scholarly journals Selective isolation and characterization of rare actinomycetes adopted in glacier soil of Manali ice point and its activity against Mycobacterium spp

2017 ◽  
Vol 7 (5) ◽  
pp. 1 ◽  
Author(s):  
RAJA A ◽  
P Gajalakshmi
Keyword(s):  
Active Compound ◽  
Gradient Centrifugation ◽  
Population Isolation ◽  
Isolation And Characterization ◽  
Novel Approach ◽  
Rare Actinomycetes ◽  
Nocardia Sp ◽  
Glacier Ice ◽  
Mycobacterium Spp

An exigent demand of antimicrobial agent against M. tuberculosis was lead to the isolation of novel rare actinomycetes from the unexplored cryophilic environment. Soil samples were collected from glacier ice point of Kullu Manali and processed for further studies. A novel approach was described for the isolation of rare actinomycetes from a heterogeneous population. Isolation was done by conventional and density gradient centrifugation. Sucrose gradient centrifugation showed a maximum of 24 actinomycetes isolates which belong to the genera of Streptomyces sp (12), Micromonospora sp (5) Planomonospora sp(2), Micropolyspora sp (2), Actinopolyspora sp (1) Nocardia sp (1) and Intrasporangium sp (1). Of these 24 actinomycetes, isolate Planomonospora sp (PL-2) showed potent anti-mycobacterial activity against Mycobacterium smegmatis (MTCC300) and M.tuberculosis (MTCC 6). Bioautography reveals that the Rf value of active compound was 0.75 and retains the antimicrobial activity at 75° C. Based on the C13 and H1 NMR the active compound was characterized as 2-(2-ethenylphenyl) heptane-1-ol. Phylogenetic analysis reveals active isolate was closely related to Planomonospora alba and the Genbank accession is JQ280498.

The Isolation and Characterization of Platelet Fibringgen

1979 ◽  
Author(s):  
R.E Zimmermann
Keyword(s):  
Blood Clotting ◽  
Biochemical Analysis ◽  
Gradient Centrifugation ◽  
Physiological Role ◽  
Carbohydrate Content ◽  
Freezing And Thawing ◽  
Fibrin Formation ◽  
Isolation And Characterization ◽  
Controlled Precipitation

Since platelets contribute essentially to the blood clotting mechanism the question arises as to what extent their physiological role is expressed by a specified type of fibrinogen.Platelets are harvested , washed exhaustively and homogenated by freezing and thawing. After gradient centrifugation of the homogenate fibrinogen was isolated by controlled precipitation steps. According to biochemical analysis platelet fibrinogen is of a unique polypeptid-composition, differs in its carbohydrate content and reveals a different kinetic of fibrin formation.

scholarly journals Isolation and Characterization of Live Yeast Cells from Ancient Vessels as a Tool in Bio-Archaeology

mBio ◽  
2019 ◽  
Vol 10 (2) ◽  
Author(s):  
Tzemach Aouizerat ◽  
Itai Gutman ◽  
Yitzhak Paz ◽  
Aren M. Maeir ◽  
Yuval Gadot ◽  
...  
Keyword(s):  
Ancient Dna ◽  
Residue Analysis ◽  
Fermented Food ◽  
Yeast Cells ◽  
Fermented Beverages ◽  
Isolation And Characterization ◽  
Yeast Strains ◽  
Novel Approach ◽  
Clay Matrix

ABSTRACTAncient fermented food has been studied based on recipes, residue analysis, and ancient-DNA techniques and reconstructed using modern domesticated yeast. Here, we present a novel approach based on our hypothesis that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and their descendants could be isolated and studied today. We developed a pipeline of yeast isolation from clay vessels and screened for yeast cells in beverage-related and non-beverage-related ancient vessels and sediments from several archaeological sites. We found that yeast cells could be successfully isolated specifically from clay containers of fermented beverages. The findings that genotypically the isolated yeasts are similar to those found in traditional African beverages and phenotypically they grow similar to modern beer-producing yeast strongly suggest that they are descendants of the original fermenting yeast. These results demonstrate that modern microorganisms can serve as a new tool in bio-archaeology research.IMPORTANCESo far, most of the study of ancient organisms has been based mainly on the analysis of ancient DNA. Here we show that it is possible to isolate and study microorganisms—yeast in this case—from ancient pottery vessels used for fermentation. We demonstrate that it is highly likely that these cells are descendants of the original yeast strains that participated in the fermentation process and were absorbed into the clay matrix of the pottery vessels. Moreover, we characterized the isolated yeast strains, their genomes, and the beer they produced. These results open new and exciting avenues in the study of domesticated microorganisms and contribute significantly to the fields of bio- and experimental archaeology that aim to reconstruct ancient artifacts and products.

Isolation and characterization of a mitochondrial RNA polymerase from Drosophila melanogaster

1987 ◽  
Vol 65 (2) ◽  
pp. 173-182 ◽  
Author(s):  
Michael Goldenthal ◽  
James T. Nishiura
Keyword(s):  
Drosophila Melanogaster ◽  
Rna Polymerase ◽  
Nuclear Dna ◽  
Gradient Centrifugation ◽  
Mitochondrial Rna ◽  
Mitochondrial Rna Polymerase ◽  
Isolation And Characterization ◽  
Cscl Gradient ◽  
Nuclear Rna

A DNA-dependent RNA polymerase was solubilized from sucrose gradient isolated, DNase-treated mitochrondria of Drosophila melanogaster. The isolated mitochondria were not detectably contaminated with nuclear DNA as shown by CsCl gradient centrifugation and polylysine Kieselguhr chromatography. The detergent-solubilized RNA polymerase was sensitive to rifampicin, resistant to α-amanitin, had an apparent molecular mass of about 60 kilodaltons, and displayed a tendency to aggregate, both in crude extracts or when purified. The mitochondrial RNA polymerase could be distinguished from nuclear RNA polymerases on the basis of size, salt optima, rifampicin sensitivity, and α-amanitin resistance.

scholarly journals Fractionation and characterization of oligomeric, protofibrillar and fibrillar forms of β-amyloid peptide

2000 ◽  
Vol 348 (1) ◽  
pp. 137-144 ◽  
Author(s):  
Robin V. WARD ◽  
Kevin H. JENNINGS ◽  
Robert JEPRAS ◽  
William NEVILLE ◽  
Davina E. OWEN ◽  
...  
Keyword(s):  
Cell Viability ◽  
Hippocampal Neurons ◽  
Senile Plaques ◽  
Gradient Centrifugation ◽  
Neuronal Cell ◽  
Amyloid Peptide ◽  
Aβ Peptide ◽  
Isolation And Characterization ◽  
Β Amyloid

The β-amyloid (Aβ) peptide, a major component of senile plaques in Alzheimer's disease brain, has been shown previously to undergo a process of polymerization to produce neurotoxic forms of amyloid. Recent literature has attempted to define precisely the form of Aβ responsible for its neurodegenerative properties. In the present study we describe a novel density-gradient centrifugation method for the isolation and characterization of structurally distinct polymerized forms of Aβ peptide. Fractions containing protofibrils, fibrils, sheet structures and low molecular mass oligomers were prepared. The fractionated forms of Aβ were characterized structurally by transmission electron microscopy. The effects on cell viability of these fractions was determined in the B12 neuronal cell line and hippocampal neurons. Marked effects on cell viability in the cells were found to correspond to the presence of protofibrillar and fibrillar structures, but not to monomeric peptide or sheet-like structures of polymerized Aβ. Biological activity correlated with a positive reaction in an immunoassay that specifically detects protofibrillar and fibrillar Aβ; those fractions that were immunoassay negative had no effect on cell viability. These data suggest that the effect of Aβ on cell viability is not confined to a single conformational form but that both fibrillar and protofibrillar species have the potential to be active in this assay.

scholarly journals OCCURRENCE, ISOLATION, AND CHARACTERIZATION OF POLYRIBOSOMES IN YEAST

1967 ◽  
Vol 34 (2) ◽  
pp. 505-512 ◽  
Author(s):  
Leon Marcus ◽  
H. Ris ◽  
H. O. Halvorson ◽  
R. K. Bretthauer ◽  
R. M. Bock
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Nearest Neighbors ◽  
Amino Acid Incorporation ◽  
Size Classes ◽  
Isolation And Characterization ◽  
Acid Incorporation

This report details the procedural requirements for preparing cell-free extracts of yeast rich in polyribosomes. This enabled us to demonstrate the occurrence of polyribosomes in yeast, to show their role in protein synthesis, and to devise methods for their resolution and isolation. When certain precautions are met (the use of log phase cells, rapidly halting cell growth, gentle methods of disruption, sedimentation through exponential density gradients, etc.), individual polyribosome size classes ranging up to the heptosome can be fractionated and separated from their nearest neighbors. Larger size classes are resolved partially among themselves, free of smaller polyribosomes. This was confirmed by extensive electron micrographic studies of material from the various fractions obtained upon density gradient centrifugation of yeast extracts. Modifications of the gradients and procedure should allow fractionation and isolation of the larger polyribosomes, including those containing polycistronic messages. Yeast polyribosomes are disaggregated to single ribosomes by longer term grinding, cell disruption by the French pressure cell, the Hughes press, or by incubation with dilute RNAse. Yeast polyribosomes are active in the incorporation of amino acids into polypeptide; the single ribosomes exhibit only slight activity. The latter activity is probably due to the presence of a small fraction of monosomes still containing mRNA. Poly-U stimulates amino acid incorporation only in the single ribosomes.

scholarly journals Isolation and characterization of live yeast cells from ancient vessels as a tool in bio-archeology

2019 ◽  
Author(s):  
Tzemach Aouizerat ◽  
Itai Gutman ◽  
Yitzhak Paz ◽  
Aren M. Maeir ◽  
Yuval Gadot ◽  
...  
Keyword(s):  
Ancient Dna ◽  
Residue Analysis ◽  
Genomic Analysis ◽  
Fermented Food ◽  
Yeast Cells ◽  
Fermented Beverages ◽  
Isolation And Characterization ◽  
Novel Approach ◽  
Experimental Archeology

AbstractAncient fermented food has been studied based on recipes, residue analysis and ancient-DNA techniques and reconstructed using modern domesticated yeast. Here, we present a novel approach. We hypothesize that enriched yeast populations in fermented beverages could have become the dominant species in storage vessels and the descendants of these yeast could be isolated and studied long after. To this end, using a pipeline of yeast isolation from clay vessels developed here, we screened for yeast cells in beverage-related and non-related ancient vessels and sediments, from several archeological sites. We found that yeast cells could be successfully isolated specifically from clay containers of fermented beverages. Genomic analysis revealed that these yeast are similar to those found in traditional African beverages. Phenotypically, they grow similar to modern-beer producing yeast. Both strongly suggesting that they are descendants of the original fermenting yeast. These findings provide modern microorganisms as a new tool in bio-archeology.ImportanceSo far, most of the study of ancient organisms was based mainly on the analysis of ancient DNA. Here we show that it is possible to isolate and study microorganisms, yeast in this case, from thousands of years old clay vessels, used for fermentation. We demonstrate that it is highly likely that these cells are descendants of the original yeast strains which participated in the fermentation process and were absorbed into the pottery vessels. Moreover, we characterize the isolated yeast their genome and the beer they produce. These results open new and exciting avenues in the study of domesticated microorganisms and contribute significantly to the fields of bio and experimental –archeology that aims to reconstruct ancient artifacts and products.

scholarly journals Isolation and characterization of human cervical-mucus glycoproteins

1983 ◽  
Vol 211 (1) ◽  
pp. 13-22 ◽  
Author(s):  
I Carlstedt ◽  
H Lindgren ◽  
J K Sheehan ◽  
U Ulmsten ◽  
L Wingerup
Keyword(s):  
Density Gradient ◽  
Proteinase Inhibitors ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Disc Electrophoresis ◽  
Guanidinium Chloride ◽  
Molecular Weights ◽  
Isolation And Characterization ◽  
Mucus Glycoproteins

Mucus glycoproteins (mucins) were extracted from human cervical pregnancy mucus by 6 M-guanidinium chloride in the presence of proteinase inhibitors. Purification was subsequently achieved by isopycnic density-gradient centrifugation in CsCl/ guanidinium chloride gradients. The purified macromolecules represented approx. 85% of the total and were devoid of nucleic acids and proteins, as judged by analytical density-gradient centrifugation, disc electrophoresis and u.v. spectroscopy. Sedimentation-velocity centrifugation revealed a single unimodal peak with S20,W 50.1S in 0.2M-NaCl and 37.0S in 6 M-guanidinium chloride. Molecular weights obtained by light-scattering were 9.7 × 10(6) and 5.9 × 10(6) in 0.2M-NaCl and 6 M-guanidinium chloride respectively. The chemical analyses were typical of those of epithelial mucins. The macromolecules contained approx. 20% (w/w) of protein, and 65% (w/w) was accounted for as carbohydrate. Serine and threonine constituted 32 mol/100 mol and proline 10 mol/100 mol of the amino acids. The major sugars found were N-acetylglucosamine (12.8%), N-acetylgalactosamine (9.7%), galactose (18.7%), sialic acid (15.0%) and fucose (7.5%).

Isolation and Characterization of Pseudomonas sp. STM 997 from Soil Sample having Potentiality to Degrade 3,6-Dimethyl-1-keto-1,2,3,4-tetrahydrocarbazole: A Novel Approach

2012 ◽  
Vol 168 (7) ◽  
pp. 1765-1777
Author(s):  
Biswanath Chakraborty ◽  
Suchandra Chakraborty ◽  
Anjan Kumar Basu ◽  
Bhrigu Aditya ◽  
T. P. Sinha ◽  
...  
Keyword(s):  
Soil Sample ◽  
Pseudomonas Sp ◽  
Isolation And Characterization ◽  
Novel Approach
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