Distribution and metabolism of ornithine in Rhodopseudomonas spheroides

1968 ◽  
Vol 170 (1020) ◽  
pp. 265-278 ◽  
Keyword(s):  
Time Course ◽  
Cytoplasmic Membrane ◽  
Bacterial Species ◽  
Subcellular Fractions ◽  
Diaminopimelic Acid ◽  
Cell Protein ◽  
Taxonomic Importance ◽  
Rhodopseudomonas Spheroides ◽  
Turn Over ◽  
Micro Organisms

Ornithine lipid was found in chromatophores, in poorly pigmented subcellular fractions from pigmented micro-organisms and in fragments from cells grown under oxygen which have no bacteriochlorophyll. Its quantitative distribution among these different subcellular fractions did not correlate with the distribution of diaminopimelic acid. It is concluded that ornithine lipid is a specific constituent of the cytoplasmic membrane as opposed to the cell wall. Calculations indicate that about 20% of the ornithine lipid in pigmented cells is not associated with chromatophores. The cytoplasmic membrane content of unpigmented cells, calculated on the basis of ornithine lipid as a marker, was 15 to 22% of the total cell protein. Radioactivity from DL-[5- 14 C] ornithine in trace amounts was rapidly incorporated into growing cells. Most of the counts were in proline, arginine and glutamic acid residues of the proteins. However, nearly all the radioactivity incorporated into lipid was still present as ornithine. [5- 14 C]Ornithine incorporated into lipid of oxygen-grown cells did not turn over when the organisms were allowed to adapt to photosynthetic conditions but the lipid from the chromatophores was radioactive. During this adaptation the content of ornithine lipid per cell doubled with respect to the phospholipid, which increased twofold. The time course of these changes was parallel to that of bacteriochlorophyll synthesis. The significance of all these results in relation to the nature and biogenesis of the chromato­phores is discussed. It is pointed out also that studies on the distribution of ornithine lipid in other bacterial species may be of taxonomic importance.

The isolation and characterization of subcellular fractions from pigmented and unpigmented cells of Rhodopseudomonas spheroides

1968 ◽  
Vol 170 (1020) ◽  
pp. 229-246 ◽  
Keyword(s):  
Nucleic Acid ◽  
Cytoplasmic Membrane ◽  
Sucrose Density Gradient ◽  
Subcellular Fractions ◽  
Diaminopimelic Acid ◽  
Pure Oxygen ◽  
Simple Method ◽  
Isolation And Characterization ◽  
Rhodopseudomonas Spheroides

Current ideas on the nature and development of the chromatophores of photosynthetic bacteria are reviewed. A simple method of obtaining purified chromatophores by sucrose density gradient centrifuging of cell-free extracts of Rhodopseudomonas spheroides is described. Such prepara­tions consist of about 60% protein, 20% phospholipid and 10% pigment, most of which is bacteriochlorophyll. Small quantities of carbohydrate, but only traces of nucleic acid, are found. The material was fairly homogeneous on electron microscopy. Rps. spheroides was also grown under pure oxygen in the dark. A particulate preparation from cells cultured under these conditions was similar to the chromatophores with respect to its high content of protein and of phospholipid but had a much greater content of nucleic acid and no bacteriochlorophyll. In addition, it contained amino sugars and diaminopimelic acid which are not found in chromatophores. These differences in chemical composition were correlated with the electron microscope appearances of the different subcellular fractions. It is concluded that the particulate preparation from cells grown under oxygen represents the cytoplasmic membrane of the micro-organism, but as isolated it is heavily contaminated with fragments of the cell wall from which it cannot be readily separated.

The separation and identification of the lipids of Rhodopseudomonas spheroides

1968 ◽  
Vol 170 (1020) ◽  
pp. 279-297 ◽  
Keyword(s):  
Phosphatidic Acid ◽  
Dry Weight ◽  
Detailed Examination ◽  
Subcellular Fractions ◽  
Anaerobic Conditions ◽  
Whole Cells ◽  
Rhodopseudomonas Spheroides ◽  
Micro Organisms

Detailed examination of the lipids of Rhodopseudomonas spheroides was undertaken since it was thought this might provide information on the biogenesis of the chromatophores. The phospholipids of purified chromatophores consist of phosphatidylethanolamine (35%), phosphatidylglycerol (34%) and phosphatidylcholine (23%); minor quantities of phosphatidic acid and of cardiolipin were also found. In addition, a sulpholipid was detected and also a new lipid containing ornithine. The relative proportions of the different phospholipids in chromatophores, in other subcellular fractions from pigmented micro-organisms and in fragments from cells grown under oxygen were similar. However, the ‘aerobic fragments’ contained much less ornithine lipid than the chromatophores. Comparison of the relative amounts of the different phospholipids in whole cells grown under oxygen, under air, or under semi-anaerobic conditions in the light showed no marked differences, the composition being similar to that found in chromatophores. It is concluded that there is no particular lipid specifically associated with chromatophores. Poly- β -hydroxybutyrate accounted for as much as 35% of the dry weight of cells grown under oxygen in the dark on malate-glutamate medium. It is necessary to remove this material before chromatography as it otherwise interferes with the separation.

Molecular identification of gut lactic acid bacteria in pigs by macro-arraying techniques

2005 ◽  
Vol 2005 ◽  
pp. 94-94
Author(s):  
N. Thanantong ◽  
W. Wattanakul ◽  
K. Hillman ◽  
S. Edwards ◽  
O. Sparagano
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bacterial Species ◽  
Genetic Material ◽  
Phenotypic Properties ◽  
Negative Results ◽  
Cytoplasmic Proteins ◽  
Protein Fingerprinting ◽  
Micro Organisms

Lactic acid bacteria (LAB) consist of many genera, which contain numerous bacterial species. The LAB are Gram-positive, non-spore forming micro-organisms and typically give negative results to the catalase test (Stiles and Holzapfel, 1997). The current classification of LAB combines both phenotypic properties and genotypic examination. Phenotypic studies use the cell wall compositions (mainly for Bifidobacteria), protein fingerprinting which analyse the total soluble cytoplasmic proteins, and the patterns of certain isoenzymes. The gold-standard molecular method to identify LAB is DNA-DNA homology analysis, and molecular methods using specific genetic material patterns of LAB are increasingly being applied as an identification tool. The objective of this study was to develop potential specific oligonucleotide probes for the macro-array identification of LAB.

Metabolic turnover of the lipids of Rhodopseudomonas spheroides

1968 ◽  
Vol 170 (1020) ◽  
pp. 311-318 ◽  
Keyword(s):  
Cytoplasmic Membrane ◽  
Specific Radioactivity ◽  
Membrane System ◽  
Anaerobic Conditions ◽  
Metabolic Turnover ◽  
Rhodopseudomonas Spheroides ◽  
Different Parts

Experiments with L-[Me- 14 C]methionine and inorganic 32 P indicate that the lipids of this micro-organism are metabolically stable. Lipids of unpigmented cells were incorporated into chromatophores when these cells were allowed to adapt to semi-anaerobic conditions in the light. The specific radioactivity of the phospholipid of the chromatophores was the same as that of the adapted cells; thus it appears that no distinction can be made between different parts of the cytoplasmic membrane system with respect to the labelling of the lipids. It is concluded that chromatophores originate from the cytoplasmic membrane and remain structurally a part of it.

Transcription Factors Regulating T-Cells: Implications for Graft vs. Host Disease.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3929-3929
Author(s):  
R. P. Weitzel ◽  
Y. Huang ◽  
L. Fanning ◽  
M. Kozik ◽  
P. Leahy ◽  
...  
Keyword(s):  
T Cells ◽  
Transcription Factors ◽  
T Cell ◽  
Lower Incidence ◽  
Time Course ◽  
Critical Role ◽  
Cell Protein ◽  
Rt Pcr ◽  
Host Disease ◽  
Cyclophilin B

Abstract Background. Graft versus host disease (GVHD) after allogeneic stem cell transplantation is associated with significant morbidity and mortality and targeted therapies are needed. The transcription factor nuclear factor of activated T-cells 1 (NFAT1) is known to interact with various other proteins, including AP-1 (fos/jun heterodimer), regulating both active immune responses and T-cell anergy. We have previously demonstrated lower expression of NFAT-associated transcripts via microarray, including IFN-γ, TNF-α, M-CSF, IL-3, 4, 5, and 13, IL-2R-α, CD40L, MIP1-α, as well as related transcription factors such as JunB, FOSL1, STAT4, T-bet, and c-maf in umbilical cord blood (UCB) CD4+ cells following primary stimulation when compared to cells obtained from adult blood (AB). We hypothesize a critical role for these NFAT1-dependent factors in the increased proliferation, decreased cytokine production seen in UCB, and lower incidence of GVHD following UCB transplantation. Here we present data more closely exploring the behavior of NFAT1 and associated genes in both UCB and AB CD4+ T-cells following primary stimulation. Methods. siRNA targeting NFAT1 mRNA was utilized to study the effects of lowered NFAT1 in primary human T-cells. CD14neg/4+ T-cells were isolated from adult peripheral blood by ficoll density gradient and selection by MACS. These cells were then immediately transfected via Nucleofector electroporation (Amaxa Biosystems, Gaithersburg, MD) with siRNA duplexes targeting NFAT1 and Cyclophilin B as well as an eGFP-encoding plasmid along with appropriate controls. Cultures were stimulated by anti-CD3/CD28 for 16h. 24h following transfection. Whole cell protein was harvested from a portion of each culture, and the remaining cells washed with PBS and replaced in non-stimulating media. Protein was extracted from the remaining cells 24h later. Protein lysate was analyzed by western blot for NFAT1, Cyclophilin-B, and β-Actin. Separately, expression of 7 transcripts (FOS, JUN, JUNB, FOSL1, BACH2, NFAM, and NFAT1) was analyzed via real-time PCR (RT-PCR). mRNA was obtained from 4 UCB and 4 AB samples at baseline (0h) (n=2) and 16h (n=2) of stimulation, using β-2-microglobulin as an endogenous control. Results. Transfection efficiency was measured to be approximately 70% via fluorescence microscopy of the eGFP-transfected culture. Our data indicate marginal reduction of NFAT1 protein at 24h. Notably after 48h approximately 65% knockdown is evident. Time course studies to determine the stability of siRNA-mediated NFAT1 knockdown are ongoing. No significant increase in the level of NFAT1 mRNA as measured by RT-PCR was detected. This suggests that the increase in NFAT1 protein levels post stimulation may not be a result of transcriptional regulation. Both UCB and AB samples exhibited a strong (15 fold) increase in FOSL1 transcription, and confirmed the enhanced up-regulation (about 3 fold) in the AB sample seen in microarray. Conclusions. Our data elucidate differences in the transcriptional program between UCB and AB T-cells, suggesting a crucial role for post-transcriptional events in the regulation of NFAT1 and its associated genes. Our findings identify differing regulation and expression of transcription factors in UCB vs. AB T-cells that may underlie the lower incidence and severity of GVHD seen in patients infused with UCB-derived grafts, and implicate additional elements which may modulate UCB T-cell alloreactivity. Studies to analyze the direct effects of decreased NFAT1 expression on relevant cytokines and other signaling molecules are currently ongoing.

Adaptation of Rhodopseudomonas spheroids

1968 ◽  
Vol 171 (1022) ◽  
pp. 111-125 ◽  
Keyword(s):  
Soluble Protein ◽  
Cytoplasmic Membrane ◽  
Photosynthetic Apparatus ◽  
Anaerobic Conditions ◽  
Reserve Material ◽  
Major Increase ◽  
Rhodopseudomonas Spheroides ◽  
In Cells

The composition of Rhodopseudomonas spheroides grown under oxygen or under air in the dark, or semi-anaerobically in the light was studied. The amounts of various constituents were expressed per cell. Anaerobically grown cells were 20 % lighter than cells grown under oxygen but contained approximately 50 % more protein and 85 % more phospholipid. The differences in weight were largely due to greater amounts of reserve material (poly-B-hydroxybutyrate and carbohydrate) in oxygen-grown cells. The major increase in the protein was due to a doubling of the 4particulate5 protein, the ‘soluble’ protein increasing by only 20% . The values of certain constituents in cells grown in air were intermediate between those of oxygen-grown cells and semi-anaerobically grown cells. The changes in composition were followed during adaptation from growth under oxygen to semi-anaerobic conditions in the light. Particulate protein, phospholipid, and enzymes concerned in bacteriochlorophyll synthesis increased markedly before photopigments or chromatophores were formed. These results indicate the sequence of some of the steps concerned in the differentiation of the cytoplasmic membrane of pigment-free microorganisms into the fully formed photosynthetic apparatus.

Vagococcus entomophilus sp. nov., from the digestive tract of a wasp (Vespula vulgaris)

2014 ◽  
Vol 64 (Pt_3) ◽  
pp. 731-737 ◽  
Author(s):  
J. Killer ◽  
P. Švec ◽  
I. Sedláček ◽  
J. Černohlávková ◽  
O. Benada ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Bacterial Species ◽  
Cellular Fatty Acid ◽  
Protein Profiling ◽  
Cell Protein ◽  
Phenotypic Characterization ◽  
Rrna Gene ◽  
Gene Sequences ◽  
Content Type ◽  
Vespula Vulgaris

Three unknown Gram-stain-positive, catalase-negative, facultatively anaerobic and coccus-shaped strains of bacteria were isolated from the digestive tracts of wasps (Vespula vulgaris). Analysis of 16S rRNA gene sequences revealed that these strains had identical sequences and showed that Vagococcus salmoninarum , with 96.2 % sequence similarity, was the closest phylogenetic neighbour. Further analyses based on hsp60 and pheS gene sequences of representatives of the family Enteroccocaceae and genotypic and phenotypic characterization using (GTG)5-PCR fingerprintings, EcoRI ribotyping, DNA G+C content, whole-cell protein profiling, cellular fatty acid profiles analysis and extensive biotyping confirmed that the investigated strains were representatives of a novel bacterial species within the genus Vagoccocus for which the name Vagoccocus entomophilus sp. nov. is proposed. The type strain is VOSTP2T ( = DSM 24756T = CCM 7946T).

scholarly journals THE IDENTIFICATION OF BACTERIAL SYMBIONT’S OF THE LARVAE ORYCTES RHINOCEROS L. AND THE ROLE OF THE BACTERIA IN COMPOSTING PROCESS

2018 ◽  
Vol 1 (2) ◽  
pp. 43 ◽  
Author(s):  
Octanina Sari Sijabat ◽  
Marheni Marheni ◽  
Darma Bakti
Keyword(s):  
16S Rrna ◽  
16S Rdna ◽  
Plant Material ◽  
Bacterial Species ◽  
Symbiotic Bacteria ◽  
Bacterial Symbionts ◽  
Speed Up ◽  
Hind Gut ◽  
Micro Organisms

AbstractOryctes rhinoceros L. has symbioses with micro organisms in their hind guts which further break down plant material consumed by beetle. The aim of this research is to determine the identification of the existence of the bacterial species in the hind gut larvae of the symbiotic bacteria using biochemical test and analysis based on 16S rRNA. The result of this research indicate that there were two different bacterials: Bacillus siamensis and Bacillus stratosphericus found. The bacteria was used for starting the composting and more specifically, the Bacillus siamensis can speed up composting with the end result at C/N 13.16.Keywords: Larvae O. rhinoceros L, Bacterial Symbionts, 16S rDNA, Composting

scholarly journals Bio-remedial Effect of Specific Micro-organisms (Bacteria) on Used Engine Oil: A Case Study of Some Mechanic Workshops in Maiduguri Metropolitan Council (MMC)

2020 ◽  
Vol 1 (3) ◽  
Author(s):  
Ibrahim Yerima ◽  
G. A. Felix ◽  
M. I. Ahmad
Keyword(s):  
Zea Mays ◽  
Agar Medium ◽  
Bacterial Species ◽  
Soil Samples ◽  
Engine Oil ◽  
Bacillus Sp ◽  
Used Engine Oil ◽  
Micro Organisms ◽  
Effective Growth

The potential of three micro-organisms (Pseudomonas, Streptococcus and Bacillus sp) were isolated from hydrocarbon contaminated soil and were evaluated for their biodegradation ability. The rate of biodegradation of the engine oil in the soil samples were exposed to used engine oil with different exposure rates of 5,10,15 and 20 years  were studied for a period of three (3) weeks under greenhouse experiment. The soil samples were obtained from four different mechanic workshops in M.M.C and they were plated on nutrients agar and oil agar medium to isolate the bacterial species from the spilled soil samples. All the micro-organisms used in this study showed their abilities to remediate soil exposed to used engine oil and the remediated soil samples were able to support the growth of Maize ( Zea mays) after 10 years  effective growth

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