞壁制剂被分成酸溶组分、脂类制剂、核糖核酸(RNA)、脱氧核糖核酸(DNA)和残余组分时，细胞壁制剂中约80%的32P被回收到磷脂部分，相当于完整生物磷脂总量的3%左右。在纯化的细胞壁中，约7%的32P分别在RNA和DNA组分中被回收，但这仅相当于完整生物体中这些组分中32P的约0.4%。根据干重测定，计算出纯化的细胞壁制剂只含有0.6%的总核酸，这些可能不是真正的细胞壁成分。这些细胞壁含有70%到75%的蛋白质，相当于完整生物体中蛋白质的14%左右。对这些蛋白质的氨基酸分析表明，所有常见的氨基酸、氨基葡萄糖和氨基半乳糖都存在。然而，所采用的方法均未检测到胞壁酸。
Relatively large-scale production and purification of meningopneumonitis organisms was developed for chemical and immunological studies on cell walls of the infectious dense forms. By disruption of purified organisms with glass beads in a Mickle shaker, highly purified preparations of cell walls were obtained by sucrose density gradient centrifugation, enzyme digestion, and sodium dodecyl sulfate treatment. The dry-weight recovery of purified cell walls from intact organisms was about 13%. When32P-labeled preparations of cell walls were fractionated into acid-soluble, lipid, ribonucleic acid (RNA), deoxyribonucleic (DNA), and residual fractions, about 80% of the32P in cell wall preparations was recovered in the phospholipid fraction, which corresponded to about 3% of the total phospholipid in the intact organisms. About 7% of the32P in purified cell walls was recovered in the RNA and DNA fractions respectively, but this corresponds to only about 0.4% of the32P found in those fractions in intact organisms. From dry-weight determinations, it was calculated that the purified cell wall preparations contained only 0.6% total nucleic acids, and these are probably not true cell wall constituents. These cell walls contained 70 to 75% protein, corresponding to about 14% of the protein in intact organisms. Amino acid analysis of these protein showed the existence of all common amino acids, glucosamine, and galactosamine. However, no muramic acid was detected by the methods employed.