Silver and gold nanoparticles: Eco-friendly synthesis, antibiofilm, antiviral, and anticancer bioactivities.

For the first time in this study, silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) were green synthesized by the cost-effective and eco-friendly procedure using Cotton seed meal and Fodder yeast extracts. The biosynthesized NPs were characterized by UV-Vis spectroscopy, dynamic light scattering analysis (DLS), transmission electron microscopy (TEM), selected area electron diffraction (SAED), and fourier-transform infrared (FTIR) spectroscopy. Furthermore, the biosynthesized NPs were tested in vitro against biofilm formation by some pathogenic negative bacteria (Escherichia coli, Proteus mirabilis, Klebsiella sp., Salmonella sp., and Pseudomonas aeruginosa) and negative bacteria (staphylococcus aureus) as well as against human denovirus serotype 5 (HAdV-5) and anticancer activity using HepG2 hepatocarcinoma cells. UV-Vis absorption spectra of reaction mixture of AgNPs and AuNPs exhibited maximum absorbance at 440 nm and 540 nm, respectively. This finding was confirmed by DLS measurements that the highest intensity of the AgNPs and AuNPs were 84 nm and 73.9 nm, respectively. FTIR measurements identified some functional groups detected in Cotton seed meal and Fodder yeast extracts that could be responsible for reduction of silver and gold ions to metallic silver and gold. The morphologies and particle size of AgNPs and AuNPs were confirmed by the TEM and SAED pattern analysis. Biosynthesized AgNPs and AuNPs showed good inhibitory effects against biofilms produced by Escherichia coli, Proteus mirabilis, Klebsiella sp., Salmonella sp., Pseudomonas aeruginosa, and Staphylococcus aureus. In addition, they showed anticancer activities against hepatocellular carcinoma (HepG-2) and antiviral activity against human adenovirus serotype 5 infection in vitro. Finally, the results of this study is expected to be extremely helpful to nano-biotechnology, pharmaceutical, and food packing applications through developing antimicrobial and/or an anticancer drugs from ecofriendly and inexpensive nanoparticles with multi-potentiality.

Efficient malachite green biodegradation by Pseudomonas plecoglossicide MG2: process optimization, application in bioreactors, and degradation pathway.

Microbial degradation of synthetic dyes is considered a promising green dye detoxification, cost-effective and eco-friendly approach. A detailed study on the decolorization and degradation of malachite green dye (MG) using a newly isolated Pseudomonas plecoglossicide MG2 was carried out. Optimization of MG biodegradation by the tested organism was investigated by using a UV-Vis spectrophotometer and the resultant degraded products were analyzed by liquid chromatography-mass spectrometry and FTIR. Also, the cytotoxicity of MG degraded products was studied on a human normal retina cell line. The optimum conditions for the significant maximum decolorization of MG dye (90-93%) by the tested organism were pH 6-7, inoculum size 4-6%, and incubation temperature 30-35 °C, under static and aerobic conditions. The performance of Pseudomonas plecoglossicide MG2 grown culture in the bioreactors using simulated wastewater was assessed. MG degradation (99% at 100 and 150 mg MG/l at an optimal pH) and COD removal (95.95%) by using Pseudomonas plecoglossicide MG2 culture were the best in the tested culture bioreactor in comparison with that in activated sludge or tested culture-activated sludge bioreactors.The FTIR spectrum of the biodegraded MG displayed significant spectral changes, especially in the fingerprint region 1500-500 as well as disappearance of some peaks and appearance of new peaks. Twelve degradation intermediates were identified by LC-MS. They were desmalachite green, didesmalachite green, tetradesmalachite green, 4-(diphenylmethyl)aniline, malachite green carbinol, bis[4-(dimethylamino)phenyl]methanone, [4-(dimethylamino)phenyl][4-(methyl-amino)phenyl]methanone, bis[4-(methylamino)phenyl]methanone, (4-amino- phenyl)[4-(methylamino)phenyl]methanone, bis(4-amino phenyl)methanone, (4-amino phenyl)methanone, and 4-(dimathylamino)benzaldehyde. According to LC-MS and FTIR data, two pathways for MG degradation by using Pseudomonas plecoglossicide MG2 were proposed. MG showed cytotoxicity to human normal retina cell line with LC50 of 28.9 µg/ml and LC90 at 79.7 µg/ml. On the other hand, MG bio-degraded products showed no toxicity to the tested cell line. Finally, this study proved that Pseudomonas plecoglossicide MG2 could be used as an efficient, renewable, eco-friendly, sustainable and cost-effective biotechnology tool for the treatment of dye wastewater effluent.

Modeling and in-vivo evaluation of fibrinolytic enzyme produced by Bacillus subtilis Egy under solid state fermentation.

Blood clot formation increases cases of myocardial infarction (AMI) and stroke, thus urges directing much research works for treatment and prevention of the causes. One of these directions is the microbial production of fibrinolytic enzymes as thrombolytic agents. In the current work, Bacillus subtilis Egy has been used for enzyme production under solid state fermentation. Among twelve nutrient meals in addition to wheat bran as a control fodder yeast yielded the highest enzyme activity reaching 114U/g. Applying statistical model for optimization of enzyme production revealed that 3.6%, fodder yeast; 40%, moisture content; 6 days, incubation period and 2%, inoculum size were the optimum conditions for maximum fibrinolytic enzyme production (141.02 U/g) by Bacillus subtilis Egy under solid-state fermentation The model was significant and data were experimentally validated. The produced fibrinolytic enzyme was evaluated for in vitro and in vivo cytotoxicity. In-vivo examination of the enzyme resulted in no mortality during the first 24 h after treatment. After 14 days, the results revealed no significant changes detected in hematological parameters (RBCs, MCV, hemoglobin except WBCs which showed an increase for both sexes. Histopathological examination of liver and kidney of rats received oral and subcutaneous treatments showed normal architecture. The data showed the applicability of the produced enzyme for the treatment of blood clot with no significant effect on living cells or on physiological functions.

Optimization of Bacillus subtilis growth parameters for biosynthesis of silver nanoparticles by using response surface methodology.

Silver nanoparticles (AgNPs) are among the most widely biosynthesized and used nanomaterials. They have different unique properties and a wide range of applications. This study is concerned with optimization of the growth conditions of Bacillus subtilis NRC1 for the biosynthesis of AgNPs using two designs of response surface methodology (RSM) statistical analysis. The data obtained from Plackett-Burman design (PBD) followed by central composite design (CCD), showed a good agreement between the experimental and predicted values of AgNPs biosynthesis. The optimum conditions were 0.7% (w/v) casein hydrolysate, 5% dextrin (w/v), pH 7.5 and 57 × 106 CFU/ml inoculum size. The model was highly valid and could be applied with a confidence factor of 99.47%. Minimal inhibitory concentration (MIC) of these AgNPs synthesized using the extracellular filtrate after growth of Bacillus subtilis NRC1 in the optimized medium was found to be 41-43µg/ml for all tested microorganisms with exception of Pseudomonas aeruginosa where MIC was 169 µg/ml.

Optimization of Bacillus subtilis NRC1 growth conditions using response surface methodology for sustainable biosynthesis of gold nanoparticles.

Gold nanoparticles (AuNPs) have different unique properties and a wide range of applications in different fields. Thereby, there is a growing urgency for the production of AuNPs using a safe and an economic method. In this study, optimization of fermentation conditions by Bacillus subtilis NRC1 for extracellular AuNPs synthesis using response surface methodology was achieved. The data obtained from Plackett-Burman design followed by Box-Behnken design indicated the accuracy and reliability of the model and it could be used to navigate the design space with a reasonable accuracy. Numerical optimization of Bacillus subtilis NRC1 active extracellular filtrate production, showed the optimum conditions of 0.74% (w/v) casein hydrolysate, 3.99% (w/v) dextrin, 47 × 106 CFU/ml inoculum size at pH 7.76 and 25 [Formula: see text]C to give the maximum AuNPs biosynthesis. The model was highly valid and the obtained data had a confidence factor of 98.48%. Statistical optimization resulted in a 2.6-fold increase in AuNPs production compared with that of the non-optimized medium.

Potential of silver nanoparticles synthesized using low active mosquitocidal Lysinibacillus sphaericus as novel antimicrobial agents.

Silver nanoparticles (AgNPs) were synthesized using extracellular filtrates of some Lysinibacillus sphaericus (Ls) strains under simple conditions. Ls synthesized AgNPs showed the optical absorption peaks at 388-412 nm as detected by UV-visible spectrophotometer. Transmission electron micrographs of bacterial synthesized AgNPs revealed that they were polycrystalline with spherical, hexagonal, cuboidal, rod and irregular shapes. The average diameter of the tested AgNPs were ranged from 14-21 nm and they were negatively charged as detected by DLS (-18.2 to -28.9). FTIR spectra showed the presence of nitrogenous biomolecules capping the synthesized AgNPs. The filtrates of tested Ls strains showed nitrate reductase activity (1.45-2.56 µmol/ml/min). Tested AgNPs showed bactericidal activity against Gram positive and Gram negative bacteria, fungicidal activity against yeast and filamentous fungi, and virucidal activity against rotavirus. In addition, it showed synergistic antimicrobial effect to cephradine and nizoarm against all tested microorganisms. Cytotoxicity test revealed the safety of the tested nanoparticles at tested concentrations.Finally, Ls strains represent microbial sources for ecofriendly, simple and economic biosynthesis of antimicrobial AgNPs. Also, this research may contribute to the medicinal chemistry and pharmaceutical industry for the development of new products used for the public health.

Biosynthesis of silver nanoparticles using isolated Bacillus subtilis: characterization, antimicrobial activity, cytotoxicity, and their performance as antimicrobial agent for textile materials.

Silver nanoparticles (AgNPs) have unique properties and a large range of applications. Biosynthesis of stable AgNPs using the extracellular filtrate of Bacillus subtilis was proved by the characteristic surface plasmon resonance at about 420-430 nm. They were polycrystalline with spherical, hexagonal, and irregular shapes and they were negatively charged (-40 mV) with an average diameter of 20 nm. FTIR spectrum confirmed the presence of protein molecules coating AgNPs. The optimum conditions for the synthesis of tested AgNPs were 1:6 filtrate dilution, 1 mM AgNO3, pH 7, 30 °C , 48 h contact time under static and illuminating conditions. The synthesized AgNPs showed antibacterial activities against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus, and Salmonella typhi, antifungal activity against Candida albicans and antiviral activity against rotavirus. Also, they showed potent cytotoxic effects on lung and hepatic carcinoma human cell lines. Meanwhile, the acute toxicity study against mice showed no significant changes in hematological, biochemical, and histological parameters of AgNPs treated mice. They also showed mild hepatoprotective effect in thioacetamide (TAA) - induced hepatic fibrosis in rats. AgNPs treated textiles fabrics showed high antimicrobial activities against different pathogenic microorganisms as well as UV protection adequacy.

Optimization of spore laccase production by Bacillus amyloliquefaciens isolated from wastewater and its potential in green biodecolorization of synthetic textile dyes.

The spore laccase enzyme production by B. amyloliquefaciens was optimized. It was characterized and tested for its textile dye decolorization potential. LB medium was found to be the most promising growth medium with addition of glucose (1-2%), yeast extract (0.1%), FeCl3 (0.01 mM) and MnCl2 (0.001 mM). The optimum spore laccase production was at pH 8, 30 °C, 1:5 medium to air ratio, 2% inoculum size and 7 days incubation. The characterization study of the enzyme showed the maximum activity at 60 °C and pH 6-7.5. It was induced by Ca+2, Mg+2, Fe+3, Zn+2, Cu+2 and Na+ at 1 mM concentration. Also, it was stable in the presence of methanol, ethanol, acetone and chloroform. In addition, it enhanced about 34% by 5 mM H2O2 and it was nearly stable at 10-20 mM H2O2. Furthermore, mediators such as ABTS, syrengaldazine and 2, 6 dimethyl phenol enhanced the spore laccase activity. The spore laccase enzyme efficiently decolorized direct red 81 and acid black 24 after 24 h. Phytotoxicity of the direct red 81 solution after decolorization by tested spore laccase was lower than that of the untreated dye solution. Finally, this study added a promising spore laccase candidate for ecofriendly and cost-effective dye wastewater bio-decolorization.

Semi-pilot scale production of biodiesel from waste frying oil by genetically improved fungal lipases.

This paper addresses the issue of combining the usage of waste frying oil (WFO), as a feedstock, and a lipase produced in solid-state fermentation (SSF), as a biocatalyst, for semi-pilot scale production of biodiesel as fatty acid methyl esters (FAME). Two fungal mutants namely; Rhizopus stolonifer 1aNRC11 mutant F (1F) and Aspergillus tamarii NDA03a mutant G (3G) were used as a cocatalyst. The two mutants were cultivated separately by SSF in a tray bioreactor. The dried fermented solid of 1F and 3G mutants were used in a ratio of 3:1, respectively, for WFO transesterification. Optimization of several semi-pilot process stages including SSF and WFO transesterification reaction conditions resulted in 92.3% conversion of WFO to FAME. This FAME yield was obtained after 48 h using 10% cocatalyst (w/w of WFO), 10% water (w/w of WFO) and 3:1 methanol/ WFO molar ratio at 30 °C and 250 rpm. A preliminary economic evaluation of produced biodiesel price (190 $/Ton) is less than half the price of petroleum diesel in Egypt (401$/Ton) and is about 40.3% the price of biodiesel produced using a pure enzyme, which is a promising result. This strategy makes the biodiesel synthesis process greener, economical and sustainable.

Multi-bioactive silver nanoparticles synthesized using mosquitocidal Bacilli and their characterization.

Silver nanoparticles (AgNPs) were synthesized using cell-free filtrates of some mosquitocidal Bacilli. They showed the optical absorption peaks at 386-412 nm. They were polycrystalline spherical, hexagonal, cuboidal, rod and anisotropic shapes as detected by TEM. These nanoparticles were negatively charged with sizes ranging from 15 to 21 nm average diameter as detected by DLS. FTIR spectra showed that the main absorption bands of biomolecules capping AgNPs appeared at average wave numbers of 3435 cm-1 [ν(N-H) of amide A overlapped by ν(O-H)], 1631 cm-1 [(ν(C=O) of amide I], 1396 cm-1 [ν(C-N) of amide I], 2929 cm-1 (aliphatic C-H) and 1040 cm-1 (C-C-O). FTIR spectra confirmed the presence of protein biomolecules in the bacterial filtrate-formed coat covering AgNPs through free amide groups resulting in their stabilization in the aqueous medium. Nitrate reductase activity was found in all tested bacterial filtrates and ranged from 1.66 to 2.51 µmol/ml/min. These findings point to the probable role of nitrate reductase in reducing silver ions to silver nanoparticles and their stabilization. Tested AgNPs were multi-bioactive nanometals and showed mosquitocidal, bactericidal, fungicidal and virucidal activities. In addition, they exhibited highly synergistic mosquitocidal effect to spore toxin complex of mosquitocidal Bacilli at a very low concentration. AgNPs exhibited activities that were not or slightly cytotoxic to MA 104 cell line at tested concentrations. Therefore, they can be applied in the medical field. Finally, this study offered a simple, highly efficient, eco-friendly, economic method for biosynthesis of multi-bioactive AgNPs by some mosquitocidal Bacilli.

Efficient mosquitocidal toxin production by Bacillus sphaericus using cheese whey permeate under both submerged and solid state fermentations.

Whey permeate (WP) was used efficiently for production of mosquitocidal toxin by Bacillus sphaericus 2362 (B. sphaericus 2362) and the Egyptian isolate, B. sphaericus 14N1 (B. sphaericus 14N1) under both submerged and solid state fermentation conditions. Under submerged fermentation, high mosquitocidal activity was produced by B. sphaericus 2362 and B. sphaericus 14N1 at 50-100% and 25-70% WP, respectively. Initial pH of WP was a critical factor for toxin production by both tested organisms. The highest toxicity was obtained at initial pH 7. Egyptian isolate, B. sphaericus 14N1 was tested for growth and toxin production under solid state fermentation conditions (SSF) by using WP as moistening agent instead of distilled water. The optimum conditions for production of B. sphaericus 14N1 on wheat bran-WP medium were 10 g wheat bran/250 ml flask moistened with 10-70% WP at 50% moisture content, inoculum size ranged between 17.2x10(7) and 34.4x10(7) and 6 days incubation under static conditions at 30 degrees C. Preliminary pilot-scale production of B. sphaericus 14N1 under SSF conditions in trays proved that wheat bran-WP medium was efficient and economic for industrial production of mosquitocidal toxin by B. sphaericus.

Bacillus thuringiensis and Bacillus sphaericus biopesticides production.

The long residual action and toxicity of the chemical insecticides have brought about serious environmental problems such as the emergence and spread of insecticide resistance in many species of vectors, mammalian toxicity, and accumulation of pesticide residues in the food chain. All these problems have highlighted the need for alternative biological control agents. Entomo-pathogenic Bacillus thuringiensis (Bt) and Bacillus sphaericus (Bs) are two safe biological control agents. They have attracted considerable interest as possible replacements for the chemical insecticides. Although microbial insecticides based on Bt and Bs are available for use, their high cost makes large-scale application impracticable in developing countries. This review focuses on the economic production of these two microorganisms by submerged fermentation and solid state fermentation using agro-industrial by-products and other wastes.