Phospholipids from Bacillus stearothermophilus

Abstract

The lipids of Bacillus stearothermophilus strain 2184 were extracted with chloroform-methanol and separated into neutral lipid and three phospholipid fractions by chromatography on silicic acid columns. The phospholipids were identified by specific staining reactions on silicic acid-impregnated paper, by chromatography of alkaline and acid hydrolysis products, and by determination of acyl ester glycerol: nitrogen: phosphorus molar ratios. The total extractable lipid was 8% of the dry weight of whole cells and consisted of 30 to 40% neutral lipid and 60 to 70% phospholipid. The phospholipid consisted of diphosphatidyl glycerol (23 to 42%), phosphatidyl glycerol (22 to 39%), and phosphatidyl ethanolamine (21 to 32%). The concentrations of diphosphatidyl glycerol and phosphatidyl glycerol were lower in 2-hr cells than in 4-and 8-hr cells. Whole cells were fractionated by sonic treatment and differential centrifugation. The total lipid content, expressed in per cent of dry weight of each fraction was: whole protoplasts, 10%; membrane fraction, 18%; 30,000 x g particulate fraction, 22%; and 105,000 x g particulate fraction, 26%. The relative phospholipid concentrations in each fraction were about the same. As had been previously reported, none of the phospholipid was stable to alkaline hydrolysis.

Effect of Temperature on the Fatty Acid Composition of Thermus aquaticus

“…The total lipids of T. aquaticus account for 20% of the total dry weight in cells grown at 70 29 C, of which the phospholipids account for only 30% of the total lipids (manuscript in preparation). Card et al (6) reported that the phospholipids of B. stearothermophilus account for only 50 to 60% of the total extractable lipids. In both these thermophiles, the amount of neutral lipids is much greater than the amount found in mesophiles.…”

Lipids of Branhamella catarrhalis and Neisseria gonorrhoeae

“…Samples of the individual lipids were deacylated by the alkaline methanolysis procedure of White (35), and the water-soluble products of the hydrolysis were chromatographed on Whatman no. 1 paper in an ascending fashion using the following solvent systems: (i) methanol-formic acid-water, 80:13:7 (vol/vol/vol) (7); (ii) n-butanol-propionic acid-water, 142:71:100 (vol/vol/vol) (4); and (iii) phenol-water, 100:38 (wt/vol) (6). Visualization on thin-layer plates was accomplished by sulfuric acid, charring iodine vapor, ninhydrin, or a-naphthol reagent for glycolipids (17).…”

Isolation and Characterization of a Phosphatidylethanolamine-Deficient Mutant of Bacillus subtilis

“…Phosphatidylserine, L-serine, and monoethanolamine proved fruitless as supplements in numerous trials. bRF values for GPGPG, GPE, and GPG are those quoted by Card et al (6). BPW, n-butanol-propionic acid-water (142:71: 100, v/v/v); n, ninhydrin-positive.…”

“…Measured with a Bausch & Lomb Spectronic 20 spectrophotometer. 6 12 18 the nature of this ability to grow on TSB is curious. We speculate that TSB provides a rich source of a battery of nutrients which the mutant cannot synthesize or retain at optimal levels in a synthetic medium.…”

“…Deacylated forms were chromatographed on Whatman no. I paper by using (i) 75% phenol and (ii) n-butanol-propionic acid-water (142:71:100, v/v/v) (6). Diacyl lipids were revealed by reaction with rhodamine 6G (20), ninhydrin (20), ammonium molybdate for phosphorus (3), and periodate-Schiff (34) and periodatebenzidine (7) for vicinal hydroxyls and aniline-acidphthalate (4).…”