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1.
Arch Microbiol ; 206(5): 234, 2024 Apr 25.
Article in English | MEDLINE | ID: mdl-38664262

ABSTRACT

Exploration and marketable exploitation of coalbed methane (CBM) as cleaner fuel has been started globally. In addition, incidence of methane in coal basins is an imperative fraction of global carbon cycle. Significantly, subsurface coal ecosystem contains methane forming archaea. There is a rising attention in optimizing microbial coal gasification to exploit the abundant or inexpensive coal reserves worldwide. Therefore, it is essential to understand the coalbeds in geo-microbial perspective. Current review provides an in-depth analysis of recent advances in our understanding of how methanoarchaea are distributed in coal deposits globally. Specially, we highlight the findings on coal-associated methanoarchaeal existence, abundance, diversity, metabolic activity, and biogeography in diverse coal basins worldwide. Growing evidences indicates that we have arrived an exciting era of archaeal research. Moreover, gasification of coal into methane by utilizing microbial methanogenesis is a considerable way to mitigate the energy crisis for the rising world population.


Subject(s)
Archaea , Coal , Methane , Methane/metabolism , Archaea/metabolism , Archaea/genetics , Ecosystem , Phylogeny
2.
Int J Biol Macromol ; 253(Pt 4): 127004, 2023 Dec 31.
Article in English | MEDLINE | ID: mdl-37734526

ABSTRACT

A thoughtful strategy has been intended to control the hydrogel networking to assess the binding efficacy of multifunctional hydrogel. The processing of two distinct network-supported hydrogels has portrayed to express the operating interactions involved during co-existence with solvents, small molecules, biomolecules, etc. Herein, chitosan has separately functionalized in semisynthetic approaches with 4-hydroxyisopthalaldehyde (ChDA) and 2-hydroxybenzene-1,3,5-tricarbaldehyde (ChTA) to construct different gel networks. The disposition of gel networks ChDA adapts more flexible chain or spine, whereas ChTA possesses restricted movements within gel networks. The gel networks of hydrogels have a significant role in their distinct physical activities. Their gel-bonding elucidations have performed to establish the variation in mechanical, swelling photophysical properties, etc. Remarkable self-fluorescence behaviors are used as a tool for binding study. Distinctive gel networks and their flexibility have investigated against self-fluorescence, UV-Vis, and FTIR against small molecule, Boron trifluoride and biomolecule, and Bovine serum albumin. Hydrogel/BF3 shows variation in fluorescence due to the disposition of gel networks. Hydrogel/BSA quenching of fluorescence at three different temperatures provides the binding constant and Stern-Volmer quenching constant. Theoretical DFT and docking studies successfully established the flexibility against binding study. The controlling of cross-linking or functionalization is very crucial for the development of hydrogel-mediated applications.


Subject(s)
Chitosan , Chitosan/chemistry , Hydrogels/chemistry , Serum Albumin, Bovine/chemistry
3.
Carbohydr Polym ; 278: 118965, 2022 Feb 15.
Article in English | MEDLINE | ID: mdl-34973780

ABSTRACT

Utilization of biomolecules encapsulated nano particles is currently originating ample attention to generate unconventional nanomedicines in antiviral research. Zinc oxide nanoparticle has been extensively studied for antimicrobial, antifungal and antifouling properties due to high surface to volume ratios and distinctive chemical as well as physical properties. Nevertheless, still minute information is available on their response on viruses. Here, in situ nanostructured and polysaccharide encapsulated ZnO NPs are fabricated with having antiviral potency and low cytotoxicity (%viability ~ 90%) by simply controlling the formation within interspatial 3D networks of hydrogels through perfect locking mechanism. The two composites ChH@ZnO and ChB@ZnO shows exceedingly effective antiviral activity toward Human cytomegalovirus (HCMV) having cell viability 93.6% and 92.4% up to 400 µg mL-1 concentration. This study brings significant insights regarding the role of ZnO NPs surface coatings on their nanotoxicity and antiviral action and could potentially guide to the development of better antiviral drug.


Subject(s)
Antiviral Agents/pharmacology , Benzaldehydes/pharmacology , Chitosan/pharmacology , Cytomegalovirus/drug effects , Nanoparticles/chemistry , Zinc Oxide/pharmacology , Antiviral Agents/chemistry , Benzaldehydes/chemistry , Chitosan/chemistry , Humans , Microbial Sensitivity Tests , Molecular Structure , Zinc Oxide/chemistry
4.
Int J Biol Macromol ; 193(Pt A): 758-767, 2021 Dec 15.
Article in English | MEDLINE | ID: mdl-34717978

ABSTRACT

Hydrogels are considered as practical and proficient materials in adsorption and removal of soluble lethal molecules from aqueous system. They are also rapid-decomposable and economical materials besides their diverse preventive claims. In current study, Cinnamaldehyde (C), a natural defensive compound and Chitosan (Ch), natural occurring bio-macromolecule are considered to develop bio-inspired hydrogel (ChC). The structural and surface characteristics of ChC (13C solid state NMR, FT-IR, UV-Vis and SEM) are investigated to confirm the successful grafting. The origami of gelation in ChC performs an excellent adsorption activity towards food dyes, Carmoisine (CA) and Tartrazine (TA), which are contaminated by the accumulation during excess release from catering and chemical industries in aqueous system. The adsorption performance is thoroughly screened by varying the pH, ChC dosage, dye concentration, contact time and temperature in aqueous system. Thermodynamic and Kinetics study suggest the natural tendency of adsorption with a good reusability up to 3 cycles. The main mechanism for spontaneous adsorption is initiated by capturing of TA/CA in porous surface followed by the ionic interactions and formation of H-bondings. ChC based adsorption is an excellent and potential approach to control the toxicants for the water-pollution and water-preservation.


Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Chitosan/chemistry , Color , Hydrogels/chemistry , Water/chemistry , Acetic Acid , Acrolein/analogs & derivatives , Adsorption , Coloring Agents/chemistry , Hydrogen-Ion Concentration , Kinetics , Naphthalenesulfonates , Spectroscopy, Fourier Transform Infrared , Tartrazine , Thermodynamics , Water Pollution
5.
Parasite Immunol ; 43(3): e12806, 2021 03.
Article in English | MEDLINE | ID: mdl-33131110

ABSTRACT

The anti-leishmanial effect of the 'carbohydrate-fraction', isolated from an edible mushroom Astraeus hygrometricus, was evaluated against Leishmania donovani infection both in vitro and in vivo. Ahf-Car induced the expression of inducible nitric oxide synthase 2 (iNOS2) and pro-inflammatory cytokines like TNF-α and IL-12, with subsequent downregulation of the anti-inflammatory cytokines as TGF-ß and IL-10, in vitro and in vivo along with a remarkable increase in the expressions of IL-6, IL-1ß, IFN-γ and IRFs, IRF-7 and IRF-8 in vivo. Ahf-Car also reduced the parasite burden in the spleen and liver dose-dependently with a simultaneous proliferation of Ly6C+ cells in the bone marrow of Leishmania-infected experimental animals. It also increased the monocyte population dose-dependently and the expression of the myeloid transcription factor PU.1, in vivo, which presumably signifies the expansion of protective macrophages. Thus, Ahf-Car might be a potent anti-leishmanial lead with unique and effective adjuvant capacity.


Subject(s)
Basidiomycota/chemistry , Biological Products/therapeutic use , Leishmania donovani , Leishmaniasis, Visceral/prevention & control , Adjuvants, Immunologic/pharmacology , Animals , Biological Products/isolation & purification , Cytokines/immunology , Interleukin-12/immunology , Leishmania donovani/immunology , Leishmaniasis, Visceral/immunology , Liver/parasitology , Macrophages/immunology , Male , Mice , Mice, Inbred BALB C , Spleen/immunology , Spleen/parasitology , Tumor Necrosis Factor-alpha/immunology
6.
Carbohydr Polym ; 251: 116999, 2021 Jan 01.
Article in English | MEDLINE | ID: mdl-33142568

ABSTRACT

The fabrication of thiophene-chitosan (TCS) hydrogel has been carried out to show the excellent binding performance of Hg(II) from an aqueous solution of heavy metal ions in presence of thiophene moiety within the hydrogel network. Thiophene moiety has been implanted within chitosan, a wild bio-resources, through a facile Schiff base condensation strategy with 2-thiophenecarboxaldehyde to develop a three-dimensional network of TCS hydrogel. The parameters influencing adsorption capacity such as pH, volume of functional agent, contact time, amount of the hydrogel are included to broaden the in-depth study for the adsorption window of Hg(II) followed by the desorption and reusability performance of TCS. The results indicate that the TCS hydrogel for Hg(II) followed pseudo-second-order kinetics. Ethylenediaminetetraacetic acid (EDTA), acts as a better eluent compared to HCl to desorb Hg(II) and even after recurring adsorption/desorption cycles, removal efficacy of TCS hydrogel could be retained.


Subject(s)
Chitosan , Mercury/isolation & purification , Thiophenes , Water Pollutants, Chemical/isolation & purification , Adsorption , Cations, Divalent/isolation & purification , Chitosan/chemical synthesis , Chitosan/chemistry , Humans , Hydrogels , Hydrogen-Ion Concentration , Kinetics , Magnetic Resonance Spectroscopy , Microscopy, Electron , Molecular Structure , Rheology , Thiophenes/chemical synthesis , Thiophenes/chemistry , Water Purification/methods
7.
J Basic Microbiol ; 60(11-12): 938-949, 2020 Nov.
Article in English | MEDLINE | ID: mdl-33022819

ABSTRACT

The production of extracellular polysaccharides (EPS) by haloarchaeal members, with novel and unusual physicochemical properties, is of special importance and has the potential for extensive biotechnological exploitation. An extremely halophilic archaeon, Haloferax sp. BKW301 (GenBank Accession No. KT240044) isolated from a solar saltern of Baksal, West Bengal, India has been optimized for the production of EPS under batch culture. It produced a considerable amount (5.95 g/L) of EPS in the medium for halophiles with 15% NaCl, 3% glucose, 0.5% yeast extract, and 6% inoculum under shake flask culture at 120 rpm. The purified EPS, a homopolymer of galactose as revealed by chromatographic methods and Fourier-transform infrared spectroscopy, is noncrystalline (CIxrd , 0.82), amorphous, and could emulsify hydrocarbons like kerosene, petrol, xylene, and so forth. Moreover, the polymer is highly thermostable (up to 420°C) and displayed pseudoplastic rheology. Biologically, the EPS was able to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) radical efficiently and inhibit the proliferation of the Huh-7 cell line at an IC50 value of 6.25 µg/ml with a Hill coefficient of 0.844. Large-scale production of this thermostable, pseudoplastic homopolysaccharide, therefore, could find suitable applications in industry and biotechnology.


Subject(s)
Haloferax/metabolism , Polysaccharides, Bacterial/metabolism , Batch Cell Culture Techniques , Biopolymers/chemistry , Biopolymers/isolation & purification , Biopolymers/metabolism , Biopolymers/pharmacology , Cell Line, Tumor , Cell Proliferation/drug effects , Culture Media , Emulsifying Agents , Free Radical Scavengers , Galactose , Haloferax/classification , Haloferax/genetics , Hot Temperature , Humans , India , Phylogeny , Polysaccharides, Bacterial/chemistry , Polysaccharides, Bacterial/isolation & purification , Polysaccharides, Bacterial/pharmacology , Rheology
8.
Int J Biol Macromol ; 141: 626-635, 2019 Dec 01.
Article in English | MEDLINE | ID: mdl-31494166

ABSTRACT

Bio-resources have a very significant role in current research approach for the synthesis of benign functionalized biological macromolecules for their stable structural integrity and inherent nature-inspired potentialities. Here, chitosan is used as a core moiety for designing of a porous adsorbent after the attachment of salicylaldehyde to remove the toxic dyes. Salicylaldehyde linked chitosan, with excellent surface porosity, lightweight, non-glucose and low-cost feature, makes it as an efficient adsorbent. The dye loaded material is very easy to remove from the top of the water as it is suspended on water. The physico-chemical characterizations are done by FTIR, rheology, SEM and swelling study. The removal efficiency is 98% and 99% for Crystal Violet and Rose Bengal from water respectively. The thorough adsorption with mechanistic approach shows the Freundlich model as an appropriate one and follows closely pseudo-second-order kinetics model. Thermodynamic study reveals the endothermic nature of the process. Moreover, the reusability of Salicylaldehyde linked Chitosan shows its persistence with the same amount and concentration of dyes in water up to three consecutive cycles. So, the chitosan based macromolecules can be a sustainable candidate in the current scenario for the removal of dyes without the dislocation of the water container.


Subject(s)
Aldehydes/chemistry , Chitosan/chemistry , Coloring Agents/chemistry , Coloring Agents/isolation & purification , Adsorption , Hydrogen-Ion Concentration , Kinetics , Temperature
9.
Chemistry ; 25(18): 4856-4863, 2019 Mar 27.
Article in English | MEDLINE | ID: mdl-30693577

ABSTRACT

In the present study, the carbazole and 2,3,3-triphenylacrylonitrile (TPAN) nanostructures (2-CTPAN and 2,2'-CTPAN) have been designed and synthesized by Pd-catalyzed Sonogashira cross-coupling reaction. CTPAN exhibit aggregation-induced emission enhancement (AIEE) behavior in water with high fluorescence quantum yield. Both the compounds show tunable self-assembly in water as well as in N,N-dimethylformamide (DMF) by extended π-π stacking interactions. CTPAN can be self-assembled into spherical particles in water and the structures of these self-assemblies have been investigated using X-ray diffraction. Interestingly, 2-CTPAN and 2,2'-CTPAN form organogels with a critical gelation concentration (CGC) of 11 and 15 mg mL-1 , respectively, in DMF and exhibit acicular and rod shaped morphology, respectively. The single-crystal structure of 2-CTPAN shows that the intermolecular C-H⋅⋅⋅π interactions lock the molecular conformation into a staircase-shaped supramolecular assembly. These AIEE active compounds reveal high water dispersibility, strong yellow fluorescence with high quantum yield, promising photostability and excellent biocompatibility, which make them potential bioimaging agents.

10.
Carbohydr Polym ; 193: 119-128, 2018 Aug 01.
Article in English | MEDLINE | ID: mdl-29773363

ABSTRACT

A three-dimensional fluorescent hydrogel based on chitosan, polyvinyl alcohol and 9-anthraldehyde (ChPA) has been successfully designed and synthesized for the selective detection and discrimination of Fe3+ and Fe2+ in aqueous environment. The unique characteristics of ChPA has been confirmed by the Fourier-transform infrared spectroscopy (FTIR), rheological measurement, scanning electron microscopy (SEM), thermogravimetry and differential thermogravimetry (TG-DTG), ultraviolet-visible spectroscopy (UV-vis), fluorescence studies, transmission electron microscopy (TEM), energy dispersive x-ray spectroscopy (EDX), x-ray diffraction (XRD) and dynamic light scattering (DLS). The emission intensity at 516 nm of the hydrogel has been enhanced remarkably with the addition of Fe3+ due to the inhibition of the photoinduced electron transfer (PET) process. However, it gets strongly quenched in the case of Fe2+ owing to chelation enhanced quenching (CHEQ). The probe (ChPA) causes no significant change in the fluorescence and becomes highly specific and sensitive towards Fe3+ and Fe2+ compared to other interfering heavy and transition metal ions (HTM). The detection limits of the sensor for the Fe3+ and Fe2+ are 0.124 nM and 0.138 nM, respectively. The probe is also promising as a selective sensor for the Fe3+ and Fe2+ in the fluorescence imaging of living cells. Thus, such a probe opens up new opportunities to improve the chitosan based fluorescent chemosensor having biocompatibility, biodegradability, sufficient thermal stability and stability in a wide pH range.

11.
Chem Cent J ; 11(1): 118, 2017 Nov 21.
Article in English | MEDLINE | ID: mdl-29159657

ABSTRACT

Polysaccharides are structurally complex and essential constituents of life, and therefore, studies directed to these kinds of molecules have received scientific attention. Despite an easy availability of Dolichos biflorus Linn and Trachyspermum ammi (Linn) seeds isolation, characterization and antimicrobial studies of polysaccharides derived from these two natural sources have not been investigated. Therefore, we report here isolation of polysaccharides, their purification and characterization from Dolichos biflorus Linn and Trachyspermum ammi (Linn) seeds. Gel permeation chromatography, GC-MS, SEM, XRD, EDX and FT-IR analyses show the presence of three pentose sugar such as D-ribose, D-arabinose, D-xylose and hexose sugar such as D-mannose, D-galactose and D-glucose. Unprecedented antimicrobial activity of these polysaccharides against Gram positive bacteria such as Staphylococcus aureus and Bacillus subtilis and Gram negative bacteria such as Escherichia coli and Pseudomonas aeruginosa are established.

12.
PLoS One ; 12(8): e0183115, 2017.
Article in English | MEDLINE | ID: mdl-28832613

ABSTRACT

Bacillus anthracis (Ba) and human infection-associated Bacillus cereus (Bc) strains Bc G9241 and Bc 03BB87 have secondary cell wall polysaccharides (SCWPs) comprising an aminoglycosyl trisaccharide repeat: →4)-ß-d-ManpNAc-(1→4)-ß-d-GlcpNAc-(1→6)-α-d-GlcpNAc-(1→, substituted at GlcNAc residues with both α- and ß-Galp. In Bc G9241 and Bc 03BB87, an additional α-Galp is attached to O-3 of ManNAc. Using NMR spectroscopy, mass spectrometry and immunochemical methods, we compared these structures to SCWPs from Bc biovar anthracis strains isolated from great apes displaying "anthrax-like" symptoms in Cameroon (Bc CA) and Côte d'Ivoire (Bc CI). The SCWPs of Bc CA/CI contained the identical HexNAc trisaccharide backbone and Gal modifications found in Ba, together with the α-Gal-(1→3) substitution observed previously at ManNAc residues only in Bc G9241/03BB87. Interestingly, the great ape derived strains displayed a unique α-Gal-(1→3)-α-Gal-(1→3) disaccharide substitution at some ManNAc residues, a modification not found in any previously examined Ba or Bc strain. Immuno-analysis with specific polyclonal anti-Ba SCWP antiserum demonstrated a reactivity hierarchy: high reactivity with SCWPs from Ba 7702 and Ba Sterne 34F2, and Bc G9241 and Bc 03BB87; intermediate reactivity with SCWPs from Bc CI/CA; and low reactivity with the SCWPs from structurally distinct Ba CDC684 (a unique strain producing an SCWP lacking all Gal substitutions) and non-infection-associated Bc ATCC10987 and Bc 14579 SCWPs. Ba-specific monoclonal antibody EAII-6G6-2-3 demonstrated a 10-20 fold reduced reactivity to Bc G9241 and Bc 03BB87 SCWPs compared to Ba 7702/34F2, and low/undetectable reactivity to SCWPs from Bc CI, Bc CA, Ba CDC684, and non-infection-associated Bc strains. Our data indicate that the HexNAc motif is conserved among infection-associated Ba and Bc isolates (regardless of human or great ape origin), and that the number, positions and structures of Gal substitutions confer unique antigenic properties. The conservation of this structural motif could open a new diagnostic route in detection of pathogenic Bc strains.


Subject(s)
Bacillus anthracis/pathogenicity , Bacillus cereus/pathogenicity , Polysaccharides/metabolism , Animals , Bacillus anthracis/metabolism , Bacillus cereus/metabolism , Nuclear Magnetic Resonance, Biomolecular , Polysaccharides/chemistry , Primates , Rabbits
13.
PLoS One ; 9(9): e106513, 2014.
Article in English | MEDLINE | ID: mdl-25215579

ABSTRACT

The decoration of the lipid A headgroups of the lipooligosaccharide (LOS) by the LOS phosphoethanolamine (PEA) transferase (LptA) in Neisseria spp. is central for resistance to polymyxin. The structure of the globular domain of LptA shows that the protein has five disulphide bonds, indicating that it is a potential substrate of the protein oxidation pathway in the bacterial periplasm. When neisserial LptA was expressed in Escherichia coli in the presence of the oxidoreductase, EcDsbA, polymyxin resistance increased 30-fold. LptA decorated one position of the E. coli lipid A headgroups with PEA. In the absence of the EcDsbA, LptA was degraded in E. coli. Neisseria spp. express three oxidoreductases, DsbA1, DsbA2 and DsbA3, each of which appear to donate disulphide bonds to different targets. Inactivation of each oxidoreductase in N. meningitidis enhanced sensitivity to polymyxin with combinatorial mutants displaying an additive increase in sensitivity to polymyxin, indicating that the oxidoreductases were required for multiple pathways leading to polymyxin resistance. Correlates were sought between polymyxin sensitivity, LptA stability or activity and the presence of each of the neisserial oxidoreductases. Only meningococcal mutants lacking DsbA3 had a measurable decrease in the amount of PEA decoration on lipid A headgroups implying that LptA stability was supported by the presence of DsbA3 but did not require DsbA1/2 even though these oxidoreductases could oxidise the protein. This is the first indication that DsbA3 acts as an oxidoreductase in vivo and that multiple oxidoreductases may be involved in oxidising the one target in N. meningitidis. In conclusion, LptA is stabilised by disulphide bonds within the protein. This effect was more pronounced when neisserial LptA was expressed in E. coli than in N. meningitidis and may reflect that other factors in the neisserial periplasm have a role in LptA stability.


Subject(s)
Bacterial Proteins/metabolism , Drug Resistance, Bacterial/drug effects , Ethanolaminephosphotransferase/metabolism , Lipid A/metabolism , Neisseria meningitidis/enzymology , Oxidoreductases/metabolism , Polymyxins/pharmacology , Biocatalysis/drug effects , Disulfides/metabolism , Enzyme Stability/drug effects , Escherichia coli/metabolism , Lipopolysaccharides/pharmacology , Mutation/genetics , Neisseria meningitidis/drug effects , Oxidation-Reduction/drug effects , Periplasm/drug effects , Periplasm/metabolism
14.
Infect Immun ; 82(6): 2170-9, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24686069

ABSTRACT

The induction of an intense inflammatory response by Neisseria gonorrhoeae and the persistence of this pathogen in the presence of innate effectors is a fascinating aspect of gonorrhea. Phosphoethanolamine (PEA) decoration of lipid A increases gonococcal resistance to complement-mediated bacteriolysis and cationic antimicrobial peptides (CAMPs), and recently we reported that wild-type N. gonorrhoeae strain FA1090 has a survival advantage relative to a PEA transferase A (lptA) mutant in the human urethral-challenge and murine lower genital tract infection models. Here we tested the immunostimulatory role of this lipid A modification. Purified lipooligosaccharide (LOS) containing lipid A devoid of the PEA modification and an lptA mutant of strain FA19 induced significantly lower levels of NF-κB in human embryonic kidney Toll-like receptor 4 (TLR4) cells and murine embryonic fibroblasts than wild-type LOS of the parent strain. Moreover, vaginal proinflammatory cytokines and chemokines were not elevated in female mice infected with the isogenic lptA mutant, in contrast to mice infected with the wild-type and complemented lptA mutant bacteria. We also demonstrated that lptA mutant bacteria were more susceptible to human and murine cathelicidins due to increased binding by these peptides and that the differential induction of NF-κB by wild-type and unmodified lipid A was more pronounced in the presence of CAMPs. This work demonstrates that PEA decoration of lipid A plays both protective and immunostimulatory roles and that host-derived CAMPs may further reduce the capacity of PEA-deficient lipid A to interact with TLR4 during infection.


Subject(s)
Cathelicidins/pharmacology , Gonorrhea/immunology , Lipid A/chemistry , Neisseria gonorrhoeae/immunology , Reproductive Tract Infections/microbiology , Animals , Anti-Bacterial Agents/pharmacology , Cell Line, Transformed , Chemokines/metabolism , Complement System Proteins/immunology , Cytokines/metabolism , Ethanolamines , Female , Fibroblasts/drug effects , Gonorrhea/metabolism , Humans , Lipid A/immunology , Mice , Mice, Inbred BALB C , Microbial Sensitivity Tests , NF-kappa B/metabolism , Neisseria gonorrhoeae/drug effects , Neisseria gonorrhoeae/genetics , Neisseria gonorrhoeae/pathogenicity , Reproductive Tract Infections/immunology , Toll-Like Receptor 4 , Vagina/metabolism
15.
Carbohydr Polym ; 99: 692-9, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24274560

ABSTRACT

Rapid initiation of clotting is critical to trauma patients. In the present study exopolymers (EPs) from four desert cyanobacteria including Tolypothrix tenuis and three species of Anabaena have been discovered as potential hemostatic biomaterials. The EPs showed reduction in activated partial thromboplastin time (APTT) and prothrombin time (PT) by 16-41% and 12-65%, respectively. Besides hastening blood clotting, the EPs could absorb 7.1-25.9 g H2O g(-1) EP and displayed 7.1-18.1% hydrophobicity. They were noncytotoxic and biodegradable. The EP from Anabaena sp. showed strong antibacterial activity against E. coli, S. aureus and B. licheniformis. These results suggest that cyanobacteria, the microscopic phototrophs growing rapidly over simple mineral medium could prove to be a novel source of affordable hemostatic dressings for the traumatic wounds in underdeveloped and developing countries. Compositional analysis of the EPs showed them to be consisting of mainly carbohydrate (17-50%), protein (4.4-7.2%), uronic acid (4.7-9.5%) and sulphate (0.6-6.6%). Their viscometric molecular weight ranged from 539 to 3679 kDa. They were further characterized using GC-MS and FTIR.


Subject(s)
Anabaena/chemistry , Anti-Bacterial Agents/pharmacology , Biocompatible Materials/pharmacology , Blood Coagulation/drug effects , Cyanobacteria/chemistry , Hemostatics/pharmacology , Polymers/pharmacology , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/isolation & purification , Bacillus/drug effects , Bacillus/growth & development , Biocompatible Materials/chemistry , Biocompatible Materials/isolation & purification , Escherichia coli/drug effects , Escherichia coli/growth & development , Gas Chromatography-Mass Spectrometry , Hemostatics/chemistry , Hemostatics/isolation & purification , Humans , Partial Thromboplastin Time , Polymers/chemistry , Polymers/isolation & purification , Prothrombin Time , Spectroscopy, Fourier Transform Infrared , Staphylococcus aureus/drug effects , Staphylococcus aureus/growth & development
16.
mBio ; 4(6): e00892-13, 2013 Nov 19.
Article in English | MEDLINE | ID: mdl-24255126

ABSTRACT

UNLABELLED: Phosphoethanolamine (PEA) on Neisseria gonorrhoeae lipid A influences gonococcal inflammatory signaling and susceptibility to innate host defenses in in vitro models. Here, we evaluated the role of PEA-decorated gonococcal lipid A in competitive infections in female mice and in male volunteers. We inoculated mice and men with mixtures of wild-type N. gonorrhoeae and an isogenic mutant that lacks the PEA transferase, LptA. LptA production conferred a marked survival advantage for wild-type gonococci in the murine female genital tract and in the human male urethra. Our studies translate results from test tube to animal model and into the human host and demonstrate the utility of the mouse model for studies of virulence factors of the human-specific pathogen N. gonorrhoeae that interact with non-host-restricted elements of innate immunity. These results validate the use of gonococcal LptA as a potential target for development of novel immunoprophylactic strategies or antimicrobial treatments. IMPORTANCE: Gonorrhea is one of the most common bacterial sexually transmitted infections, and increasing antibiotic resistance threatens the use of currently available antimicrobial therapies. In this work, encompassing in vitro studies and in vivo studies of animal and human models of experimental genital tract infection, we document the importance of lipid A's structure, mediated by a single bacterial enzyme, LptA, in enhancing the fitness of Neisseria gonorrhoeae. The results of these studies suggest that novel agents targeting LptA may offer urgently needed prevention or treatment strategies for gonorrhea.


Subject(s)
Ethanolamines/analysis , Gonorrhea/microbiology , Lipid A/chemistry , Lipid A/metabolism , Neisseria gonorrhoeae/physiology , Animals , Disease Models, Animal , Ethanolaminephosphotransferase/genetics , Ethanolaminephosphotransferase/metabolism , Female , Gene Knockout Techniques , Healthy Volunteers , Humans , Male , Mice , Microbial Viability , Neisseria gonorrhoeae/chemistry , Neisseria gonorrhoeae/enzymology , Neisseria gonorrhoeae/pathogenicity , Virulence , Virulence Factors/genetics , Virulence Factors/metabolism
17.
J Mol Biol ; 425(18): 3389-402, 2013 Sep 23.
Article in English | MEDLINE | ID: mdl-23810904

ABSTRACT

Gram-negative bacteria possess an outer membrane envelope consisting of an outer leaflet of lipopolysaccharides, also called endotoxins, which protect the pathogen from antimicrobial peptides and have multifaceted roles in virulence. Lipopolysaccharide consists of a glycan moiety attached to lipid A, embedded in the outer membrane. Modification of the lipid A headgroups by phosphoethanolamine (PEA) or 4-amino-arabinose residues increases resistance to the cationic cyclic polypeptide antibiotic, polymyxin. Lipid A PEA transferases are members of the YhjW/YjdB/YijP superfamily and usually consist of a transmembrane domain anchoring the enzyme to the periplasmic face of the cytoplasmic membrane attached to a soluble catalytic domain. The crystal structure of the soluble domain of the protein of the lipid A PEA transferase from Neisseria meningitidis has been determined crystallographically and refined to 1.4Å resolution. The structure reveals a core hydrolase fold similar to that of alkaline phosphatase. Loop regions in the structure differ, presumably to enable interaction with the membrane-localized substrates and to provide substrate specificity. A phosphorylated form of the putative nucleophile, Thr280, is observed. Metal ions present in the active site are coordinated to Thr280 and to residues conserved among the family of transferases. The structure reveals the protein components needed for the transferase chemistry; however, substrate-binding regions are not evident and are likely to reside in the transmembrane domain of the protein.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Ethanolaminephosphotransferase/chemistry , Neisseria meningitidis/enzymology , Polymyxins/pharmacology , Binding Sites , Ethanolaminephosphotransferase/genetics , Ethanolaminephosphotransferase/metabolism , Ethanolamines/metabolism , Lipid A/metabolism , Lipopolysaccharides/metabolism , Models, Biological , Models, Molecular , Neisseria meningitidis/drug effects , Neisseria meningitidis/genetics , Protein Interaction Domains and Motifs/genetics , Protein Structure, Quaternary , Protein Structure, Secondary/physiology
18.
Glycobiology ; 23(7): 820-32, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23493680

ABSTRACT

Endolysins are bacteriophage enzymes that lyse their bacterial host for phage progeny release. They commonly contain an N-terminal catalytic domain that hydrolyzes bacterial peptidoglycan (PG) and a C-terminal cell wall-binding domain (CBD) that confers enzyme localization to the PG substrate. Two endolysins, phage lysin L (PlyL) and phage lysin G (PlyG), are specific for Bacillus anthracis. To date, the cell wall ligands for their C-terminal CBD have not been identified. We recently described structures for a number of secondary cell wall polysaccharides (SCWPs) from B. anthracis and B. cereus strains. They are covalently bound to the PG and are comprised of a -ManNAc-GlcNAc-HexNAc- backbone with various galactosyl or glucosyl substitutions. Surface plasmon resonance (SPR) showed that the endolysins PlyL and PlyG bind to the SCWP from B. anthracis (SCWPBa) with high affinity (i.e. in the µM range with dissociation constants ranging from 0.81 × 10(-6) to 7.51 × 10(-6) M). In addition, the PlyL and PlyG SCWPBa binding sites reside with their C-terminal domains. The dissociation constants for the interactions of these endolysins and their derived C-terminal domains with the SCWPBa were in the range reported for other protein-carbohydrate interactions. Our findings show that the SCWPBa is the ligand that confers PlyL and PlyG lysin binding and localization to the PG. PlyL and PlyG also bound the SCWP from B. cereus G9241 with comparable affinities to SCWPBa. No detectable binding was found to the SCWPs from B. cereus ATCC (American Type Culture Collection) 10987 and ATCC 14579, thus demonstrating specificity of lysin binding to SCWPs.


Subject(s)
Amidohydrolases/metabolism , Bacillus anthracis/metabolism , Bacterial Proteins/metabolism , Cell Wall/chemistry , N-Acetylmuramoyl-L-alanine Amidase/metabolism , Polysaccharides, Bacterial/metabolism , Viral Proteins/metabolism , Amidohydrolases/chemistry , Amino Sugars/chemistry , Bacillus anthracis/chemistry , Bacterial Proteins/chemistry , Binding Sites , Cell Wall/metabolism , Hexoses/chemistry , Ligands , N-Acetylmuramoyl-L-alanine Amidase/chemistry , Polysaccharides, Bacterial/chemistry , Protein Binding , Protein Structure, Tertiary , Viral Proteins/chemistry
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