The “Free” Lipids of Two Different Strains of Hydrogen‐Oxidizing Bacteria in Relation to Their Growth Phases

Thiele, O. W.; Dreysel, Jörg; Hermann, Dieter

doi:10.1111/j.1432-1033.1972.tb01979.x

Cited by 27 publications

(15 citation statements)

References 56 publications

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“…The size of the layer (3 to 20 nm), however, appears to be species dependent. These studies, along with biochemical analysis of changes in the amounts of lipid during PHB production (19), suggest that a monolayer of lipid might be involved in surface coverage of granules. Recent atomic force microscopy analyses suggested that there is a structure on the granule surface that is 350 Å in diameter and has a 150-Å pore-like structure in the center (3).…”

Section: Discussionmentioning

confidence: 99%

Analysis of Transient Polyhydroxybutyrate Production in Wautersia eutropha H16 by Quantitative Western Analysis and Transmission Electron Microscopy

Tian¹,

He²,

Lawrence

et al. 2005

J Bacteriol

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Polyhydroxybutyrates (PHBs) are polyoxoesters generated from (R)3-hydroxybutyryl coenzyme A by PHB synthase. During the polymerization reaction, the polymers undergo a phase transition and generate granules. Wautersia eutropha can transiently accumulate PHB when it is grown in a nutrient-rich medium (up to 23% of the cell dry weight in dextrose-free tryptic soy broth [TSB]). PHB homeostasis under these growth conditions was examined by quantitative Western analysis to monitor the proteins present, their levels, and changes in their levels over a 48-h growth period. The proteins examined include PhaC (the synthase), PhaP (a phasin), PhaR (a transcription factor), and Polyhydroxybutyrates (PHBs) are biodegradable polymers with properties of thermoplastics synthesized by PHB synthases (PhaC) using (R)3-hydroxybutyryl coenzyme A as a substrate. In times of nutrient limitation, many bacteria generate these polymers when an appropriate carbon source is available (1). The soluble (R)3-hydroxybutyryl coenzyme A is transformed into insoluble polymers, called granules, in a nontemplate-dependent polymerization process (18). When the bacteria find themselves in a more hospitable environment, they degrade this polymer to generate building blocks and reducing equivalents for anabolism. However, bacteria in a nutrient-rich medium can also make and degrade PHB. The biology of PHB production and utilization under nutrient-rich conditions, however, remains to be elucidated.The widespread detection of genes in many microorganisms that carry out PHB synthesis and degradation suggests that these pathways have evolved to be a general mechanism for bacterial cell survival in times of stress. The organism that we have chosen to study PHB homeostasis is Wautersia eutropha H16. This organism contains a class I synthase to generate PHB (15).In this paper, we describe studies of PHB production and utilization by W. eutropha H16 in a nutrient-rich medium (dextrose-free tryptic soy broth [TSB]). Using antibodies (Abs) to most of the proteins identified as being involved in PHB homeostasis thus far, we carried out Western blotting as a function of growth time to measure the rates of appearance and disappearance of PhaC, PhaP (a phasin protein), and PhaR (a putative transcription factor), as well as PhaZ1 a , PhaZ1 b , PhaZ1 c , and PhaZ2 (formerly known as PhaZ1, PhaZ2, PhaZ3, and oligomer hydrolase, respectively); all of these proteins are thought to be involved in PHB degradation (10, 23). The average cell volume (20) and the average number of granules per cell at 4 and 24 h in TSB were determined by stereology analyses of transmission electron microscopy (TEM) images. These results and knowledge concerning the M w of PHB allowed us to estimate the amount of each protein per cell, the percentage of the granule surface that is covered by PhaC and PhaP, and the number of PhaC molecules per PHB polymer. A comparison of these results with similar studies performed under nitrogen-limited growth conditions (unpublished data) provided insight int...

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Section: Discussionmentioning

confidence: 99%

Analysis of Transient Polyhydroxybutyrate Production in Wautersia eutropha H16 by Quantitative Western Analysis and Transmission Electron Microscopy

Tian¹,

He²,

Lawrence

et al. 2005

J Bacteriol

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“…Major phospholipids were fractionated by two-dimensional thinlayer chromatography of the polar lipid fraction [8]. Lipid spots on chromatoplates were visualized by iodine vapor.…”

Section: Quan T Ifica T Ion Of Phospholipidsmentioning

confidence: 99%

“…Lipid spots on chromatoplates were visualized by iodine vapor. Estimation of relative amounts of the major phospholipids was performed by determination of the phosphorus content of each spot [8]. Phosphorus was assayed according to King [9].…”

Section: Quan T Ifica T Ion Of Phospholipidsmentioning

confidence: 99%

Occurrence of Phosphatidylcholine in Hydrogen‐Oxidizing Bacteria

Thiele

Oulevey

1981

European Journal of Biochemistry

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15 strains of hydrogen‐oxidizing bacteria were grown heterotrophically, harvested during the stationary phase of growth, and analyzed for their principal phospholipids. All strainsndash;with the exception of Corynebacterium autotrophicum strain SA 32 and Pseudomonas pseudoflava–contained phosphatidylcholine as a major constituent. It is concluded that the presence of phosphatidy1choline is neither characteristic of a peculiar bacterial genus or family, nor is it absolutely correlated to the ability to oxidize hydrogen. The phosphatidylcholines of all strains contain C19 cyclopropane acid which is, in some strains, predominantly located at C‐2 position of the glycerol moiety.

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“…Thin-layer chromatographic procedures were essentially the same as described in detail recently [16]. Additionally …”

Section: Chromatographic Methodsmentioning

confidence: 99%

The Free Lipids of Brucella melitensis and Bordetella pertussis

Thiele

Schwinn

1973

European Journal of Biochemistry

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Free lipids were extracted from BrzLcella melitensis and Bordetella pertussis and their compositions compared with one another. The bacteria contained 1.60/, and 8.101, free lipids by dry weight of the respective species.The major component of the neutral lipids of Bruc. melitensis was coenzyme Qlo (0.6 pmollg bacterial dry weight). Smaller amounts of 1,2-and 1,3-diglycerides and of fatty acyl monoesters of ethanediol and of unknown or-glycols have been found among the neutral lipids of Bruc. melitensis. The major component of the neutral lipids of Bord. pertussis was a fraction of unesterified fatty acids. Moreover, coenzyme Qs (0.2 pmol/g bacterial dry weight) was identified in Bord. pertussis. Almost all saponifiable lipids of Bruc. melitensis contained considerable amounts of a GI,-cyclopropane acid (lactobacillic acid) whereas no cyclopropane acid was found among the unesterified fatty acids of Bord. pertussis.From the polar lipid fraction of Bruc. melitensis diphosphatidylglycerol (cardiolipin), phosphatidylglycerol, phosphatidylethanolamine (cephalin), phosphatidylcholin (lecithin) and phosphatidylserine were isolated ; the phosphatidylethanolamine fraction also contained phosphatidyl-N-methylethanolamine and phosphatidyl-N,N-Jimethylethanolamine. Phosphatidylcholine was the major component (37.6O/, of all phospholipids) in Bruc. melitensis. Determination of positional distribution of the component fatty acids revealed lactobacillic acid to be preferentially located in the number 2-positions of all phospholipids, while the I-positions were preferentially occupied by palmitic acid and in most phospholipids, also by hexadecenoic acid.From Bord. pertussis, diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylserine were isolated, but no phosphatidylcholine, no partially methylated phosphatidylethanolamine, and no phosphatidylinositol were found. Palmitic acid was located more frequently at the number 1-positions, hexadecenoic acid more frequently at the number .%positions of the phospholipids. Diphosphatidylglycerol and phosphatidylethanolamine probably contained minor amounts of a GI,-cyclopropane acid.An ornithine-containing lipid was isolated from Bruc. melitemis and Bord. pertussis. Besides ornithine these lipids contained ethanediol and fatty acids. A similar finding is recorded in Bruc. abortus. Ethanediol appeared to be ester linked to ornithine and to a fatty acid. Another fatty acid is probably linked to one of the amino groups of ornithine by an amide bond. I n Bruc. melitensis ond Bruc. abortus lactobacillic acid is predominantly ester linked, while palmitic and stearic acids are predominantly amide linked. There is no lactobacillic acid at all in the ornithine lipid of Bord. pertussis. No hydroxy fatty acid has been found in the ornithine lipids of any strain.From the lipid patterns of Bruc. melitensis, of Bruc. abortus studied previously, and of Bord. pertussis we conclude that it is probably not justified to include Brucellae and Bordetellae in the one natural family of Brucellacea...

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The “Free” Lipids of Two Different Strains of Hydrogen‐Oxidizing Bacteria in Relation to Their Growth Phases

Cited by 27 publications

References 56 publications

Analysis of Transient Polyhydroxybutyrate Production in Wautersia eutropha H16 by Quantitative Western Analysis and Transmission Electron Microscopy

Analysis of Transient Polyhydroxybutyrate Production in Wautersia eutropha H16 by Quantitative Western Analysis and Transmission Electron Microscopy

Occurrence of Phosphatidylcholine in Hydrogen‐Oxidizing Bacteria

The Free Lipids of Brucella melitensis and Bordetella pertussis

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