Extracellular biosynthesis,characterization, optimization of silver nanoparticles (AgNPs) using Bacillus mojavensis BTCB15 and its antimicrobial activity against multidrug resistant pathogens

Abstract

Biosynthesis of metal nanoparticles is an area of interest among researchers because of its eco-friendly approach. Current study focuses at biosynthesis of silver nanoparticles (AgNPs) and optimization of physico-chemical conditions to obtain mono-dispersed and stable AgNPs having antimicrobial activity. Initially Bacillus mojavensis BTCB15 produced silver nanoparticles (AgNPs) of 105?nm. Silver nanoparticles (AgNPs) were characterized by particle size analyzer, UV-Vis Spectroscopy, Fourier transforms infrared spectroscopy (FTIR), Atomic force microscopy (AFM), and X-ray diffraction (XRD). Whereas, under optimal conditions of temperature 55?°C, pH 8, addition of surfactant Tween 20, and metal ion K₂SO₄, about 104% size reduction was achieved with average size of 2.3nm. Molecular characterization revealed 98% sequence homology with Bacillus mojavensis. AgNPs exhibited antibacterial activity at concentrations ranging from 0.5 to 2.5?µg/µl against Escherichia coli BTCB03, Klebsiella pneumonia BTCB04, Acinetobacter sp. BTCB05, and Pseudomonas aeruginosa BTCB01 but none against Staphylococcus aureus BTCB02. Highest antibacterial activity was observed at 0.27?µg/µl and lowest at 0.05?µg/µl of AgNPs indicated by zone of inhibition. Conclusively, under optimum conditions, Bacillus mojavensis BTCB15 was able to produce AgNPs of 2.3?nm size and had antibacterial activity against multi drug resistant pathogens.     

Polyphenolic extracts from pomegranate and watermelon wastes as substrate to fabricate sustainable silver nanoparticles with larvicidal effect against Spodoptera littoralis

The agricultural wastes adversely affect the environment; however, they are rich in polyphenols; therefore, this study aimed to employ polyphenol-enriched waste extracts for silver nanoparticles synthesis, and study the larvicidal activity of silver nanoparticles fabricated by pomegranate and watermelon peels extracts (PPAgNPs and WPAgNPs) against all larval instars of Spodoptera littoralis. The polyphenol profile of pomegranate and watermelon peel extracts (PP and WP) and silver nanoparticles was detected by HPLC. The antioxidant activity was estimated by DPPH, and FARP assays and the antimicrobial activity was evaluated by disc assay. The Larvicidal activity of AgNPs against Egyptian leaf worm was performed by dipping technique. The obtained AgNPs were spherical with size ranged 15–85 nm and capped with proteins and polyphenols. The phenolic compounds in silver nanoparticles increased about extracts; therefore, they have the best performance in antioxidant/reducing activity, and inhibit the growth of tested bacteria and yeast. The PPAgNPs were the most effective against the first instar larvae instar (LC₅₀ = 68.32 µg/ml), followed by pomegranate extract with (LC₅₀ = 2852 µg/ml). The results indicated that obvious increase in polyphenols content in silver nanoparticles enhance their larvicidal effect and increasing mortality of 1^st larval of S. littoralis Egyptian leafworms causing additive effect and synergism. We recommend recycling phenolic enriched agricultural wastes in producing green silver nanoprticles to control cotton leafworm that causes economic loses to crops.     

Comparative study of antifungal activity of two preparations of green silver nanoparticles from Portulaca oleracea extract

The green silver nanoparticles (green AgNPs) exhibit an exceptional antimicrobial property against different microbes, including bacteria and fungi. The current study aimed to compare the antifungal activities of both the crude aqueous extract of Portulaca oleracea or different preparations of green AgNPs biosynthesized by mixing that aqueous extract with silver nitrate (AgNO₃). Two preparations of the green AgNPs were synthesized either by mixing the aqueous extract of P. oleracea with silver nitrate (AgNO₃) (normal AgNPs) or either irradiation of the AgNPs, previously prepared, under ⁶⁰Co γ-ray using chitosan (gamma-irradiated AgNPs). Characterization of different AgNPs were tested by Zeta potential analyzer, Ultraviolet (UV) Visible Spectroscopy, and Fourier-Transform Infrared (FTIR) spectrometry. Three different plant pathogenic fungi were tested, Curvularia spicifera, Macrophomina phaseolina, and Bipolaris sp. The antifungal activities were evaluated by Transmission Electron Microscope (TEM) for either the crude aqueous extract of P. oleracea at three doses (25%, 50%, and 100%) or the newly biosynthesized AgNPs, normal or gamma-irradiated. With a few exceptions, the comparative analysis revealed that the irradiated green AgNPs at all three concentrations showed a relatively stronger antifungal effect than the normal AgNPs against all the three selected fungal strains. UV–visible spectroscopy of both preparations showed surface plasmon resonance at 421 nm. TEM results showed that both AgNPs were aggregated and characterized by a unique spherical shape, however, the gamma-irradiated AgNPs were smaller than the non-irradiated AgNPs (0.007–0.026 µM vs. 0.009–0.086 µM). TEM photographs of the fungal strains treated with the two AgNPs preparations showed flaccid structures, condensed hyphae, and shrunken surface compared with control cells. The data suggested that the biosynthesized P. oleracea AgNPs have antifungal properties against C. spicifera, M. phaseolina, and Bipolaris sp. These AgNPs may be considered a fungicide to protect different plants against phytopathogenic fungi.     

Green synthesis of silver nanoparticles using Citrus limon peels and evaluation of their antibacterial and cytotoxic properties

The present work aimed to synthesis silver nanoparticles (AgNPs) using biological waste products Citrus limon peels, its characterization, antimicrobial activities and the cytotoxic effect of the synthesized green AgNPs. Characterization of the prepared AgNPs showed the formation of spherical, and few agglomerated AgNPs forms as measured by UV–visible spectrophotometer. The average size of the prepared AgNPs was 59.74 nm as measured by DLS technique. The spectrum of the synthesized AgNPs was observed at 3 KeV using the EDX. On the other hand, FTIR analysis of the green synthesized AgNPs showed the presence of alcohols, phenolics, mono-substituted alkynes, aliphatic primary amines, sodium salt, amino acid, or SiOH alcohol groups. The antimicrobial studies of the formed AgNPs showed positive activity against most of the studied human pathogenic bacteria with varying degrees. Finally, the evaluation of the cytotoxic effect of the green synthesized AgNPs were done using two types of cell lines, human breast cancer cell line (MCF-7) and human colon carcinoma cell line (HCT-116). The results revealed the concentration has a direct correlation with cell viability. The 50% inhibitory concentration (IC₅₀) of MCF-7 cell line was in of 23.5 ± 0.97 µL/100 µL, whereas the HCT-116 cell line was in 37.48 ± 5.93 µL/100 µL.     

Antioxidant,antibacterial and cytotoxic potential of silver nanoparticles synthesized using terpenes rich extract of Lantana camara L. leaves

Several attempts have been made for green synthesis of silver nanoparticles (AgNPs) using different plant extracts. Present study revealed that, antioxidant, antibacterial and cytotoxic AgNPs were synthesized using terpenes-rich extract (TRE) of environmentally notorious Lantana camara L. leaves. AgNPs were characterized by advanced techniques like UV–Visible and Infra red spectroscopy; XRD, SEM techniques as terpenes coated sphere shaped NPs with average diameter 425 nm. Further, on evaluation, AgNPs were found to exhibit dose – dependent antioxidant potential, good to moderate antibacterial activity against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa; and toxicity on Brine shrimp (A. salinanauplii) with LD₅₀ value 514.50 µg/ml.     

Novel marine Nocardiopsis dassonvillei-DS013 mediated silver nanoparticles characterization and its bactericidal potential against clinical isolates

The sediment marine samples were obtained from several places along the coastline of the Tuticorin shoreline, Tamil Nadu, India were separated for the presence of bioactive compound producing actinobacteria. The actinobacterial strain was subjected to 16Sr RNA sequence cluster analysis and identified as Nocardiopsis dassonvillei- DS013 NCBI accession number: KM098151. Bacterial mediated synthesis of nanoparticles gaining research attention owing its wide applications in nonmedical biotechnology. In the current study, a single step eco-friendly silver nanoparticles (AgNPs) were synthesized from novel actinobacteria Nocardiopsis dassonvillei- DS013 has been attempted. The actinobacterial mediated silver nanoparticles were characterized by TEM, UV–Visible, XRD, FT-IR spectroscopy. The initial detection of AgNPs was identified using UV–Vis spectrum and confirmed by the appearance of absorbance peak at 408 nm. A Fourier transform infrared spectroscopy (FT-IR) result reveals the presence of protein component in the culture supernatant may act as protecting agents. The XRD pattern indicated that the typical peaks reveal the presence of nanoparticles. The TEM morphology confirms the formation of circular and non uniform distributions of AgNPs with the size range from 30 to 80 nm. The antibacterial activity of both isolated actinobacterial (IA) and silver nanoparticles mediated actinobacterial (SNA) of Nocardiopsis dassonvillei- DS013 were done by well diffusion method against selected clinical isolates of bacteria, namely Escherichia coli, Enterococcus sp., Pseudomonas sp., Klebsiella sp., Proteus sp., Shigella sp., Bacillus subtilis, and Streptococcus sp. When compared to isolated actinobacteria, the SNA shows the better antibacterial activity against clinical isolates.     

Antifungal Susceptibility Profile of Candida Albicans Isolated from Vulvovaginal Candidiasis in Xinjiang Province of China

We investigated the antifungal susceptibility profiles of 207 independent Candida albicans strains isolated from patients with vulvovaginal candidiasis (VVC) in Xinjiang Province of China. Using CLSI M27-A3 and M27-S4 guidelines, anidulafungin and micafungin were the most active drugs against C. albicans showing an MIC₅₀/MIC₉₀ corresponding to 0.016/0.0313 µg/mL, followed by caspofungin (0.25/0.25 µg/mL), posaconazole (0.125/0.5 µg/mL), ravuconazole (0.063/1 µg/mL), itraconazole (0.125/1 µg/mL), amphotericine B (0.5/1 µg/mL), isavuconazole (0.063/2 µg/mL), 5-flucytosine (1/2 µg/mL), voriconazole (0.125/4 µg/mL), and fluconazole (0.5/4 µg/mL). 96.1% (199)–100.0% (207) isolates were sensitive to the three echinocandins tested, amphotericine B and 5-flucytosine. The in vitro activity of triazoles against all isolates tested was variable; itraconazole and voriconazole had reduced the activity to almost half of the isolates (55.1% (114) and 51.2% (106) susceptible, respectively). Fluconazole was active against 76.3% (158) isolates tested. The new triazoles ravuconazole, isavuconazole and posaconazole showed good in vitro potency against 89.9% (186)–95.2% (197) of isolates with the geometric mean MIC (µg/mL) of 0.10, 0.12 and 0.14 µg/mL, respectively. In conclusion, our study indicates that for effective management of systemic candidiasis in Xinjiang Province of China, it is important to determine the susceptibility profiles of isolated C. albicans from patients with VVC.

     

Green biosynthesis of silver nanoparticles from Annona squamosa leaf extract and its in vitro cytotoxic effect on MCF-7 cells

The biological method for the synthesis of silver nanoparticles (AgNPs) using Annona squamosa leaf extract and its cytotoxicity against MCF-7 cells are reported. The synthesized AgNPs using A. squamosa leaf extract was determined by UV–visible spectroscopy and it was further characterized by FT-IR, X-ray diffraction (XRD), Transmission electron microscopy (TEM), Zeta potential and energy dispersive spectrometric (EDS) analysis. The UV–visible spectrum showed an absorption peak at 444 nm which reflects surface plasmon resonance (SPR) of AgNPs. TEM photography showed biosynthesized AgNPs were predominantly spherical in shape with an average size ranging from 20 to 100 nm. The Zeta potential value of ?37 mV revealed the stability of biosynthesized AgNPs. Furthermore, the green synthesized AgNPs exhibited a dose-dependent cytotoxicity against human breast cancer cell (MCF-7) and normal breast epithelial cells (HBL-100) and the inhibitory concentration (IC₅₀) were found to be 50 μg/mL, 30 μg/mL, and 80 μg/mL, 60 μg/ml for AgNPs against MCF-7 and normal HBL-100 cells at 24 h and 48 h incubation respectively. An induction of apoptosis was evidenced by (AO/EtBr) and DAPI staining. Application of such eco-friendly nanoparticles makes this method potentially exciting for the large scale synthesis of nanoparticles.     

Phytosynthesis of silver nanoparticles from Jatropha integerrima Jacq. flower extract and their possible applications as antibacterial and antioxidant agent

Jatropha integerrima Jacq. flower extract was used for the synthesis of silver nanoparticles in the current study. Various spectroscopic analyses were used to characterize the synthesized nanoparticles (JIF-AgNPs). The antibacterial efficacy of JIF-AgNPs was studied by well diffusion and microdilution techniques. In addition, the impact of JIF-AgNPs on free radicals was evaluated. On the ultraviolet–visible spectrum, the nanoparticles exhibit the highest absorbance at 422 nm. Based on the Fourier transform infrared spectrum, phenols and amino acids were involved in capping the JIF-AgNPs. Crystalline sphere-shaped nanoparticles with an average size of 50.07 nm and zeta potential of ?19.0 mV were confirmed by X-ray diffraction, transmission electron microscopy, and dynamic light scattering analysis respectively. The JIF-AgNPs exhibit the highest and lowest growth inhibitory activity towards E. coli and B. subtilis. The minimal inhibitory concentration of JIF-AgNPs against E. coli, K. pneumoniae, S. aureus, and B. subtilis were 2.5, 5.0, 5.0, and 7.5 μg/mL, respectively. The JIF-AgNPs exhibited significant radical scavenging activities against DPPH (IC₅₀-32.5 ± 0.06 µg/mL), hydroxyl (IC₅₀-25 ± 0.09 µg/mL), Superoxide (IC₅₀-42.5 ± 0.13 µg/mL), and ABTs (IC₅₀-33.5 ± 0.15 µg/mL). Thus, synthesized nanoparticles were a good alternative to develop an antibacterial and antioxidant agent.     

Fluorimetric determination of proteins using 4‐chloro‐(2′‐hydroxylophenylazo)rhodanine–Ti(IV) complex as a spectral probe

A novel method for the determination of proteins was developed, based on the enhancement of fluorescence with 4‐chloro‐(2′‐hydroxylophenylazo)rhodanine–Ti(IV) [ClHARP–Ti(IV)] complex as a fluorescence probe. The excitation and emission wavelengths of the system were 335 nm and 376 nm, respectively. The presence of bis(2‐ethylhexyl)sulphosuccinate sodium salt (AOT) microemulsion greatly increased the sensitivity of the system. Under optimal conditions, four kinds of proteins, including bovine serum albumin (BSA), human serum albumin (HSA), egg albumin (Ova), and γ‐globin (γ‐G) were studied. The detection limits were 0.182 µg/mL for BSA, 0.0788 µg/mL for HSA, 0.216 µg/mL for Ova and 0.484 µg/mL for γ‐G. The linear ranges of the calibration were 0–12.0, 0–10.0, 0–18.0 and 0–18.0 µg/mL, respectively. The method possessed high sensitivity, good selectivity and was applied to the analysis of protein in milk powder and cornmeal with satisfactory results. Copyright © 2008 John Wiley & Sons, Ltd.     

Pharmacological properties of biogenically synthesized silver nanoparticles using endophyte Bacillus cereus extract of Berberis lyceum against oxidative stress and pathogenic multidrug-resistant bacteria

The emergence of multidrug resistance in pathogenic bacteria limits the utilization of available antibiotics. The development of alternate options to treat infectious diseases is the need of the day.The present study was aimed to synthesize, characterize and evaluate the bioactive properties of silver nanoparticles. Endophytic bacterium Bacillus cereus (MT193718) isolated from Berberis lycium was used to synthesize biocompatible silver nanoparticles. Antibacterial properties of AgNPs were evaluated against clinically isolated multidrug-resistant strains of Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae. AgNPs indicated significant antibacterial activity against S. aureus and K. pneumoniae fwith a zone of inhibition of 17 and 18 mm at a concentration of 1000 µg/ mL with minimum inhibitory concentration of 15.6 and 62.5 µg/mL respectively. Significant antioxidant activity with an IC50 value of 9.5 µg/mL was recorded. Biosynthesized AgNPs were found compatible with red blood cells at a concentration of 31.5 µg/ml with no clumping of erythrocytes. The study suggested that AgNPs synthesized by the endophytic bacterium Bacillus cereus are biologically active and can be used as antioxidant and antibacterial agents against drug-resistant bacteria.     

Biosynthesis of silver nanoparticles by a new strain of <Emphasis Type="Italic">Streptomyces</Emphasis> sp. compared with <Emphasis Type="Italic">Aspergillus</Emphasis><Emphasis Type="Italic">fumigatus</Emphasis>

Locally isolated strains of a thermoalkalotolerant Streptomyces sp. and Aspergillus fumigatus were used for the in vitro biosynthesis of silver nanoparticles from AgNO₃ solutions. An autolysed cell-free culture filtrate from each strain was used, indicating that the formation mechanism depends on intra-cellular components for both organisms, since culture broths had no significant nanoparticle formation potential. Nanoparticle formation was indicated by a change of the solution from colourless or light brown to dark brown after 24 h or more, and UV–visible spectroscopy and x-ray diffraction analysis confirmed the formation by both organisms. The initial formation kinetics were faster with Aspergillus, but formation continued for a longer period with Streptomyces, resulting in higher concentrations after 48 h. Transmission electron microscope images revealed well dispersed nanoparticles with diameters ranging from 15 to 45 nm from A. fumigatus, while those from Streptomyces sp. had a narrower size distribution of 15–25 nm. The higher productivity and preferred narrower size distribution of Streptomyces, together with its well established industrial use, may make it the preferred choice for further optimization studies.     

Simultaneous bioremediation of Cr(VI) and lindane in soil by actinobacteria

Environments co-contaminated with metals and organic compounds are difficult to remediate. Actinobacteria is an important group of microorganisms found in soils, with high metabolic versatility and potential for bioremediation. In this paper, actinobacteria were used to remediate soil co-contaminated with Cr(VI) and lindane. Five actinobacteria, tolerant to Cr(VI) and lindane mixture were selected: Streptomyces spp. A5, A11, M7, and MC1, and Amycolatopsis tucumanensis DSM 45259. Sterilized soil samples were inoculated with actinobacteria strains, either individually or as a consortium, and contaminated with Cr(VI) and lindane, either immediately or after 7 days of growth, and incubated at 30 °C during 14 days. All actinobacteria were able to grow and remove both contaminants, the consortium formed by Streptomyces spp. A5, M7, MC1, and A. tucumanensis showed the highest Cr(VI) removal, while Streptomyces sp. M7 produced the maximum lindane removal. In non-sterile soil samples, Streptomyces sp. M7 and the consortium removed more than 40% of the lindane, while Streptomyces sp. M7 demonstrated the greatest Cr(VI) removal. The most appropriate strategy for bioremediation of Cr(VI) and lindane co-contaminated soils would be the inoculation with Streptomyces sp. M7.     

Prospective of biosynthesized L.satiVum oil/PEG/Ag-MgO bionanocomposite film for its antibacterial and anticancer potential

A substantial interest has been manifested in utilizing oil/metal oxide hybrid bionanocomposite, especially organic/ inorganic to design different biomedical applications. The present study reports the synthesis, characterization, antibacterial and anticancer properties of biogenic silver nanoparticles (AgNPs) and L.satiVum oil/PEG/Ag-MgO bionanocomposite. The fabricated AgNPs and L.sativum oil/PEG/Ag-MgO bionanocomposite were characterized by employing different spectroscopic (UV, FTIR, XRD) and microscopic (TEM, SEM) techniques. The particle size analysis showed that the mean size of 16.32 nm for AgNPS and 13.45 nm L.satiVum oil/PEG/Ag-MgO, indicating the excellent dispersion of Ag-MgO nanoparticles in the PEG– L.satiVum oil matrix. The antimicrobial activity of AgNPs and polymeric bionanocomposite was investigated against two pathogenic bacteria. The highest antibacterial effect was observed for bionanocomposite towards Gram-positive Staphylococcus aureus (27 mm) and Gram-negative Escherichia coli (25 mm) at 40 µg/well. The bionanocomposite completely vanished the bacterial growth (100%) at 80 µgmL⁻¹ concentrations. Moreover, the AgNPs and polymeric bionanocomposite was evaluated for anticancer activity against human cervical cancer cells (HeLa cells) at different doses (50, 250, 500, and 1000 µgmL⁻¹). The results showed polymeric bionanocomposite was stronger in inducing the HeLa cancer cell death than AgNPs. Overall, the fabricated L.satiVum oil/PEG/Ag-MgO bionanocomposite serve as a potential antimicrobial and anticancer agent and could be used in the development of novel drugs and health care products in near future.     

Bioprospecting of the novel isolate Microbacterium proteolyticum LA2(R) from the rhizosphere of Rauwolfia serpentina

The study aimed to assess the proficiency of secondary metabolites (SMs) synthesized by actinobacteria isolated from the rhizospheric soil of Rauwolfia serpentina for its antimicrobial and anti-biofilm activity. After morphological and biochemical identification of actinobacteria, primary and secondary screening was done for specific metabolite production. The secondary metabolites were then tested for their antioxidant, antibacterial, and antibiofilm potential. Out of 29 bacterial colonies isolated, only one emerged as a novel isolate, Microbacterium LA2(R). Partial 16S rRNA gene sequence of the isolate LA2(R) was deposited in NCBI GenBank with accession number MN560041. The highest antioxidant capacity of the methanolic extract the novel isolate was found to be 474.183 µL AAE/mL and 319.037 µL AAE/mL by DPPH assay and ABTS assay respectively; three folds higher than the control. These results were further supported by the high total phenolic (194.95 gallic acid equivalents/mL) and flavonoid contents (332.79 µL quercetin equivalents/mL) of the methanolic extract. GC–MS analysis revealed the abundance of antibacterial compounds; where, n-Hexadecanoic acid was found to be the major compound present with a peak of 14 min retention time (RT) and 95% similarity index. MIC value of the metabolite was noted to be around 132.28 ± 84.48 μg/mL. The IC₅₀ value was found to be 74.37, 71.33, 66.28 and 84.48 μg/mL against Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia, and Salmonella abony, respectively. Treatment with IC₅₀ of the extract decreased the biofilm formation up to 70%–80% against pathogenic strains viz. Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae and Salmonella abony. These significant activities of Microbacterium sp. LA2(R) suggests that it could be utilized for antibiotic production for human welfare and in various important industrial applications.     

Phytochemical analysis and fabrication of silver nanoparticles using Acacia catechu: An efficacious and ecofriendly control tool against selected polyphagous insect pests

Globally, the farmers are struggling with polyphagous insect pest, and it is the number one enemy of agri-products, which made plenty of economic deterioration. Spodoptera litura and Helicoverpa armigera are the agronomically important polyphagous pests. Most of the farmers are predominately dependent on synthetic chemical insecticides (SCIs) for battle against polyphagous pets. As a result, the broad spectrum usage of SCIs led a lot of detrimental outcomes only inconsequently the researchers search the former-friendly phyto-pesticidal approach. In the present investigation, leaf ethanol extract (LEE) and silver nanoparticles (AgNPs) of A. catechu (Ac) were subjected to various spectral (TLC, CC, UV, FTIR, XRD and SEM) analyses. Larval and pupal toxicity of A. catechu Ac-LEE and Ac-AgNPs were tested against selected polyphagous insect pests. The significant larval and pupal toxicity were experimentally proven, and the highest toxicity noticed in AgNPs than Ac-LEE. The larval and pupal toxicity of Ac-AgNPs tested against S. litura and H. armigera LC₅₀/LC₉₀ values were 71.04/ 74.78, 85.33/ 88.91 µg/mL and 92.57/ 96.21 and 124.43/ 129.95 µg/mL respectively. Ac-AgNPs could be potential phyto-pesticidal effectiveness against selected polyphagous insect pests. In globally, it is significantly sufficient ratification giving towards the prevention of many unauthorized SCPs.     

Characterization of a thermostable and solvent-tolerant laccase produced by Streptomyces sp. LAO

Objectives

Decaying wood samples were collected, and actinomycetes were isolated and screened for laccase production. The identity of the efficient laccase-producing isolate was confirmed by using a molecular approach. Fermentation conditions for laccase production were optimized, and laccase biochemical properties were studied.

Results

Based on the 16S rRNA gene sequencing and phylogenetic analysis, the isolate coded as HWP₃ was identified as Streptomyces sp. LAO. The time-course study showed that the isolate optimally produced laccase at 84 h with 40.58?±?2.35 U/mL activity. The optimized physicochemical conditions consisted of pH 5.0, ferulic acid (0.04%; v/v), pine back (0.2 g/L), urea (1.0 g/L), and lactose (1 g/L). Streptomyces sp. LAO laccase was optimally active at pH and temperature of 8.0 and 90 °C, respectively, with remarkable pH and thermal stability. Furthermore, the enzyme had a sufficient tolerance for organic solvents after 16 h of preincubation, with laccase activity?>?70%. Additionally, the laccase maintained considerable residual activity after pretreatment with 100 mM of chemical agents, including sodium dodecyl sulphate (69.93?±?0.89%), ethylenediaminetetraacetic acid (93.1?±?7.85%), NaN₃ (96.28?±?3.34%) and urea (106.03?±?10.72%).

Conclusion

The laccase's pH and thermal stability; and robust catalytic efficiency in the presence of organic solvents suggest its industrial and biotechnological application potentials for the sustainable development of green chemistry.

     

Anticancer,antimicrobial, antioxidant,and catalytic activities of green-synthesized silver and gold nanoparticles using Bauhinia purpurea leaf extract

The synthesis of metal nanoparticles by green methods attained enormous attention in recent years due to its easiness, non-toxicity, and eco-friendly nature. In the present study, noble metal nanoparticles such as silver and gold were prepared using an aqueous leaf extract of a medicinal plant, Bauhinia purpurea. The leaf extract performed as both reducing and stabilizing agents for the development of nanoparticles. The formations of silver and gold nanoparticles were confirmed by observing the surface plasmon resonance peaks at 430 nm and 560 nm, respectively, in UV–Vis absorption spectrum. Various properties of nanoparticles were demonstrated using the characterization techniques such as FTIR, XRD, TEM, and EDX. The synthesized silver and gold nanoparticles had a momentous anticancer effect against lung carcinoma cell line A549 in a dose-dependent manner with IC₅₀ values of 27.97 µg/mL and 36.39 µg/mL, respectively. The antimicrobial studies of synthesized nanoparticles were carried out by agar well diffusion method against six microbial strains. Silver and gold nanoparticles were also showed high antioxidant potentials with IC₅₀ values of 42.37 µg/mL and 27.21 µg/mL, respectively; it was measured using DPPH assay. Additionally, the nanoparticles were observed to be good catalysts for the reduction of organic dyes.

     

Trace analysis of N‐acetyl‐L‐cysteine using luminol–H2O2 chemiluminescence system catalyzed by silver nanoparticles

N‐Acetyl‐L‐cysteine (NAC) can inhibit the luminol–H₂O₂, reaction, which is catalyzed by silver nanoparticles. Based on this phenomenon a new method was developed for NAC determination. Under optimum conditions, a linear relationship between chemiluminescence intensity and NAC concentration was found in the range 0.034–0.98 µg/mL. The detection limit was 0.010 µg/mL (S/N =3), and the relative standard deviation (RSD) was <5% for 0.480 µg/mL NAC (n =5). This simple, sensitive and inexpensive method has been applied to measure the concentration of NAC in pharmaceutical tablets. Copyright © 2014 John Wiley & Sons, Ltd.     

Novel biosynthesis,characterization and bio-catalytic potential of green algae (Spirogyra hyalina) mediated silver nanomaterials

In recent years green nanotechnology gained significant importance to synthesize nanoparticles due to their cost effectiveness and biosafety. In the current study, silver nanoparticles were synthesized by using extract of Spirogyra hyalina as a capping and reducing agent. The synthesized nanoparticles were characterized by UV–Visible spectroscopy, Fourier transform infrared spectroscopy, Scanning electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffractive analysis. Silver nanoparticles give a characteristic Surface Plasmon Resonance peak of 451 nm at 2.21 a.u (arbitrary unit). SEM micrograph revealed the spherical morphology and average grain size of 52.7 nm. Furthermore, antibacterial, antifungal, insecticidal, antioxidant and membrane damage activities were determined. The maximum antibacterial and antifungal activity was observed for Pseudomonas aeruginosa (18 ± 1.2 mm) and Fusarium solani (14.3 ± 0.6 mm), respectively. In membrane damage assay, Pseudomonas aeruginosa absorbed A₂₆₀ wavelength and gave maximum peak values of 0.286, 0.434 and 0.629 at 25, 35 and 45 µg/mL of silver nanoparticles. The membrane damage assay confirmed that nanoparticles are involved in bacterial cell membrane damage. At 500 ppm silver nanoparticles showed 30% mortality against Tribolium castaneum (a common grain pest). The silver nanoparticles also showed potent antioxidant activity and successfully scavenged the DPPH free radicals upto 53.43 ± 0.17, 43.26 ± 0.97, 31.39 ± 0.33, 24.62 ± 0.85, and 14.13 ± 0.12% at a concentration of 400, 200, 100, 50, and 25 µg/mL of nanoparticles, respectively. It is concluded that silver nanoparticles can easily be synthesized by using green algae Spirogyra hyalina as a capping and reducing agent. Silver nanoparticles showed potent biomedical activities and thus can be used for therapeutic applications invitro and invivo.