scholarly journals Isolation and characterization of BpL1, a broad acting lytic bacteriophage against Brucella

2017 ◽  
Author(s):  
Vimlesh Gupta ◽  
Hari Mohan Saxena
Keyword(s):  
Pasteurella Multocida ◽  
Uv Light ◽  
Brucella Melitensis ◽  
Sodium Dodecyl ◽  
Dairy Farm ◽  
Tail Length ◽  
Lytic Bacteriophage ◽  
Isolation And Characterization ◽  
Rnase Treatment

AbstractWe have isolated a new broad acting lytic brucellaphage (BpL1) from the sewage of a dairy farm. The phage lysed all the 12Brucella abortusfield isolates,B. abortusstrain 99 andBrucella melitensisbut did not lyse any of the heterologous species tested viz.Staphylococcus aureus, Pasteurella multocida, Escherichia coli,andSalmonella species. Streaking the plaques onBrucellalawn gave clear zones along the streak lines. The plaques were circular with a diameter of 0.5- 3.0 mm. At a concentration of 10−4the phage count was 4.5 × 106plaques per ml. It was a tailed phage with icosahedral head (62.2 nm in diameter and 73.71 nm in length), and the head to tail length was 229.21 nm. The phage belonged to the order Caudovirales and familySiphoviridae. It was inactivated within one hour at 55°C and within 4 hours at −20°C. Treatment at pH 2 for 4 hours and at pH 4 for 12 hours inactivated it. It was inactivated after 4 hours exposure to sunlight, and within 4 minutes by UV light. Chloroform and Sodium Dodecyl Sulfate inactivated it within 15 minutes. Lysozyme inactivated it within 1 hour whereas RNase treatment did not affect its activity.

Preparative isolation and characterization of the B875 complex from Rhodobacter sphaeroides 2.4.1

1988 ◽  
Vol 66 (5) ◽  
pp. 442-448 ◽  
Author(s):  
Rafael Picorel ◽  
Gabriel Gingras
Keyword(s):  
Rhodobacter Sphaeroides ◽  
Phosphatidyl Ethanolamine ◽  
Sodium Dodecyl ◽  
Pigment Analysis ◽  
Preparative Isolation ◽  
Ethanolamine Phosphatidyl ◽  
Isolation And Characterization ◽  
Detergent System ◽  
Triton X

We have developed a simple and efficient method, using a mixed detergent system of sodium dodecyl sulfate and Triton X-100, for the preparative isolation of theB875 complex from Rhodobacter sphaeroides 2.4.1. As a bonus, the method allows the preparation of both the B875 and B800-850 complexes from the same batch of chromatophores. The preparations are spectrally pure, as indicated by absorption and circular dichroism spectroscopy. The latter method suggests that the Qy band of the B875 complex is due to weakly interacting bacteriochlorophyll molecules. Protein and pigment analysis shows that the B875 complex contains 2 mol of bacteriochlorophyll and 2 mol of sphaeroidene per mol of apoprotein (12 266 g), whereas the B800-850 complex contains 3 mol of bacteriochlorophyll and 1 mol of sphaeroidene per mol of apoprotein (11 497 g). While these stoichiometries are in accord with currently accepted models, they disagree with their published experimental basis. Phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, and diphosphatidyl glycerol were found to be present in the B875 complex.

scholarly journals Isolation and characterization of bovine alphaherpesvirus 2 strain from an outbreak of bovine herpetic mammillitis in a dairy farm

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Gianvito Lanave ◽  
Vittorio Larocca ◽  
Michele Losurdo ◽  
Cristiana Catella ◽  
Paolo Capozza ◽  
...  
Keyword(s):  
Dairy Farm ◽  
Isolation And Characterization

scholarly journals ISOLATION AND CHARACTERIZATION OF CARBENDAZIM DEGRADING Trichoderma harzianum RIFAI MUTANTS

2010 ◽  
Vol 20 ◽  
Author(s):  
ITAMAR SOARES DE MELO ◽  
CÉLIA MARIA MAGANHOTTO DE S. SILVA ◽  
JANE L. FAULL
Keyword(s):  
Trichoderma Harzianum ◽  
Uv Light ◽  
Light Exposure ◽  
Wild Type ◽  
Pesticide Degradation ◽  
Isolation And Characterization ◽  
Degradation Ability

Mutants of Trichoderma harzianum Rifai, obtained after ultraviolet (UV) light exposure, showed high resistant to the fungicide benomyl. A mutant (2B6) was capable of degrading carbendazim, other fungicide of the benzimidazole fungicide. This mutant degraded 41.5% of the molecule within five days. This and others mutants (2B1 and 2B2) presented variation in size and frequency of uni-nucleated and/or bi-nucleated spores compared to the wild type. Four primers generated RAPDs patterns that allowed the mutant to be differentiated from the wild-type. It is concluded that using UV mutagenization, it is feasible to obtain strains of T. harzianum with improved pesticide degradation ability.

scholarly journals Isolation and characterization of a lytic bacteriophage (VB_PAnP_PADP4) against MDR- Pseudomonas aeruginosa isolated from septic wound infections

2019 ◽  
Vol 18 (15) ◽  
pp. 325-333
Author(s):  
Rani Pallavali Roja ◽  
Lakshmi Degati Vijaya ◽  
Venkata Rami Reddy Narala ◽  
Raghava Prasad Durbaka Vijaya
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Lytic Bacteriophage ◽  
Wound Infections ◽  
Isolation And Characterization ◽  
Septic Wound

scholarly journals Isolation and characterization of a lytic bacteriophage φKp-lyy15 of Klebsiella pneumoniae

2015 ◽  
Vol 30 (1) ◽  
pp. 66-68 ◽  
Author(s):  
Yinyin Lu ◽  
Hongyan Shi ◽  
Zhe Zhang ◽  
Fang Han ◽  
Jinghua Li ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Lytic Bacteriophage ◽  
Isolation And Characterization

Isolation and characterization of a FAD-dependent NADH diaphorase from Methanospirillum hungatei strain GP1

1981 ◽  
Vol 59 (2) ◽  
pp. 83-91 ◽  
Author(s):  
R. C. McKellar ◽  
K. M. Shaw ◽  
G. D. Sprott
Keyword(s):  
Gel Electrophoresis ◽  
Superoxide Anion ◽  
Chelating Agents ◽  
Sodium Dodecyl ◽  
Sulfhydryl Reagents ◽  
Ph Optimum ◽  
Methanospirillum Hungatei ◽  
Isolation And Characterization ◽  
Nadh Diaphorase

Crude extracts of Methanospirillum hungatei strain GP1 contained NADH and NADPH diaphorase activities. After a 483-fold purification of the NADH diaphorase the enzyme was further separated from contaminating proteins by polyacrylamide disc gel electrophoresis. Two distinct activity bands were extracted from the acrylamide, each one having oxygen, 2,6-dichlorophenoiindophenol, and cytochrome c linked activities. In these preparations NADPH could not replace NADH as electron donor. During the initial purification steps all activity was lost due to the removal of a readily released cofactor. Enzyme activity was restored by either FAD or a FAD fraction isolated from M. hungatei. Oxidase activity exhibited a broad pH optimum from 7.0 to 8.5 and apparent Km values of 26 μM for NADH and 0.2 μM for FAD. Superoxide anion, formed in the presence of oxygen, accounted for all of the NADH consumed in this reaction. The molecular weight of the diaphorase was about 117 500 by sodium dodecyl sulfate gel electrophoresis. Sulfhydryl reagents and chelating agents were inhibitory. Inactivation, which occurred during storage in phosphate buffer at 4 °C, was delayed by dithiothreitol. The isolated NADH diaphorase lacked NADPH:NAD transhydrogenase and NAD reductase activities.

scholarly journals Isolation and characterization of the urothelial lumenal plasma membrane.

1977 ◽  
Vol 73 (2) ◽  
pp. 382-399 ◽  
Author(s):  
J S Caruthers ◽  
M A Bonneville
Keyword(s):  
Plasma Membrane ◽  
Sodium Dodecyl ◽  
Modified Technique ◽  
Sialic Acid Content ◽  
Polarity Index ◽  
Total Sialic Acid ◽  
Isolation And Characterization ◽  
A Value

The lumenal plasma membrane has been isolated from transitional epithelial cells (urothelium) lining the urinary bladder in sheep by a modified technique involving treatment with hypotonic thioglycolate. The isolated membranes, like those in situ, are distinguished morphologically by arrays of hexagonal particles (in plague regions) separated by smooth interplaque regions. These plaque regions, specifically, can be isolated from the lumenal plasma membrane. Of the proteins constituting the lumenal plasma membrane, five were found to characterize the plaque regions and, in particular, the 33,000-dalton species appears to be most heavily concentrated in the sodium dodecyl sulfate-polyacrylamide gel pattern of the isolated plaque regions. Lipid analyses showed that there are approximately 0.93 mg of phospholipid and 0.27 mg of cholesterol for each milligram of protein, giving a value of 55% lipids and 45% proteins for the composition of the lumenal plasma membrane. The total sialic acid content was measured to be approximately 0.038 micronmol/mg protein for the plasma membrane. Several plasma membrane marker enzymes were found to be associated with the lumenal plasma membrane fraction, but only the 5'-nucleotidase activity was found to be further enriched in the plaque region fraction. Amino acid analysis of the intrinsic proteins of the plaques indicated a polarity index of 45%.

scholarly journals Isolation and characterization of human plasma α 1-proteinase inhibitor and a conformational study of its interaction with proteinases

1976 ◽  
Vol 157 (2) ◽  
pp. 339-351 ◽  
Author(s):  
J Saklatvala ◽  
G C Wood ◽  
D D White
Keyword(s):  
Human Plasma ◽  
Isoelectric Focusing ◽  
Proteinase Inhibitor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sedimentation Equilibrium ◽  
Isolation And Characterization ◽  
Step Procedure ◽  
Helical Content

1. alpha 1-Proteinase inhibitor was isolated from human plasma by a five-step procedure. Isoelectric focusing showed that six components focused between pH4.85 and 4.95. 2. The mol.wt. of the inhibitor was 52000 by sedimentation equilibrium and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The amino acid and carbohydrate compositions of the inhibitor were also determined. 3. The far-u.v.c.d. (circular-dichroism) spectrum indicated that the inhibitor had about 36% alpha-helical content. 4. The loss of proteinase-inhibitory activity when the inhibitor was exposed to pH values less than 5.0 or greater than 10.5 was accompanied by small changes in the far-u.v.c.d. spectrum and large changes in the near-u.v.c.d. spectrum. The change at alkaline pH was associated with ionization of tyrosine residues. 5. Interaction of inhibitor with chymotrypsin caused perturbation of the c.d. spectrum and this was used to follow the interaction and show a 1:1 stoicheiometry. 6. C.d., electrophoresis and isoelectric focusing showed that the inhibitor-enzyme complex is degraded by free enzyme. 7. Parallel studies with trypsin indicated that it too forms a 1:1 complex with inhibitor and is degraded by excess of enzyme.

Glycosylation of chicken haptoglobin: Isolation and characterization of three molecular variants and studies of their distribution in hen plasma before and after turpentine-induced inflammation

1988 ◽  
Vol 66 (3) ◽  
pp. 208-217 ◽  
Author(s):  
Francisco Delers ◽  
Gérard Strecker ◽  
Robert Engler
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Isolation And Characterization ◽  
Molecular Variants ◽  
Hemoglobin Binding ◽  
Significant Difference ◽  
Before And After ◽  
Carbohydrate Unit

Chicken haptoglobin (Hp), a hemoglobin-binding protein isolated from chicken plasma, is composed of three molecular variants that react differently with concanavalin A (ConA). These glycosylation variants of chicken Hp have been isolated by affinity chromatography using Sepharose-bound ConA. They differ in their molecular weight, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Analysis of the glycopeptides obtained after pronase digestion of these variants yielded two types of structures: one, reactive with ConA, corresponded to a biantennary N-linked carbohydrate unit and one, unreactive with ConA, corresponded to a triantennary unit. The strongly ConA-reactive Hp variant bears only two biantennary units and the nonreactive Hp variant bears only two triantennary units; the weakly reactive Hp variant bears equal amounts of both units. The distribution of Hp glycosylation variant does not show any significant difference when obtained from the plasma of laying hens before and after turpentine-induced inflammation.

The Isolation and Characterization of the ATPase Inhibitory Protein (TN-I) from Bovine Cardiac Muscle

1975 ◽  
Vol 53 (11) ◽  
pp. 1207-1213 ◽  
Author(s):  
L. D. Burtnick ◽  
W. D. McCubbin ◽  
C. M. Kay
Keyword(s):  
Molecular Weight ◽  
Cardiac Muscle ◽  
Calcium Binding ◽  
Sodium Dodecyl ◽  
Basic Amino Acid ◽  
Sedimentation Equilibrium ◽  
Amino Acid Residues ◽  
Inhibitory Protein ◽  
Isolation And Characterization

The inhibitory component of the troponin complex (TN-I) was purified from bovine cardiac muscle, using a combination of ion exchange and molecular exclusion chromatographies in the presence of urea. It has the ability to inhibit the Mg2+-activated ATPase (EC 3.6.1.3) of a synthetic cardiac actomyosin preparation and this inhibition is reversed by the addition of cardiac calcium binding component of troponin (TN-C). Conventional sedimentation equilibrium experiments suggest a molecular weight for cardiac TN-I of 22 900 ± 500. However, sodium dodecyl sulfate (SDS) gels indicate a molecular weight of 27 000 ± 1000. The mobility of TN-I on SDS gels may be anomalous due to the high proportion of basic amino acid residues in the protein. Cardiac TN-I and TN-C interact to form a tight complex, even in the presence of 6 M urea. The results of this study invite direct comparison with results published for rabbit skeletal TN-I.

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