Biosynthesis, purification and characterization of silver nanoparticles using Escherichia coli

The application of nanoscale materials and structures, usually ranging from 1 to 100 nanometers (nm), is an emerging area of nanoscience and nanotechnology. Nanomaterials may provide solutions to technological and environmental challenges in the areas of solar energy conversion, catalysis, medicine, and water-treatment. The development of techniques for the controlled synthesis of nanoparticles of well-defined size, shape and composition, to be used in the biomedical field and areas such as optics and electronics, has become a big challenge. Development of reliable and eco-friendly processes for synthesis of metallic nanoparticles is an important step in the field of application of nanotechnology. One of the options to achieve this objective is to use ‘natural factories’ such as biological systems. This study reports the optimal conditions for maximum synthesis of silver nanoparticles (AgNPs) through reduction of Ag⁺ ions by the culture supernatant of Escherichia coli. The synthesized silver nanoparticles were purified by using sucrose density gradient centrifugation. The purified sample was further characterized by UV–vis spectra, fluorescence spectroscopy and TEM. The purified solution yielded the maximum absorbance peak at 420 nm and the TEM characterization showed a uniform distribution of nanoparticles, with an average size of 50 nm. X-ray diffraction (XRD) spectrum of the silver nanoparticles exhibited 2θ values corresponding to the silver nanocrystal. The size-distribution of nanoparticles was determined using a particle-size analyzer and the average particle size was found to be 50 nm. This study also demonstrates that particle size could be controlled by varying the parameters such as temperature, pH and concentration of AgNO₃.     

Bacteria and bacteriophage inactivation by silver and zinc oxide nanoparticles

Microbial contaminants such as bacteria and viruses are of great concern in water. As nanotechnology continues to grow, understanding the interactions of nanoparticles with bacteria and viruses is important to protect public health and the environment. In this study, the effect of two commonly used nanoparticles, silver nanoparticles (AgNPs, average particle size=21 nm) and zinc oxide nanoparticles (ZnO NPs, average particle size=39 nm), on the growth of bacteria (Eschericia coli) and bacteriophages (MS2) were evaluated using a standard double agar layer (DAL) method and a turbidimetric microtiter assay. A 1-h prior exposure of MS2 to nanoparticles did not inactivate MS2 at the highest nanoparticle concentrations tested (5mg/L total Ag and 20 mg/L ZnO). No bacteriophage inactivation was observed in the presence of AgNPs, Ag(+)/AgNPs (50:50 in mass ratio) or Ag(+) ions, all at the total Ag concentration of 5mg/L. In a binary (bacteria-phages) system where the E. coli host was exposed to MS2 and nanoparticles simultaneously, the dynamic changes of active bacteria and MS2 phages during incubation demonstrated that exposure of AgNPs (5mg/L Ag) and ZnO NPs (20mg/L ZnO) increased the number of phages by 2-6 orders of magnitude. These results suggested that exposure of nanoparticles could greatly facilitate bacterial viruses like MS2 to infect the E. coli host.     

Green synthesis of silver nanoparticles using seed extract of Jatropha curcas

An eco-friendly process for rapid synthesis of silver nanoparticles has been reported using aqueous seed extract of Jatropha curcas. Formation of stable silver nanoparticles at different concentration of AgNO₃ gives mostly spherical particles with diameter ranging from 15 to 50 nm. The resulting silver particles are characterized using HRTEM, XRD and UV–vis spectroscopic techniques. XRD study shows that the particles are crystalline in nature with face centered cubic geometry.     

Comparative evaluation of antibacterial activity of silver nanoparticles synthesized using Rhizophora apiculata and glucose

The focus of the study is to compare the antibacterial efficacy of silver nanoparticles (AgNPs) fabricated by exploiting biological (a mangrove plant, Rhizophora apiculata) and chemical means (Glucose). The synthesized nanoparticles were characterised using UV-visible absorption spectrophotometry (UV-vis), Fourier transform Infra-red Spectroscopy (FTIR) and Transmission electron microscopy (TEM). Biologically synthesized silver nanoparticles (BAgNPs) were observed at 423 nm with particle sizes of 19-42 nm. The chemically synthesized silver nanoparticles (CAgNPs) showed a maximum peak at 422 nm with particle sizes of 13-19 nm. An obvious superiority of the antibacterial potency of BAgNPs compared to the CAgNPs as denoted by the zone of inhibition (ZoI) was noted when the nanoparticles were treated against seven different Microbial Type Culture Collection (MTCC) strains. The current study therefore elucidates that the synthesized AgNPs were efficient against the bacterial strains tested.     

中空碳球负载TiO2纳米颗粒用于增强光催化抗菌性能

以聚吡咯包覆聚苯乙烯核壳结构衍生的中空碳球(hollow carbon spheres,HCS)为载体,乙酰丙酮钛(TOAC)为钛源,通过湿化学法和可控热解法制备了HCS负载纳米二氧化钛(HCS@TiO₂)复合材料。通过粉末X射线衍射、紫外可见光谱、X射线光电子能谱、热重分析、扫描电子显微镜、透射电子显微镜、光致发光光谱和Mott-Schottky曲线对HCS@TiO₂的晶态结构、微观形貌、光学性能等进行表征,并通过调控TiO₂的负载量和热解温度对HCS@TiO₂复合材料进行了结构优化。在模拟太阳光照射条件下,以大肠杆菌(E. coli)和金黄色葡萄球菌(S. aureus)为实验对象,研究了不同TiO₂负载量的HCS@TiO₂复合材料及TiO₂和HCS对照样品的光催化抗菌性能,以及光照时间对抗菌性能的影响。结果表明,当TOAC与HCS质量比为15∶1、热解温度为650℃时,最优化的HCS@TiO₂-15复合材料...     

Plant extract mediated synthesis of silver and gold nanoparticles and its antibacterial activity against clinically isolated pathogens

Biosynthesis of nanoparticles is under exploration is due to wide biomedical applications and research interest in nanotechnology. Bioreduction of silver nitrate (AgNO(3)) and chloroauric acid (HAuCl(4)) for the synthesis of silver and gold nanoparticles respectively with the plant extract, Mentha piperita (Lamiaceae). The plant extract is mixed with AgNO(3) and HAuCl(2), incubated and studied synthesis of nanoparticles using UV-Vis spectroscopy. The nanoparticles were characterized by FTIR, SEM equipped with EDS. The silver nanoparticles synthesized were generally found to be spherical in shape with 90 nm, whereas the synthesized gold nanoparticles were found to be 150 nm. The results showed that the leaf extract of menthol is very good bioreductant for the synthesis of silver and gold nanoparticles and synthesized nanoparticles active against clinically isolated human pathogens, Staphylococcus aureus and Escherichia coli.     

Schiff base stabilized silver nanoparticles as potential sensor for Hg(II) detection,and anticancer and antibacterial agent

This study reports a facile synthesis of silver nanoparticles (C3-AgNPs) by chemical route, using C3; 2,2′-((1E,1′E)-(propane-1,3-diylbis(azanylylidene))bis(methanylylidene))diphenol (3) and silver nitrate. The formation of nanoparticles was monitored using UV–Vis spectroscopy by the appearance of typical surface plasmon absorption maxima. The synthesized C3-AgNPs were characterized using Fourier-Transform-infrared (FTIR) and atomic force microscopy (AFM) techniques. In addition, the effect of concentration, temperature, time, pH, and stability in salts solution on C3-AgNPs was determined. From AFM, C3-AgNPs were found polydispersed with average size of 29.93 nm. Furthermore, the study reports C3-AgNPs as sensitive protocol for the detection of toxic metal; Hg(II) in tap water. From ten salts tested, C3-AgNPs demonstrated a sensitive and selective spectrophotometric signal and aggregation induced decrease of surface plasmon resonance (SPR) band. The nanosensor probe displayed a sensitive response to Hg(II) in a wide range of concentrations and pH. In addition, the decrease in SPR band of C3-AgNPs due to Hg(II) was not affected by tap water samples. C3-AgNPs also exhibited a redox catalytic potential in dyes degradation. In biological application, C3-AgNPs exhibited significant anticancer and antibacterial potential of 65 to 94% at 24–72 h, and inhibition zone of 7–18 mm, respectively. Hence, the synthesized C3-AgNPs could have promising application in environmental and pharmacological remediation.     

Combining biofunctional magnetic nanoparticles and ATP bioluminescence for rapid detection of Escherichia coli

A rapid, specific and sensitive method for assay of Escherichia coli (E. coli) using biofunctional magnetic nanoparticles (BMNPs) in combination with adenosine triphosphate (ATP) bioluminescence was proposed. The BMNPs were fabricated by immobilizing a specific anti-E. coli antibody on the surface of amine-functionalized magnetic nanoparticles (about 20 nm in diameter), and then was applied to capture the target bacteria E. coli from samples. The BMNPs exhibited high capture efficiency to E. coli. Transmission electron microscope (TEM) images showed that the BMNPs were bound to the surface of entire E. coli cells. The target bacteria became magnetic so that could be isolated easily from the sample solution by employing an external magnetic field. The concentration of E. coli captured by the BMNPs was then detected by an ATP bioluminescence method. The optimization of ATP measurement was carried out to improve the detection sensitivity. The proposed method was applied to detect the E. coli inoculated into pasteurized milk with low detection limit (20 cfu/mL) and short detection time (about 1 h).     

A conductometric immunosensor based on functionalized magnetite nanoparticles for E. coli detection

We report a new approach for immunoassays based on magnetite nanoparticles for Escherichia coli (E. coli) detection using conductometric measurements. Biotinylated antibodies, anti-E. coli, were immobilized on streptavidin modified magnetite nanoparticles by biotin–streptavidin interaction. A layer of functionalized nanoparticles were directly immobilized on the conductometric electrode using glutaraldehyde cross-linking.The specific test with E. coli cells and the non specific test using Staphylococcus epidermidis (S. epidermidis) were investigated by conductometric measurements. Results show a good response as a function of antigen additions. The detection of 1 CFU/ml of E. coli induces a conductivity variation of 35 μS. The negative test shows good selectivity using the conductometric immunosensor. Conductometric measurements allow to detect 500 CFU/l.     

Biomimetic preparation of polymer-supported free radical scavenging, cytocompatible and antimicrobial "green" silver nanoparticles using aqueous extract of Citrus sinensis peel

In the pursuit of making the nanoscale-research greener, the utilization of the reductive potency of a common byproduct of food processing industry i.e. orange peel is reported here to prepare biopolymer-templated "green" silver nanoparticles. Aqueous extract of orange peel at basic pH was exploited to prepare starch supported nanoparticles under ambient conditions. The compositional abundance of pectins, flavonoids, ascorbic acid, sugars, carotenoids and myriad other flavones may be envisaged for the effective reductive potential of orange peel to generate silver nanoparticles. The nanoparticles were distributed within a narrow size spectrum of (3-12 nm) with characteristic Bragg's reflection planes of fcc structure, and surface plasmon resonance peak at 404 nm. Anti-lipid peroxidation assay using goat liver homogenate and DPPH scavenging test established the anti-oxidant potency of the silver nanoparticles. Their synergy with rifampicin against Bacillus subtilis MTCC 736 and cytocompatibility with the human leukemic monocytic cell line, THP-1 were also investigated. Thus, the present work deals with the preparation of starch assisted anti-microbial, cytocompatible and free radical scavenging "green" silver nanoparticles.     

Electrophoretic deposition as a tool for separation of protein inclusion bodies from host bacteria in suspension

This paper shows, for the first time, that the electrophoretic deposition technique is able to selectively collect protein inclusion bodies (PBs) from the host bacteria suspensions. In the first step, zeta potential as a function of pH is carefully determined for both species involved. Based on the obtained dependencies, the pH of the mixture of PBs and bacteria is precisely adjusted and the electrophoretic experiment is carried out. We show that the efficiency of separation and the yield depends not only on the electrokinetic properties of given species but also on the electrode composition and surface morphology. The deposited species are easily removed by forced washing or reverse electric field. As a whole, the selectivity and the yields are higher than in most alternative state-of-the art techniques.     

Cinnamon zeylanicum bark extract and powder mediated green synthesis of nano-crystalline silver particles and its bactericidal activity

The exploitation of various plant materials for the biosynthesis of nanoparticles is considered a green technology as it does not involve any harmful chemicals. The present study reports the synthesis of silver (Ag) nanoparticles from silver precursor using the bark extract and powder of novel Cinnamon zeylanicum. Water-soluble organics present in the plant materials were mainly responsible for the reduction of silver ions to nano-sized Ag particles. TEM and XRD results confirmed the presence of nano-crystalline Ag particles. The pH played a major role in size control of the particles. Bark extract produced more Ag nanoparticles than the powder did, which was attributed to the large availability of the reducing agents in the extract. Zeta potential studies showed that the surface charge of the formed nanoparticles was highly negative. The EC₅₀ value of the synthesized nanoparticles against Escherichia coli BL-21 strain was 11 ± 1.72 mg/L. ThusC. zeylanicum bark extract and powder are a good bio-resource/biomaterial for the synthesis of Ag nanoparticles with antimicrobial activity.     

Phytosynthesis of Au, Ag and Au-Ag bimetallic nanoparticles using aqueous extract and dried leaf of Anacardium occidentale

Present study reports a green chemistry approach for the biosynthesis of Au, Ag, Au-Ag alloy and Au core-Ag shell nanoparticles using the aqueous extract and dried powder of Anacardium occidentale leaf. The effects of quantity of extract/powder, temperature and pH on the formation of nanoparticles are studied. The nanoparticles are characterized using UV-vis and FTIR spectroscopies, XRD, HRTEM and SAED analyses. XRD studies show that the particles are crystalline in the cubic phase. The formation of Au core-Ag shell nanoparticles is evidenced by the dark core and light shell images in TEM and is supported by the appearance of two SPR bands in the UV-vis spectrum. FTIR spectra of the leaf powder before and after the bioreduction of nanoparticles are used to identify possible functional groups responsible for the reduction and capping of nanoparticles. Water soluble biomolecules like polyols and proteins are expected to bring about the bio-reduction.     

Sol-Gel as Reaction Matrix for Bacterial Enzymatic Activity

Sol-gel process is a rapid growing field in material chemistry. The sol-gel matrices (SGM) are basically porous wet-gels or xerogels obtained by the hydrolysis and condensation-polymerisation of metal and semimetal alkoxides, mainly SiO₂ materials. The current study presents the uses of sol-gel glass matrix (SGM) that allow direct entrapment of biomolecules within and at surface, which can be utilized by microorganisms. This glass type is solid, transparent, porous and can be modulated to form a hydrophobic or hydrophilic surface. In view of all these beneficial characteristics of SGM, preliminary data is presented on biofilm formed on thin films of SGM doped with a fluorochrome (fluorescein diacetate). The esterase/lipase activity of E. coli CN ₁₃ and K. oxytoca spp. biofilm grown on top of SGM thin film, doped with fluorescein diacetate, was detected at the level of a single cell by epifluorescence microscopy. In view of these preliminary results, sol-gel glass has a considerable potential as a variable matrix for single bacteria and biofilm investigation.     

GldA overexpressing-engineered E. coli as superior electrocatalyst for microbial fuel cells

Faster electron transfer between bacteria and electrodes in microbial fuel cells can significantly improve the power density of MFCs for practical applications. A recombinant Escherichia coli (E. coli) strain overexpressing glycerol dehydrogenase (GldA) was engineered as the MFC biocatalyst instead of the natural bacteria. Efficient mediators were produced in the fuel cell with this engineered E. coli resulting in lower polarization and much higher power density than with natural E. coli and E. coli with electro-evolved mediators. For the first time, we demonstrate that engineering E. coli by introduction of appropriate oxidoreductase via gene manipulation can greatly improve the rate of electron transfer. This work provides an efficient and economic approach to biologically engineering bacteria for improving MFC performance.     

Interactions and stability of silver nanoparticles in the aqueous phase: Influence of natural organic matter (NOM) and ionic strength

The rapid development of nanotechnology and the related production and application of nanosized materials such as engineered nanoparticles (ENP) inevitably lead to the emission of these products into environmental systems. So far, little is known about the occurrence and the behaviour of ENP in environmental aquatic systems. In this contribution, the influence of natural organic matter (NOM) and ionic strength on the stability and the interactions of silver nanoparticles (n-Ag) in aqueous suspensions was investigated using UV–vis spectroscopy and asymmetrical flow field-flow fractionation (AF⁴) coupled with UV–vis detection and mass spectrometry (ICP-MS). n-Ag particles were synthesized by chemical reduction of AgNO₃ with NaBH₄ in the liquid phase at different NOM concentrations. It could be observed that the destabilization effect of increasing ionic strength on n-Ag suspensions was significantly decreased in the presence of NOM, leading to a more stable n-Ag particle suspension. The results indicate that this behaviour is due to the adsorption of NOM molecules onto the surface of n-Ag particles (“coating”) and the resulting steric stabilization of the particle suspension. The application of AF⁴ coupled with highly sensitive detectors turned out to be a powerful method to follow the aggregation of n-Ag particle suspensions at different physical–chemical conditions and to get meaningful information on their chemical composition and particle size distributions. The method described will also open the door to obtain reliable data on the occurrence and the behaviour of other ENP in environmental aquatic systems.     

Directional persistence and the optimality of run-and-tumble chemotaxis

E. coli does chemotaxis by performing a biased random walk composed of alternating periods of swimming (runs) and reorientations (tumbles). Tumbles are typically modelled as complete directional randomisations but it is known that in wild type E. coli, successive run directions are actually weakly correlated, with a mean directional difference of 63°. We recently presented a model of the evolution of chemotactic swimming strategies in bacteria which is able to quantitatively reproduce the emergence of this correlation. The agreement between model and experiments suggests that directional persistence may serve some function, a hypothesis supported by the results of an earlier model. Here we investigate the effect of persistence on chemotactic efficiency, using a spatial Monte Carlo model of bacterial swimming in a gradient, combined with simulations of natural selection based on chemotactic efficiency. A direct search of the parameter space reveals two attractant gradient regimes, (a) a low-gradient regime, in which efficiency is unaffected by directional persistence and (b) a high-gradient regime, in which persistence can improve chemotactic efficiency. The value of the persistence parameter that maximises this effect corresponds very closely with the value observed experimentally. This result is matched by independent simulations of the evolution of directional memory in a population of model bacteria, which also predict the emergence of persistence in high-gradient conditions. The relationship between optimality and persistence in different environments may reflect a universal property of random-walk foraging algorithms, which must strike a compromise between two competing aims: exploration and exploitation. We also present a new graphical way to generally illustrate the evolution of a particular trait in a population, in terms of variations in an evolvable parameter.     

Revisiting the relationship between compositional sequence complexity and periodicity

BACKGROUND: Given a big sequence fragment or a set of functionally related sequences we consider two problems of a sequence analysis associated with the given sequence(s). The first problem is to measure sequence complexity (repetitiveness, compactness) to estimate how informative the set as a whole is. Usually an obtained measure should be compared with an appropriate random background calculated using permutation of the given sequences. We propose a novel and effective approach for background information measurement instead of the usual sequence reshuffling. The second problem is to detect a periodic bias to determine if it is one of the set features. Sequence periodicity, when sometimes one has in mind hidden periodicity, is a very basic genomic property. The sequence period of 3, which is considered to characterize coding sequences, and period 10-11, which may be due to the alternation of hydrophobic and hydrophilic amino acids, DNA curvature, and bendability were discovered and described. Searching for periodical biases brought significant results in the study of sequence-dependent nucleosome positioning: nucleosomal sites carry hidden period of about 10.4 bases. RESULTS: Calculated differences between genomic sequences and background showed high biological relevancy of the method that we proposed in this study. Our algorithm was applied to a few natural and artificial datasets. We constructed a simple "periodic" dataset by replacement of every tenth dinucleotide in each sequence of a trial set by the same dinucleotide "CC". We showed that the method reveals the introduced periodicity and that this periodical pattern carries higher information than in uninterrupted subsequences. An application of the method to the nucleosomal dataset revealed a weak pseudo-periodicity of 10.4 nucleotides confirming previous knowledge. An application of the method to Escherichia coli datasets revealed the well-known periodicity of 3bp as a genic attribute, a secondary genic period slightly larger than 11bp, and an intergenic period a bit smaller than 11bp. CONCLUSIONS: We reported a novel compositional complexity-based method for sequence analysis. We found that the difference between the sequence complexity of a natural sequence and of background is especially high for a set consisting exclusively of coding sequences. Hidden periodicities were found with no need of any preliminary assumptions regarding a composition of periodic elements. We illustrated the power of the method by studying the sets with known weak periodic properties: a nucleosomal database and sets of different regions of E. coli. We showed that the method conveniently indicated all kinds of periodicity and related features in these sets of DNA sequences.     

手性化合物组合修饰剂对CdSe纳米晶荧光性能的影响及标记大肠杆菌的研究

以手性化合物L-青霉胺、D-青霉胺、L-半胱氨酸为单一修饰剂或组合成双修饰剂,合成不同修饰剂修饰的CdSe纳米晶。对最佳合成条件如配料比,反应pH值,回流温度,回流时间进行了优化,对CdSe纳米晶发光强度及稳定性进行了系统研究。结果发现双修饰剂修饰的纳米晶比单修饰剂修饰的纳米晶荧光强度高,稳定性好;双修饰剂中第二修饰剂的空间位阻小的修饰效果好;不同手性修饰剂之间能以稳定方式结合的修饰效果好。研究了CdSe纳米晶对生物大分子的识别,仅发现核酸对CdSe纳米晶有明显的作用,用CdSe纳米晶作为荧光探针对大肠杆菌进行标记。     

Effect of pH on thermal- and chemical-induced denaturation of GFP

Green fluorescent protein (GFP) is an unusually stable autofluorescent protein that is increasingly being exploited for many applications. In this report, we have used fluorescence spectroscopy to study the effect of pH on the denaturation of GFP with sodium dodecyl sulfate (SDS), urea, and heat. Surprisingly, SDS (up to 0.5%) did not have any significant effect on the fluorescence of GFP at pH 7.5 or 8.5 buffers; however, at pH 6.5, the protein lost all fluorescence within 1 min of incubation. Similarly, incubation of GFP with 8 M urea at 50°C resulted in time dependent denaturation of GFP, but only in pH 6.5 buffer. At higher pH values (pH 7.5 and pH 8.5), the GFP was quite stable in 8 M urea at 50°C, showing only a slight decrease in fluorescence. Heat denaturation of GFP was found to be pH dependent as well, with the denaturation being fastest at pH 6.5 as compared to pH 7.5 or pH 8.5. Like the denaturation studies, renaturation of heat-denatured GFP was most efficient at pH 8.5, followed by pH 7.5, and then pH 6.5. These results suggests that GFP undergoes a structural/stability shift between pH 6.5 and pH 7.5, with the GFP structure at pH 6.5 being very sensitive to denaturation by SDS, urea, and heat.