Mohamed El-Agamy Farh

Biosynthesis, characterization, and antimicrobial applications of silver nanoparticles

In the present study, the strain Brevibacterium frigoritolerans DC2 was explored for the efficient and extracellular synthesis of silver nanoparticles. These biosynthesized silver nanoparticles were characterized by ultraviolet-visible spectrophotometry, which detected the formation of silver nanoparticles in the reaction mixture and showed a maximum absorbance at 420 nm. In addition, field emission transmission electron microscopy revealed the spherical shape of the nanoparticles. The dynamic light scattering results indicated the average particle size of the product was 97 nm with a 0.191 polydispersity index. Furthermore, the product was analyzed by energy dispersive X-ray spectroscopy, X-ray diffraction, and elemental mapping, which displayed the presence of elemental silver in the product. Moreover, on a medical platform, the product was checked against pathogenic microorganisms including Vibrio parahaemolyticus, Salmonella enterica, Bacillus anthracis, Bacillus cereus, Escherichia coli, and Candida albicans. The nanoparticles demonstrated antimicrobial activity against all of these pathogenic microorganisms. Additionally, the silver nanoparticles were evaluated for their combined effects with the commercial antibiotics lincomycin, oleandomycin, vancomycin, novobiocin, penicillin G, and rifampicin against these pathogenic microorganisms. These results indicated that the combination of antibiotics with biosynthesized silver nanoparticles enhanced the antimicrobial effects of antibiotics. Therefore, the current study is a demonstration of an efficient biological synthesis of silver nanoparticles by B. frigoritolerans DC2 and its effect on the enhancement of the antmicrobial efficacy of well-known commercial antibiotics.

Biogenic silver and gold nanoparticles synthesized using red ginseng root extract, and their applications

In the present study, we report a green methodology for the synthesis of silver and gold nanoparticles, using the root extract of the herbal medicinal plant Korean red ginseng. The silver and gold nanoparticles were synthesized within 1 h and 10 min respectively. The nanoparticles generated were not aggregated, and remained stable for a long time, which suggests the nature of nanoparticles. The phytochemicals and ginsenosides present in the root extract assist in reducing and stabilizing the synthesized nanoparticles. The red ginseng root extract-generated silver nanoparticles exhibit antimicrobial activity against pathogenic microorganisms including Vibrio parahaemolyticus, Staphylococcus aureus, Bacillus cereus, and Candida albicans. In addition, the silver nanoparticles exhibit biofilm degrading activity against S. aureus and Pseudomonas aeruginosa. Thus, the present study opens up a new possibility of synthesizing silver and gold nanoparticles in a green and rapid manner using Korean red ginseng root extract, and explores their biomedical applications.

Green synthesis of multifunctional silver and gold nanoparticles from the oriental herbal adaptogen: Siberian ginseng.

Pharmacologically active stem of the oriental herbal adaptogen, Siberian ginseng, was employed for the ecofriendly synthesis of Siberian ginseng silver nanoparticles (Sg-AgNPs) and Siberian ginseng gold nanoparticles (Sg-AuNPs). First, for metabolic characterization of the sample, liquid chromatography-tandem mass spectrometry analysis (indicated the presence of eleutherosides A and E), total phenol content, and total reducing sugar were analyzed. Second, the water extract of the sample mediated the biological synthesis of both Sg-AgNPs and Sg-AuNPs that were crystalline face-centered cubical structures with a Z-average hydrodynamic diameter of 126 and 189 nm, respectively. Moreover, Fourier transform infrared analysis indicated that proteins and aromatic hydrocarbons play a key role in the formation and stabilization of Sg-AgNPs, whereas phenolic compounds accounted for the synthesis and stability of Sg-AuNPs. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay determined that Sg-AgNPs conferred strong cytotoxicity against MCF7 (human breast cancer cell line) and was only slightly toxic to HaCaT (human keratinocyte cell line) at 10 µg⋅mL−1. However, Sg-AuNPs did not display cytotoxic effects against both of the cell lines. The disc diffusion assay indicated a dose-dependent increase in the zone of inhibition of Staphylococcus aureus (ATCC 6538), Bacillus anthracis (NCTC 10340), Vibrio parahaemolyticus (ATCC 33844), and Escherichia coli (BL21) treated with Sg-AgNPs, whereas Sg-AuNPs did not show inhibitory activity. In addition, the 2,2-diphenyl-1-picrylhydrazyl assay demonstrated that both Sg-AgNPs and Sg-AuNPs possess strong antioxidant activity. To the best of our knowledge, this is the first report unraveling the potential of Eleutherococcus senticosus for silver and gold nanoparticle synthesis along with its biological applications, which in turn would promote widespread usage of the endemic Siberian ginseng.

In vitro anti-inflammatory activity of spherical silver nanoparticles and monodisperse hexagonal gold nanoparticles by fruit extract of Prunus serrulata: a green synthetic approach

Abstract Recently, green metal nanoparticles have received global attention owing to their economical synthesis, biocompatible nature, widespread biomedical and environmental applications. Current study demonstrates a sustainable approach for the green synthesis of silver nanoparticles (P-AgNPs) and gold nanoparticles (P-AuNPs) from P. serrulata fresh fruit extract. The silver and gold nanoparticles were synthesized in a very rapid, efficient and facile manner, within 50 min and 30 s at 80 °C, respectively. The nanoparticles were characterized by using visual observation, UV–Vis, FE-TEM, EDX, elemental mapping, FT-IR, XRD and DLS, which confirmed the formation of monodispersed, crystalline and stable nanoparticles. Further, we explored these nanoparticles for anti-inflammatory activity through inhibition of downstream NF-κB activation in macrophages (RAW264.7). We demonstrated that the nanoparticles reduced expression of inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PEG2), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was attenuated in lipopolysaccharide (LPS)-induced RAW264.7 cells. Furthermore, nanoparticles significantly suppressed LPS-induced activation of NF-κB signalling pathway via p38 MAPK in RAW 264.7 cells. To the best of our knowledge, this is the first report on the efficient green synthesis of P-AgNPs and P-AuNPs using P. serrulata fresh fruit extract and its in vitro anti-inflammatory effects. Collectively, our results suggest that P. serrulata fresh fruit extract is a green resource for the eco-friendly synthesis of P-AgNPs and P-AuNPs, which further can be utilized as a novel therapeutic agent for prevention and cure of inflammation due to their biocompatible nature.

Cylindrocarpon destructans/Ilyonectria radicicola-species complex: Causative agent of ginseng root-rot disease and rusty symptoms

Cylindrocarpon destructans/Ilyonectria radicicola is thought to cause both rusty symptom and root-rot disease of American and Korean ginseng. Root-rot disease poses a more serious threat to ginseng roots than rusty symptoms, which we argue result from the plant defense response to pathogen attack. Therefore, strains causing rotten root are characterized as more aggressive than strains causing rusty symptoms. In this review, we state 1- the molecular evidence indicating that the root-rot causing strains are genetically distinct considering them as a separate species of Ilyonectria, namely I. mors-panacis and 2- the physiological and biochemical differences between the weakly and highly aggressive species as well as those between rusty and rotten ginseng plants. Eventually, we postulated that rusty symptom occurs on ginseng roots due to incompatible interactions with the weakly aggressive species of Ilyonectria, by the established iron-phenolic compound complexes while root-rot is developed by I. mors-panacis infection due to the production of high quantities of hydrolytic and oxidative fungal enzymes which destroy the plant defensive barriers, in parallel with the pathogen growth stimulation by utilizing the available iron. Furthermore, we highlight future areas for study that will help elucidate the complete mechanism of root-rot disease development.

Cross Interaction Between Ilyonectria mors-panacis Isolates Infecting Korean Ginseng and Ginseng Saponins in Correlation with Their Pathogenicity

Ilyonectria mors-panacis belongs to I. radicicola species complex and causes root rot and replant failure of ginseng in Asia and North America. The aims of this work were to identify I. mors-panacis that infect Korean ginseng using molecular approaches and to investigate whether their aggressiveness depends on their ability to metabolize ginseng saponins (ginsenosides) by their β-glucosidases, in comparison with other identified Ilyonectria species. Fourteen isolates were collected from culture collections or directly isolated from infected roots and mainly identified based on histone H3 (HIS H3) sequence. Among them, six isolates were identified as I. mors-panacis while others were identified as I. robusta and I. leucospermi. The pathogenicity tests confirmed that the isolates of I. mors-panacis were significantly more aggressive than I. robusta and I. leucospermi. The major ginsenosides in I. mors-panacis-infected roots were significantly reduced while significantly increased in those infected with other species. In vitro, the isolates were tested for their sensitivity and ability to metabolize the total major ginsenosides (Total MaG), protopanaxadiol-type major ginsenosides (PPD-type MaG), and protopanaxatriol-type major ginsenosides (PPT-type MaG). Unexpectedly, the growth rate and metabolic ability of I. mors-panacis isolates were significantly low on the three different ginsenoside fractions while those of I. robusta and I. leucospermi were significantly reduced on PPT-type MaG and Total MaG fractions and not affected on PPD-type MaG fraction. Our results indicate that major ginsenosides, especially PPT-type, have an antifungal effect and may intervene in ginseng defense during Ilyonectria species invasion, in particular the weak species. Also, the pathogenicity of I. mors-panacis may rely on its ability to reduce saponin content; however, whether this reduction is caused by detoxification or another method remains unclear.

Phenylobacterium panacis sp. nov., isolated from the rhizosphere of rusty mountain ginseng

A novel, Gram-stain-negative, rod-shaped bacterial strain, designated as DCY109T, was isolated from the rhizosphere of rusty mountain ginseng root located on Hwacheon mountain of Gangwon province, South Korea. 16S rRNA gene sequence analysis revealed that strain DCY109T belonged to the genus Phenylobacterium and was related closely to Phenylobacterium muchangponense KACC 15042T (98.2 % similarity), Phenylobacterium immobile DSM 1986T (96.9 %) and Phenylobacterium koreense KCTC 12206T (96.7 %). The predominant isoprenoid quinone was ubiquinone (Q-10) and the DNA G+C content was 66.9 mol%. The major polar lipids were phosphatidylglycerol, an unidentified glycolipid and an unidentified lipid. The major fatty acids (>10 %) were C16 : 0, summed feature 3 (which comprised C16 : 1ω7c and/or C16 : 1ω6c) and summed feature 8 (which comprised C18 : 1ω7c and/or C18 : 1ω6c). Mean DNA-DNA relatedness between strain DCY109T and its closest relative, P. muchangponense KACC 15042T, was 15.1±3.9 %. Based on the physiological, biochemical, chemotaxonomic and genetic analyses, strain DCY109T is considered to represent a novel species of the genus Phenylobacterium, for which the name Phenylobacterium panacis sp. nov. is proposed. The type strain is DCY109T (=KCTC 42749T=JCM 31045T).

Aluminium resistant, plant growth promoting bacteria induce overexpression of Aluminium stress related genes in Arabidopsis thaliana and increase the ginseng tolerance against Aluminium stress

Panax ginseng is an important cash crop in the Asian countries due to its pharmaceutical effects, however the plant is exposed to various abiotic stresses, lead to reduction of its quality. One of them is the Aluminum (Al) accumulation. Plant growth promoting bacteria which able to tolerate heavy metals has been considered as a new trend for supporting the growth of many crops in heavy metal occupied areas. In this study, twelve bacteria strains were isolated from rhizosphere of diseased Korean ginseng roots located in Gochang province, Republic of Korea and tested for their ability to grow in Al-embedded broth media. Out of them, four strains (Pseudomonas simiae N3, Pseudomonas fragi N8, Chryseobacterium polytrichastri N10, and Burkholderia ginsengiterrae N11-2) were able to grow. The strains could also show other plant growth promoting activities e.g. auxins and siderophores production and phosphate solubilization. P. simiae N3, C. polytrichastri N10, and B. ginsengiterrae N11-2 strains were able to support the growth of Arabidopsis thaliana stressed by Al while P. fragi N8 could not. Plants inoculated with P. simiae N3, C. polytrichastri N10, and B. ginsengiterrae N11-2 showed higher expression level of Al-stress related genes, AtAIP, AtALS3 and AtALMT1, compared to non-bacterized plants. Expression profiles of the genes reveal the induction of external mechanism of Al resistance by P. simiae N3 and B. ginsengiterrae N11-2 and internal mechanism by C. polytrichastri N10. Korean ginseng seedlings treated with these strains showed higher biomass, particularly the foliar part, higher chlorophyll content than non-bacterized Al-stressed seedlings. According to the present results, these strains can be used in the future for the cultivation of ginseng in Al-persisted locations.

Rhodoferax koreense sp. nov, an obligately aerobic bacterium within the family Comamonadaceae, and emended description of the genus Rhodoferax

Gram-staining-negative, uniflagellated, rod-shaped, designated as DCY110T, was isolated from sludge located in Gangwon province, Republic of Korea. The phylogenetic tree of 16S rRNA gene sequence showed that the strain DCY110T belonged to the genus Rhodoferax with a close similarity to Rhodoferax saidenbachensis DSM 22694T (97.7%), Rhodoferax antarcticus DSM 24876T (97.5%), Rhodoferax ferrireducens DSM 15236T (97.3%), and Rhodoferax fermentans JCM 7819T (96.7%). The predominant isoprenoid quinine was ubiquinone (Q-8). DNA G + C content was 62.8 mol%. The major polar lipids were phosphatidylethanolamine and two unidentified phospholipids. The major fatty acids (> 10%) were C12:0, C16:0, summed feature 3 (which comprised C16:1ω7c and/or C16:1ω6c). The DNA-DNA relatedness values between the strain DCY110T and the closely related relatives used in this study were lower than 70%. Based on the following polyphasic analysis, the strain DCY110T is considered as a novel species of the genus Rhodoferax, for which the name Rhodoferax koreense sp. nov. is proposed. The type strain is DCY-110T (= KCTC 52288T = JCM 31441T).

Achromobacter panacis sp. nov., isolated from rhizosphere of Panax ginseng

A novel strain DCY105T was isolated from soil collected from the rhizosphere of ginseng (Panax ginseng), in Gochang, Republic of Korea. Strain DCY105T is Gram-reaction-negative, white, non-motile, non-flagellate, rod-shaped and aerobic. The bacteria grow optimally at 30°C, pH 6.5–7.0 and in the absence of NaCl. Phylogenetically, strain DCY105T is most closely related to Achromobacter marplatensis LMG 26219T (96.81%). The DNA G+C content of strain DCY105T was 64.4 mol%. Ubiquinone 8 was the major respiratory quinone, and phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol were amongst the major polar lipids. C16:00, C8:03OH and iso-C17:03OH were identified as the major fatty acids present in DCY105T. The results of physiological and biochemical tests allowed strain DCY105T to be differentiated phenotypically from other recognized species belonging to the genus Achromobacter. Therefore, it is suggested that the newly isolated organism represents a novel species, for which the name Achromobacter panacis sp. nov. is proposed with the type strain designated as DCY105T (=CCTCCAB 2015193T =KCTC 42751T).