Use Of Bacterial Cells Research Articles

Bacterial Synthesis of Ternary CdSAg Quantum Dots through Cation Exchange: Tuning the Composition and Properties of Biological Nanoparticles for Bioimaging and Photovoltaic Applications.

In this study, we introduce a biological method for the production of ternary Quantum Dots (QDs): complex nanostructures with tunable optical and structural properties that utilizes post-synthesis modifications through cation exchange. This versatile in-situ cation exchange method being reported for the first time shows great potential for extending the scope of microbial synthesis. By using this bacterial-based method, we easily synthesize and purify CdS, CdSAg, and Ag2S nanocrystals of a size below 15 nm and with variable morphologies that exhibit fluorescence emissions covering a broad spectral range (from 400 to 800 nm). Energy-dispersive X-ray spectroscopy (EDS) results indicate the partial replacement of Cd2+ by Ag+ when AgNO3 concentration is increased. This replacement produces CdSAg ternary QDs hetero-structures with high stability, fluorescence in the NIR-I (700 - 800 nm), and 36.13% quantum yield. Furthermore, this reaction can be extended for the production of soluble Ag2S nanoparticles (NPs) without any traces of Cd. QDs biosynthesized through this cation exchange process display very low toxicity when tested in bacterial or human cell lines. Biosynthesized ternary hetero-structures were used as red fluorescent dyes to label HeLa cells in confocal microscopy studies, which validates its use in bioimaging applications in the near infrared region. In addition, the application of biologically-produced cadmium NPs in solar cells is reported for the first time. The three biosynthesized QDs were successfully used as photosensitizers, where the CdSAg QDs show the best photovoltaic parameters. Altogether, obtained results validate the use of bacterial cells for the controlled production of nanomaterials with properties that allow their application in diverse technologies. We developed a simple biological process for obtaining tunable Quantum Dots (QDs) with different metal compositions through a cation exchange process. Nanoparticles (NPs) are produced in the extracellular space of bacterial cells exposed to cysteine and CdCl2 in a reaction that depends on S2− generation mediated by cysteine desulfhydrase enzymes and uses cellular biomolecules to stabilize the nanoparticle. Using this extracellular approach, water-soluble fluorescent CdS, CdSAg, and Ag2S Quantum Dots with a tunable emission ranging from 400 to 800 nm were generated. This is the first study reporting the use of microorganisms to produce tunable ternary QDs and the first time that a cation exchange process mediated by cells is described. Obtained results validate the use of biological synthesis to produce NPs with new characteristics and opens a completely new research field related to the use of microorganisms to synthesize complex NPs that are difficult to obtain with regular chemical methods.

Biological Synthesis of CdS/CdSe Core/Shell Nanoparticles and Its Application in Quantum Dot Sensitized Solar Cells.

In the present work, we report the use of bacterial cells for the production of CdS/CdSe Core/Shell quantum dots (QDs), a complex nanostructure specially designed to improve their performance as photosensitizer in photovoltaic devices. The method requires the incorporation of L-cysteine, CdCl2 and Na2SeO3 to Escherichia coli cultures and allows a tight control of QDs properties. The obtained CdS/CdSe QDs were photophysically and structurally characterized. When compared to CdS QDs, the classical shift in the UV-visible spectra of Core/Shell nanostructures was observed in CdS/CdSe QDs. The nanosize, structure, and composition of Core/Shell QDs were confirmed by TEM and EDS analysis. QDs presented a size of approximately 12 nm (CdS) and 17 nm (CdS/CdSe) as determined by dynamic light scattering (DLS), whereas the fourier transform infrared (FTIR) spectra allowed to distinguish the presence of different biomolecules bound to both types of nanoparticles. An increased photostability was observed in CdS/CdSe nanoparticles when compared to CdS QDs. Finally, biosynthesized CdS/CdSe Core/Shell QDs were used as photosensitizers for quantum dots sensitized solar cells (QDSSCs) and their photovoltaic parameters determined. As expected, the efficiency of solar cells sensitized with biological CdS/CdSe QDs increased almost 2.5 times when compared to cells sensitized with CdS QDs. This work is the first report of biological synthesis of CdS/CdSe Core/Shell QDs using bacterial cells and represents a significant contribution to the development of green and low-cost photovoltaic technologies.

Metal ion binding and tolerance of bacteria cells in view of sensor applications

Abstract. The biotechnological use of bacterial cells and cell components for the detection and accumulation of valuable substances, such as metals and rare earth elements in aqueous systems, is possible by utilizing innate binding characteristics of microorganisms. We have studied the bacteria cells of Lysinibacillus sphaericus JG-B53 and Sporosarcina ureae ATCC 13881 to assess their potential applicability for the detection of rare earth elements, base metals or precious metals in water. First, we have demonstrated the interactions of the cells with the metal complexes of Au, Ho and Y by studying the color changes of the respective solutions, scanning electron microscopy (SEM) imaging of the metal cluster decoration on the cell surfaces and cell growth tolerance tests. Based on these results, we have developed two potential sensor systems. A colorimetric sensor was established by applying gold nanoparticles (AuNPs) functionalized with surface-layer (S-layer) proteins SslA of S. ureae ATCC 13881 or Slp1 of L. sphaericus JG-B53 for the selective detection of YCl3 up to 1.67 × 10−5 mol L−1 in water. Additionally, a regenerative sensor layer of S-layer proteins on a thin gold film was developed for the detection of 1 × 10−4 mol L−1 YCl3 in water by surface plasmon resonance (SPR) spectroscopy.

Unleashing the potential of ligninolytic bacterial contributions towards pulp and paper industry: key challenges and new insights.

Lignocellulose biomass predominantly constitutes the main feedstock for pulp and paper industry. Though some products of pulp and paper industry require the presence of lignin content, for most of the useful products formation lies in the efficient and selective removal of lignin component to make use of the intact cellulose fraction during the pretreatment of pulp. Lignin is a recalcitrant heteropolymer comprised of several complex stable bonds and linkages. The chemicals or intense energy processes used for delignification process release the hazardous chemicals compounds in the wastewater which cause toxicity and environmental pollution. The implementation of bacterial species has elucidated an effective approach in the generation of value-added products while degrading lignin from pulp biomass as well as detoxification of effluent. The direct use of bacterial cells in lignocellulose biomass and wastewater streams is promising as it outperforms the practical and technical constraints largely confronted by fungal and enzymatic means. The present review paper thus unleashed the potential of ligninolytic bacteria towards delignification of pulp biomass and treatment of effluent together with bioconversion of biomass and lignin into value-added products. Graphical abstract Schematic illustration of potential possible contribution of ligninolytic bacteria towards pulp and paper industry.

"Use of acidophilic bacteria of the genus Acidithiobacillus to biosynthesize CdS fluorescent nanoparticles (quantum dots) with high tolerance to acidic pH".

The use of bacterial cells to produce fluorescent semiconductor nanoparticles (quantum dots, QDs) represents a green alternative with promising economic potential. In the present work, we report for the first time the biosynthesis of CdS QDs by acidophilic bacteria of the Acidithiobacillus genus. CdS QDs were obtained by exposing A. ferrooxidans, A. thiooxidans and A. caldus cells to sublethal Cd2+ concentrations in the presence of cysteine and glutathione. The fluorescence of cadmium-exposed cells moves from green to red with incubation time, a characteristic property of QDs associated with nanocrystals growth. Biosynthesized nanoparticles (NPs) display an absorption peak at 360nm and a broad emission spectra between 450 and 650nm when excited at 370nm, both characteristic of CdS QDs. Average sizes of 6 and 10nm were determined for green and red NPs, respectively. The importance of cysteine and glutathione on QDs biosynthesis in Acidithiobacillus was related with the generation of H2S. Interestingly, QDs produced by acidophilic bacteria display high tolerance to acidic pH. Absorbance and fluorescence properties of QDs was not affected at pH 2.0, a condition that totally inhibits the fluorescence of QDs produced chemically or biosynthesized by mesophilic bacteria (stable until pH 4.5-5.0). Results presented here constitute the first report of the generation of QDs with improved properties by using extremophile microorganisms.

Oral Administration of Lactococcus lactis Expressing Synthetic Genes of Myelin Antigens in Decreasing Experimental Autoimmune Encephalomyelitis in Rats.

BackgroundMultiple sclerosis is a human autoimmunological disease that causes neurodegeneration. One of the potential ways to stop its development is induction of oral tolerance, whose effect lies in decreasing immune response to the fed antigen. It was shown in animal models that administration of specific epitopes of the three main myelin proteins – myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP), and proteolipid protein (PLP) – results in induction of oral tolerance and suppression of disease symptoms. Use of bacterial cells to produce and deliver antigens to gut mucosa seems to be an attractive method for oral tolerance induction in treatment of diseases with autoimmune background.Material/MethodsSynthetic genes of MOG35-55, MBP85-97, and PLP139-151 myelin epitopes were generated and cloned in Lactococcus lactis under a CcpA-regulated promoter. The tolerogenic effect of bacterial preparations was tested on experimental autoimmune encephalomyelitis, which is the animal model of MS. EAE was induced in rats by intradermal injection of guinea pig spinal cord homogenate into hind paws.ResultsRats were administered preparations containing whole-cell lysates of L. lactis producing myelin antigens using different feeding schemes. Our study demonstrates that 20-fold, but not 4-fold, intragastric administration of autoantigen-expressing L. lactis cells under specific conditions reduces the clinical symptoms of EAE in rats.ConclusionsThe present study evaluated the use of myelin antigens produced in L. lactis in inhibiting the onset of experimental autoimmune encephalomyelitis in rats. Obtained results indicate that application of such recombinant cells can be an attractive method of oral tolerance induction.

Gold(III) Reduction by the Rhizobacterium Azospirillum brasilense with the Formation of Gold Nanoparticles

For the soil nitrogen-fixing bacterium Azospirillum brasilense, the ability to reduce [AuCl4](-) and to form gold nanoparticles (GNPs) has been demonstrated, with the appearance of a mauve tint of the culture. To validate the shapes and chemical nature of nanoparticles, transmission electron microscopy (TEM) and X-ray fluorescence analysis were used. For the widely studied agriculturally important wild-type strains A. brasilense Sp7 and Sp245, GNPs formed after 10days of incubation of cell biomass with 0.25mM [AuCl4](-) were shown (using TEM) to be mainly of spherical form (5 to 20nm in diameter), with rare occasional triangles. In the course of cultivation with [AuCl4](-), after 5days, a mauve tint was already visible for cells of strain Sp245.5, after 6days for Sp245 and after 10days for Sp7. Thus, for the mutant strain Sp245.5 (which has significant differences in the structure and composition of cell-surface polysaccharides as compared with Sp245), a more rapid formation of GNPs was observed. Moreover, their TEM images (also obtained after 10days) showed different shapes: nano-sized spheres, triangles, hexagons and rods, as well as larger round-shaped flower-like nanoparticles about 100nm in size. Since by the time of GNP formation in our experiments the cells were found to be already not viable, this confirms the dominating role of cell surface structure and chemical composition in shaping the GNPs formed in the course of [AuCl4](-) reduction to Au(0). This finding may be useful for understanding the natural biogeochemical mechanisms of gold reduction and formation of GNPs involving microorganisms. The data obtained may also help in developing protocols for environmentally friendly synthesis of nanoparticles and possible use of bacterial cells with modified surface structure and composition for their fabrication.

Enhancing Jatropha oil extraction yield from the kernels assisted by a xylan-degrading bacterium to preserve protein structure.

We investigated the use of bacterial cells isolated from paddy crab for the extraction of oil from Jatropha seed kernels in aqueous media while simultaneously preserving the protein structures of this protein-rich endosperm. A bacterial strain—which was marked as MB4 and identified by means of 16S rDNA sequencing and physiological characterization as either Bacillus pumilus or Bacillus altitudinis—enhanced the extraction yield of Jatropha oil. The incubation of an MB4 starter culture with preheated kernel slurry in aqueous media with the initial pH of 5.5 at 37 °C for 6 h liberated 73% w/w of the Jatropha oil. Since MB4 produces xylanases, it is suggested that strain MB4 facilitates oil liberation via degradation of hemicelluloses which form the oil-containing cell wall structure of the kernel. After MB4 assisted oil extraction, SDS-PAGE analysis showed that the majority of Jatropha proteins were preserved in the solid phase of the extraction residues. The advantages offered by this process are: protein in the residue can be further processed for other applications, no purified enzyme preparation is needed, and the resulting oil can be used for biodiesel production.

Improved strength and durability of fly ash-amended concrete by microbial calcite precipitation

Fly ash acts as a partial replacement material for both Portland cement and fine aggregate. An innovative approach of microbial calcite precipitation in fly ash-amended concrete has been investigated. This is the first report to discuss the role of microbial calcite precipitation in enhancing the durability of fly ash-amended concrete. The present study investigated the effects of Bacillus megaterium ATCC 14581 on compressive strength, water absorption and water impermeability of fly ash-amended mortar and concrete. Mortar specimens were used for compressive strength and water absorption tests, while concrete specimens were used for water impermeability tests. At the fly ash concentrations of 10%, 20% and 40% in mortars, bacterial cell enhanced mortar compressive strength by 19%, 14% and 10%, respectively, compared to control specimens. Treated mortar cubes absorbed more than three times less water than control cubes as a result of microbial calcite deposition. Microbial deposition of a layer of calcite on the surface of the concrete specimens resulted in substantial decrease of water uptake and permeability compared to control specimens without bacteria. Microbial cells also prevented ingress of water effectively in different concentrations of fly ash-amended concrete. Scanning Electron Micrography (SEM) analyses evidenced the direct involvement of bacteria in calcite precipitation. The approach of the present study gives us dual environment friendly advantages. First, use of fly ash-a recovered resource reduces depletion of natural resources and also reduces the energy-intensive manufacturing of other concrete ingredients, leading to savings in both energy usage and emissions of greenhouse gases. And second, use of bacterial cells to improve strength and durability of fly ash-amended concrete further provides greener and economic options.

Live bacterial cells as analytical tools for speciation analysis: Hypothetical or practical?

This review tries to show the capability of live bacterial cells in speciation analysis, so it emphasizes speciation analysis of metalloid and organometalloid species by combining an uptake process by live bacterial cells with a specific detector. Especially covered in depth is the uptake process, because better understanding of equilibrium and retention mechanisms provides the basis for optimizing the use of bacterial cells. We outline the main parameters affecting the uptake process and the analytical characteristics. We also cover bacteria-based biosensors that represent a research field of growing interest in using live bacteria. We also highlight some future considerations about the role of recombinant bacterial cells in analytical chemistry.

Bacterial cell microarrays for the detection and characterization of antibodies against surface antigens

Bacterial cell surface antigens interact with the host immune system resulting in the production of antibodies. Detection of antibodies against surface antigens has applications in diagnosis of many bacterial infections, assessment of immune status and epidemiological studies. We developed a microarray platform, for antibody detection, by printing Gram-negative and Gram-positive whole bacterial cells on nitrocellulose coated glass substrates. Antibody binding was detected using fluorophore labeled secondary antibodies. The sensitivity of antibody detection was found to be 0.1 μg/ml. Using bacterial cell microarrays it was also possible to successfully detect antibodies against Francisella tularensis in canine serum samples declared positive for tularemia based on microagglutination antibody titer. Use of bacterial cells as the antigen source in immunoassays has the advantages of simulating in vivo presentation of surface antigens and also eliminating the need for antigen purification. The microarray format gives the added advantage of simultaneous detection of antibodies against multiple bacteria employing only small amounts of samples and reagents.

Artificial Cell Therapy: New Strategies for the Therapeutic Delivery of Live Bacteria.

There has been rapid growth in research regarding the use of live bacterial cells for therapeutic purposes. The recognition that these cells can be genetically engineered to synthesize products that have therapeutic potential has generated considerable interest and excitement among clinicians and health professionals. It is expected that a wide range of disease modifying substrates such as enzymes, hormones, antibodies, vaccines, and other genetic products will be used successfully and will impact upon health care substantially. However, a major limitation in the use of these bacterial cells is the complexity of delivering them to the correct target tissues. Oral delivery of live cells, lyophilized cells, and immobilized cells has been attempted but with limited success. Primarily, this is because bacterial cells are incapable of surviving passage through the gastrointestinal tract. In many occasions, when given orally, these cells have been found to provoke immunogenic responses that are undesirable. Recent studies show that these problems can be overcome by delivering live bacterial cells, such as genetically engineered cells, using artificial cell microcapsules. This review summarizes recent advances in the therapeutic use of live bacterial cells for therapy, discusses the principles of using artificial cells for the oral delivery of bacterial cells, outlines methods for preparing suitable artificial cells for this purpose, addresses potentials and limitations for their application in therapy, and provides insight for the future direction of this emergent and highly prospective technology.

Fractionation of rat IgG subclasses and screening for IgG Fc-binding to bacteria

The four IgG subclasses of the rat, IgG1, IgG2a, IgG2b and IgG2c, were purified from normal serum by a combination of protein A-affinity chromatography and DEAE-cellulose chromatography. Purified, radiolabelled preparations of IgG were tested for binding to Gram-positive bacteria representing five different Fc-receptor (FcR) types. Distinct rat subclass-specific Fc-binding was noted to bacterial species belonging to different Fc-receptor types. Staphylococcus aureus (FcR I) strains bind IgGl and IgG2c as shown by others. Group C and G Streptococci (FcR III) bind all four subclasses of rat IgG. Streptococcus zooepidemicus strains (FcR V) also bind all four subclasses but only to a lower degree. Human group A Streptococci (FcR II) and bovine group G Streptococci (FcR IV) do not bind any of the rat IgG subclasses. Elution studies on two strains, Staphylococcus aureus, Cowan I, and human group G Streptococcus, G 148, showed that both thiocyanate and pH-elution might be useful for the fractionation of IgG subclasses bound to bacterial cells. The present work indicates the possible use of bacterial cells as solid-phase absorbents in immunological studies of rat IgG.