scholarly journals Extraction and Characterization of Lipopolysaccharide from Salmonella typhi

2021 ◽  
pp. 236-242
Author(s):  
Nilima Patel
Keyword(s):  
Carbohydrate Chain ◽  
Extraction Methods ◽  
Salmonella Typhi ◽  
Gram Negative Bacteria ◽  
Ladder Pattern ◽  
Wet Weight ◽  
Phenol Water ◽  
Phenolic Extraction ◽  
Relative Purity

There is no standard process available for high purity LPS isolation, and it may be appropriate to mix two purification steps. For Salmonella typhi LPS that were subjected to normal microbiological & biochemical screening protocols, updated phenol-water extraction protocol & non-phenolic extraction methods were also used in present research. Pellet of crude LPS obtained with wet weight was 2.0 gm. yield of LPS was more by hot phenol method 1.94 mg/ml as compared to non-phenolic method 0.40 mg/ml. relative purity of LPS obtained by hot phenol method was more as compared to non-phenolic method, as protein content was 23.60 mg/ml in LPS extracted from hot phenol method & 26.62 mg/ ml in LPS extracted from non-phenolic method. However, Nucleic acid contain was comparable in LPS extracted from both methods. Qualitative analysis showed ladder like bands of LPS extracted by hot phenol method as compared to single band obtained for LPS extracted by non-phenolic method. Findings of silver staining clearly revealed ladder pattern of multi-rung bands that are characteristics of smooth form of gram negative bacteria due to difference in length of carbohydrate chain of O-antigen component. In order to clarify diseased conditions & better work on LPS profiling, this research may be crucial to lead to more successful diagnosis and care.

Characterization of dissolved organic matter (DOM) extracted from soils by hot water percolation (HWP)

2010 ◽  
Vol 59 (1) ◽  
pp. 99-108 ◽  
Author(s):  
M. Takács ◽  
Gy. Füleky
Keyword(s):  
Organic Matter ◽  
Humic Acid ◽  
Natural Organic Matter ◽  
Extraction Methods ◽  
Soil Samples ◽  
Hot Water ◽  
Soil Extracts ◽  
Water Percolation ◽  
Soil Humic Acid

The Hot Water Percolation (HWP) technique for preparing soil extracts has several advantages: it is easily carried out, fast, and several parameters can be measured from the same solution. The object of this study was to examine the possible use of HWP extracts for the characterization of soil organic matter. The HPLC-SEC chromatograms, UV-VIS and fluorescence properties of the HWP extracts were studied and the results were compared with those of the International Humic Substances Society (IHSS) Soil Humic Acid (HA), IHSS Soil Fulvic Acid (FA) and IHSS Suwannee Natural Organic Matter (NOM) standards as well as their HA counterparts isolated by traditional extraction methods from the original soil samples. The DOM of the HWP solution is probably a mixture of organic materials, which have some characteristics similar to the Soil FA fractions and NOM. The HWP extracted organic material can be studied and characterized using simple techniques, like UV-VIS and fluorescence spectroscopy.

Electrophysiological Characterization of Transport Across Outer Membrane Channels from Gram-Negative Bacteria in Their Native Environment

2019 ◽  
Author(s):  
Jiajun Wang ◽  
Rémi Terrasse ◽  
Jayesh Arun Bafna ◽  
Lorraine Benier ◽  
Mathias Winterhalter
Keyword(s):  
Outer Membrane ◽  
Lipid Membrane ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Multi Drug Resistance ◽  
Gram Negative Bacteria ◽  
Membrane Channels ◽  
Gram Negative ◽  
Electrophysiological Characterization

Multi-drug resistance in Gram-negative bacteria is often associated with low permeability of the outer membrane. To investigate the role of membrane channels in the uptake of antibiotics, we extract, purify and reconstitute them into artificial planar membranes. To avoid this time-consuming procedure, here we show a robust approach using fusion of native outer membrane vesicles (OMV) into planar lipid bilayer which moreover allows also to some extend the characterization of membrane protein channels in their native environment. Two major membrane channels from <i>Escherichia coli</i>, OmpF and OmpC, were overexpressed from the host and the corresponding OMVs were collected. Each OMV fusion revealed surprisingly single or only few channel activities. The asymmetry of the OMV´s translates after fusion into the lipid membrane with the LPS dominantly present at the side of OMV addition. Compared to conventional reconstitution methods, the channels fused from OMVs containing LPS have similar conductance but a much broader distribution. The addition of Enrofloxacin on the LPS side yields somewhat higher association (<i>k<sub>on</sub></i>) and lower dissociation (<i>k<sub>off</sub></i>) rates compared to LPS-free reconstitution. We conclude that using outer membrane vesicles is a fast and easy approach for functional and structural studies of membrane channels in the native membrane.

scholarly journals Synthesis, Characterization of Chitosan-Aluminum Oxide Nanocomposite for Green Synthesis of Annulated Imidazopyrazol Thione Derivatives

Polymers ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1160
Author(s):  
Abir S. Abdel-Naby ◽  
Sara Nabil ◽  
Sarah Aldulaijan ◽  
Ibtisam M. Ababutain ◽  
Azzah I. Alghamdi ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Aluminum Oxide ◽  
Reaction Medium ◽  
Thiol Group ◽  
Docking Studies ◽  
Significant Loss ◽  
Gram Negative Bacteria ◽  
Organic Catalyst ◽  
Group 1

Chitosan-aluminum oxide nanocomposite was synthesized, characterized, and used as a green heterogeneous catalyst to synthesize novel imidazopyrazolylthione derivatives. Nanocomposite polymeric material was characterized by EDS-SEM and XRD. The powerful catalytic activity, and its base character of the nanocomposite, was used to synthesize imidazopyrazolylthione (1) in a good yield compared to traditional cyclocondensation synthesis. Using the nanocomposite catalyst, substitution of the thiol group (1) afforded the corresponding thiourea (2) and the corresponding ester (3). The efficiency of the nanocomposite over the traditional base organic catalyst, Et3N and NaOH, makes it an effective, economic, and reproducible nontoxic catalyst. Moreover, the heterogeneous nanocomposite polymeric film was easily isolated from the reaction medium, and recycled up to four times, without a significant loss of its catalytic activity. The newly synthesized derivatives were screened as antibacterial agents and showed high potency. Molecular docking was also performed for a more in-depth investigation. The results of the docking studies have demonstrated that the docked compounds have strong interaction energies with both Gram-positive and Gram-negative bacteria.

scholarly journals Facile Synthesis and Characterization of Palm CNF-ZnO Nanocomposites with Antibacterial and Reinforcing Properties

2021 ◽  
Vol 22 (11) ◽  
pp. 5781
Author(s):  
Janarthanan Supramaniam ◽  
Darren Yi Sern Low ◽  
See Kiat Wong ◽  
Loh Teng Hern Tan ◽  
Bey Fen Leo ◽  
...  
Keyword(s):  
Plant Biomass ◽  
Cellulose Nanofibers ◽  
High Strength ◽  
Salmonella Typhi ◽  
Particle Sizes ◽  
Mechanical Reinforcement ◽  
Uniform Size ◽  
Preparation Methods ◽  
Potential Applications

Cellulose nanofibers (CNF) isolated from plant biomass have attracted considerable interests in polymer engineering. The limitations associated with CNF-based nanocomposites are often linked to the time-consuming preparation methods and lack of desired surface functionalities. Herein, we demonstrate the feasibility of preparing a multifunctional CNF-zinc oxide (CNF-ZnO) nanocomposite with dual antibacterial and reinforcing properties via a facile and efficient ultrasound route. We characterized and examined the antibacterial and mechanical reinforcement performances of our ultrasonically induced nanocomposite. Based on our electron microscopy analyses, the ZnO deposited onto the nanofibrous network had a flake-like morphology with particle sizes ranging between 21 to 34 nm. pH levels between 8–10 led to the formation of ultrafine ZnO particles with a uniform size distribution. The resultant CNF-ZnO composite showed improved thermal stability compared to pure CNF. The composite showed potent inhibitory activities against Gram-positive (methicillin-resistant Staphylococcus aureus (MRSA)) and Gram-negative Salmonella typhi (S. typhi) bacteria. A CNF-ZnO-reinforced natural rubber (NR/CNF-ZnO) composite film, which was produced via latex mixing and casting methods, exhibited up to 42% improvement in tensile strength compared with the neat NR. The findings of this study suggest that ultrasonically-synthesized palm CNF-ZnO nanocomposites could find potential applications in the biomedical field and in the development of high strength rubber composites.

scholarly journals Optimization of extraction methods for characterization of phenolic compounds in apricot fruit (Prunus armeniaca)

2019 ◽  
Vol 10 (10) ◽  
pp. 6492-6502
Author(s):  
Lisard Iglesias-Carres ◽  
Anna Mas-Capdevila ◽  
Francisca Isabel Bravo ◽  
Cinta Bladé ◽  
Anna Arola-Arnal ◽  
...  
Keyword(s):  
Phenolic Compounds ◽  
Extraction Methods ◽  
Prunus Armeniaca ◽  
Health Promoting

Fruits are rich in phenolic compounds with health-promoting activities.

scholarly journals Discovery and Characterization of the Antimetabolite Action of Thioacetamide-Linked 1,2,3-Triazoles as Disruptors of Cysteine Biosynthesis in Gram-Negative Bacteria

2019 ◽  
Vol 6 (3) ◽  
pp. 467-478 ◽  
Author(s):  
Miranda J. Wallace ◽  
Suresh Dharuman ◽  
Dinesh M. Fernando ◽  
Stephanie M. Reeve ◽  
Clifford T. Gee ◽  
...  
Keyword(s):  
Gram Negative Bacteria ◽  
Cysteine Biosynthesis ◽  
Gram Negative

Susceptibility of Gram-negative bacteria to β-lactam antibiotics and rapid characterization of β-lactamase activity

1979 ◽  
Vol 2 (4) ◽  
pp. 239-244 ◽  
Author(s):  
David L. Gibbs ◽  
Clyde Thornsberry
Keyword(s):  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Rapid Characterization

Hybridization by Deoxycholate Treatment or by Biological Degradation of Bacterial Endotoxins Extracted from Smooth and Rough Strains

1970 ◽  
Vol 1 (1) ◽  
pp. 15-20
Author(s):  
L. Chedid ◽  
M. Parant ◽  
F. Parant ◽  
F. Peroux ◽  
J. J. Perez
Keyword(s):  
Sodium Deoxycholate ◽  
Salmonella Typhi ◽  
Side Chains ◽  
Biological Degradation ◽  
Specific Antibodies ◽  
Electrophoretic Migration ◽  
Bacterial Endotoxins ◽  
Phenol Water ◽  
Smooth Strain

An endotoxin extracted with phenol-water from a rough microorganism ( Salmonella typhi strain R 2 ) was hybridized with an endotoxin obtained by the Boivin technique from a smooth strain ( S. enteritidis Danysz) when they were mixed in the presence of sodium deoxycholate. These two toxic antigens could also be hybridized by incubation in citrated serum. With this new composite molecule, the presence of the hydrophilic side chains on the smooth moiety influenced the electrophoretic migration of the R antigen and greatly hindered the reactivity of the R sites with their specific antibodies.

Protein expression and extraction of hard-to-produce proteins in the periplasmic space of Escherichia coli v1

2021 ◽  
Author(s):  
Cristina Hernandez Rollan ◽  
Kristoffer Bach Falkenberg ◽  
Maja Rennig ◽  
Andreas Birk Bertelsen ◽  
Morten Norholm
Keyword(s):  
Protein Production ◽  
Expression System ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
E Coli ◽  
New Family ◽  
Lytic Polysaccharide Monooxygenases ◽  
Strain Bl21 ◽  
Polysaccharide Monooxygenases

E. coli is a gram-negative bacteria used mainly in academia and in some industrial scenarios, as a protein production workhorse. This is due to its ease of manipulation and the range of genetic tools available. This protocol describes how to express proteins in the periplasm E. coli with the strain BL21 (DE3) using a T7 expression system. Specifically, it describes a series of steps and tips to express "hard-to-express" proteins in E. coli, as for instance, LPMOs. The protocol is adapted from Hemsworth, G. R., Henrissat, B., Davies, G. J., and Walton, P. H. (2014) Discovery and characterization of a new family of lytic polysaccharide monooxygenases. Nat. Chem. Biol.10, 122–126. .

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