The saponins of
L., saikosaponins, are the major components responsible for its pharmacological and biological activities. However, the anti-cancer effects of prosaikogenin and saikogenin, which are ...glycoside hydrolyzed saikosaponins, are still unknown due to its rarity in plants. In this study, we applied two recombinant glycoside hydrolases that exhibit glycoside cleavage activity with saikosaponins. The two enzymes, BglPm and BglLk, were cloned from
and
, and exhibited good activity between 30-37 °C and pH 6.5-7.0. Saikosaponin A and D were purified and obtained from the crude
L. extract using preparative high performance liquid chromatography technique. Saikosaponin A and D were converted into saikogenin F via prosaikogenin F, and saikogenin G via prosaikogenin G using enzyme transformation with high β-glycosidase activity. The two saikogenin and two prosaikogenin compounds were purified using a silica column to obtain 78.1, 62.4, 8.3, and 7.5 mg of prosaikogenin F, prosaikogenin G, saikogenin F, and saikogenin G, respectively, each with 98% purity. The anti-cancer effect of the six highly purified saikosaponins was investigated in the human colon cancer cell line HCT 116. The results suggested that saikosaponins and prosaikogenins markedly inhibit the growth of the cancer cell line. Thus, this enzymatic technology could significantly improve the production of saponin metabolites of
L.
The ginsenoside compound K (C-K) (which is a de-glycosylated derivative of major ginsenosides) is effective in the treatment of cancer, diabetes, inflammation, allergy, angiogenesis, aging, and has ...neuroprotective, and hepatoprotective than other minor ginsenosides. Thus, a lot of studies have been focused on the conversion of major ginsenosides to minor ginsenosides using glycoside hydrolases but there is no study yet published for the bioconversion of minor ginsenosides into another high pharmacological active compound. Therefore, the objective of this study to identify a new gene (besides the glycoside hydrolases) for the conversion of minor ginsenosides C-K into another highly pharmacological active compound.
which was isolated from Kimchi has showed the ginsenoside C-K altering capabilities. From this strain, a novel potent decarboxylation gene, named HSDLb1, was isolated and expressed in
BL21 (DE3) using the pMAL-c5X vector system. Recombinant HSDLb1 was also characterized. The HSDLb1 consists of 774 bp (258 amino acids residues) with a predicted molecular mass of 28.64 kDa. The optimum enzyme activity was recorded at pH 6.0-8.0 and temperature 30 °C. Recombinant HSDLb1 effectively transformed the ginsenoside C-K to 12-β-hydroxydammar-3-one-20(S)-O-β-D-glucopyranoside (3-oxo-C-K). The experimental data proved that recombinant HSDLb1 strongly ketonized the hydroxyl (-O-H) group at C-3 of C-K via the following pathway: C-K → 3-oxo-C-K. In vitro study, 3-oxo-C-K showed higher solubility than C-K, and no cytotoxicity to fibroblast cells. In addition, 3-oxo-C-K induced the inhibitory activity of ultraviolet A (UVA) against matrix metalloproteinase-1 (MMP-1) and promoted procollagen type I synthesis. Based on these expectations, we hypothesized that 3-oxo-C-K can be used in cosmetic products to block UV radiations and anti-ageing agent. Furthermore, we expect that 3-oxo-C-K will show higher efficacy than C-K for the treatment of cancer, ageing and other related diseases, for which more studies are needed.
It is essential to develop and discover alternative eco-friendly antibacterial agents due to the emergence of multi-drug-resistant microorganisms. In this study, we isolated and characterized a novel ...bacterium named
MAHUQ-38
, utilized for the eco-friendly synthesis of silver nanoparticles (AgNPs) and the synthesized AgNPs were used to control multi-drug-resistant microorganisms. The novel strain was Gram stain positive, strictly aerobic, milky white colored, rod shaped and non-motile. The optimal growth temperature, pH and NaCl concentration were 30 °C, 6.5 and 0%, respectively. Based on 16S rRNA gene sequence, strain MAHUQ-38
belongs to the genus
and is most closely related to several
type strains (98.2%-98.8%).
MAHUQ-38
had a genome of 5,156,829 bp long (19 contigs) with 4555 protein-coding genes, 48 tRNA and 5 rRNA genes. The culture supernatant of strain MAHUQ-38
was used for the eco-friendly and facile synthesis of AgNPs. The transmission electron microscopy (TEM) image showed the spherical shape of AgNPs with a size of 6 to 24 nm, and the Fourier transform infrared (FTIR) analysis revealed the functional groups responsible for the synthesis of AgNPs. The synthesized AgNPs exhibited strong anti-bacterial activity against multi-drug-resistant pathogens,
and
. Minimal inhibitory/bactericidal concentrations against
and
were 6.25/50 and 12.5/50 μg/mL, respectively. The AgNPs altered the cell morphology and damaged the cell membrane of pathogens. This study encourages the use of
for the ecofriendly synthesis of AgNPs to control multi-drug-resistant microorganisms.
Recent years, multi drug resistant pathogens and their pathogenicity were increased worldwide due to unauthorized consumption of antibiotics. In addition, correlation between multi drug resistant ...bacteria and biofilm formation is heightened due to the production of more virulence behavior. There is no better identification methods are available for detection of biofilm producing gram negative bacteria.
In this research work, multi drug resistant strains of Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae) were identified based on the specific antibiotics and third generation cephalosporin discs by disc diffusion assay. Subsequently, biofilm forming ability of selected pathogens were identified tissue culture plate and tube test. Based on the multi-drug resistant ability and biofilm production, the molecular identification of P. aeruginosa and K. pneumoniae were confirmed by PCR using universal primers.
No zone of inhibition present around the discs of muller hinton agar plates were confirm, selected P. aeruginosa and K. pneumoniae strains were multi drug resistant pathogens. Performed third generation cephalosporin antibiotics were also highly sensitive to selected pathogens of P. aeruginosa and K. pneumoniae. Further, biofilm forming ability of selected P. aeruginosa and K. pneumoniae was confirmed by tissue culture plate and tube methods. Finally, molecular identification of P. aeruginosa and K. pneumoniae was named as P. aeruginosa and K. pneumoniae. Our result was conclude, selected P. aeruginosa and K. pneumoniae as biofilm producing pathogens and also highly resistant to current antibiotics.
In this study, red ginseng extract (RGE) was converted into high-content minor ginsenosides by fermenting with Bgp1 enzymes at 37°C for 5 days. Compared to the RGE, the minor ginsenoside contents ...were increased in fermented red ginseng extract (FRGE). Moreover, the amount of minor ginsenosides such as Rh1 (11%) and Rg2 (16%) was slightly augmented, while the level of Rg3 (33%) was significantly increased after bioconversion. Furthermore, we also examined and compared the effect of RGE and FRGE on the differentiation and mineralization of preosteoblastic MC3T3-E1 cells. Similarly, the level of mRNA expression of intracellular alkaline phosphatase (ALP) activity, type-1 collagen (Col-I) was also increased. Based on the comparison, it is clear that the FRGE has improved effects on bone formation and differentiation of preosteoblastic MC3T3-E1 cells.
Worldwide, multi-drug resistant Klebsiella pneumoniae (K. pneumonia) and their virulence’s were contributed more in the multi-drug resistant effect. According to the World Health organization report, ...it is an emerging thread in developing countries and also comes under first ever critical list. In this context, the current study was concentrated on detection of extended spectrum beta lactamase (ESBL) producing strain and their antimicrobial susceptibility study of K. pneumoniae.
Firstly, the multi-drug resistant effect of the K. pneumoniae was identified from specific CLSI guidelines recommended antibiotics by disc diffusion method. Consecutively, the primary ESBL identification test was performed using ceftazidime and cefotaxime, followed by double disc combination and phenotypic confirmation tests using ceftazidime/clavulanic acid and cefotaxime/clavulanic acid. Finally, the minimum inhibition concentration of some important sensitive antibiotics were performed against selected K. pneumoniae was confirmed by micro broth dilution method.
The current result was most favorable to selected K. pneumoniae with more multi drug resistant characteristic nature. All the performed antibiotics were almost more sensitive to selected K. pneumoniae. The effective antibiotics of piperacillin was also exhibited more resistant effect against tested bacteria and it cleaved the bacterial enzyme clearly. The present result of primary ESBL identification test result was exhibited with ≤22 mm and ≤27 mm against ceftazidime and cefotaxime were observed respectively. Followed result of double disc combination and phenotypic confirmation experiments results were clearly stated that the selected K. pneumoniae was ESBL producer. The ceftazidime, cefotaxime and ceftazidime/clavulanic acid and cefotaxime/clavulanic acid were exhibited with merged zones and ≥5 mm zones around the combination disc when compared with disc alone were observed. All the ESBL detection test results were clearly indicated that the selected K. pneumoniae strain was ESBL producer.
Several studies have reported that ginsenoside Rg3(
) is effective in treating metastatic diseases, obesity, and various cancers, however, its presence in white ginseng cannot be estimated, and only ...a limited amount is present in red ginseng. Therefore, the use of recombinant glycosidases from a Generally Recognized As Safe (GRAS) host strain is a promising approach to enhance production of Rg3(
), which may improve nutritional activity, human health, and quality of life.
EMML 3041
, which was isolated from Korean fermented pickle (kimchi), presents ginsenoside-converting abilities. The strain was used to enrich the production of Rg3(
) by fermenting protopanaxadiol (PPD)-mix-type major ginsenosides (Rb1, Rb2, Rc, and Rd) in four different types of food-grade media (1, MRS; 2, Basel Food-Grade medium; 3, Basel Food-Grade medium-I, and 4, Basel Food-Grade medium-II). Due to its tendency to produce Rg3(
), the presence of glycoside hydrolase in
was proposed, the whole genome was sequenced, and the probable glycoside hydrolase gene for ginsenoside conversion was cloned.
The
EMML 3041
strain was whole genome sequenced to identify the target genes. After genome sequencing, 12 sets of glycoside hydrolases were identified, of which seven sets (α,β-glucosidase and α,β-galactosidase) were cloned in
BL21 (DE3) using the pGEX4T-1 vector system. Among the sets of clones, only one clone (BglL.gin-952) showed ginsenoside-transforming abilities. The recombinant BglL.gin-952 comprised 952 amino acid residues and belonged to glycoside hydrolase family 3. The enzyme exhibited optimal activity at 55 °C and a pH of 7.5 and showed a promising conversion ability of major ginsenoside Rb1→Rd→Rg3(
). The recombinant enzyme (GST-BglL.gin-952) was used to mass produce Rg3(
) from major ginsenoside Rb1. Scale-up of production using 50 g of Rb1 resulted in 30 g of Rg3(
) with 74.3% chromatography purity.
Our preliminary data demonstrated that this enzyme would be beneficial in the preparation of pharmacologically active minor ginsenoside Rg3(
) in the functional food and pharmaceutical industries.
Initially, the excellent anti-oxidant property of the Ag NPs was synthesized from Chinese medicinal plant of Lonicera japonica, and their spectroscopic evidences were confirmed by UV-spectrometer. ...The morphology, size and shape of the synthesized Ag NPs were effectively identified by scanning electron microscope and transmission electron microscope. Both the spectroscopic results were more evident, which UV-spectrometer peak was exhibited at 456 nm, scanning electron microscope and transmission electron microscope results were also clearly shown ball like spherical shape. The potential anti-cancer effect of the Ag NPs was revealed, the selected Chinese plant Lonicera japonica was influenced the A549 lung cancer cells through excessive ROS production. The IC50 concentration of 75 µg/mL was shown decreased viability in Ag NPs treated A549 lung cancer cells of MTT assay. Consecutively, the MTT assay result was proved by irregular shape and damaged morphology in A549 lung cancer cells by images of phase contrast microscope. Finally, the result was also clearly stated that the synthesized Ag NPs was very effective against A549 human lung cancer cells and it use for future drug discovery for inhibiting the lung cancer.
Background
Rg3-ginsenoside, a protopanaxadiol saponin, is a well-known adaptogen used for the prevention of cancer and inflammation. However, despite its distinct biological activity, the ...concentration of Rg3 in the total ginseng extract is insufficient for therapeutic applications. This study aims to convert PPD-class of major ginsenosides into a mixture of minor ginsenoside, to analyze its immune-regulatory role in macrophage cells.
Results
Using heat and organic acid treatment, three major ginsenosides, Rc, Rd, and Rb1, were converted into a mixture of minor ginsenosides, GRg3-mix Rg3(
S
), Rg3(
R
), Rg5, and Rk1. Purity and content analysis of the transformed compound were performed using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC), compared with their standards. Preceding with the anti-inflammatory activity of GRg3-mix, lipopolysaccharide (LPS)-stimulated murine RAW264.7 macrophage cells were treated with various concentrations of GRg3-mix (6.25, 12.5, 25, and 50 μg/mL). The cell viability assay revealed that the level of cell proliferation was increased, while the nitric oxide (NO) assay showed that NO production decreased dose-dependently in activated RAW264.7 cells. The obtained results were compared to those of pure Rg3(
S
) ≥ 98% (6.25, 12.5, and 25 μg/mL). Preliminary analysis of the CCK-8 and NO assay demonstrated that GRg3-mix can be used as an anti-inflammatory mediator, but mRNA and protein expression levels were evaluated for further confirmation. The doses of GRg3-mix significantly suppressed the initially upregulated mRNA and protein expression of inflammation-related enzymes and cytokines, namely inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear transcription factor kappa B (NF-κB), tumor necrosis factor (TNF-α), and interleukins (IL-6 and IL1B), as measured by reverse transcription-polymerase chain reaction and western blotting.
Conclusions
Our pilot data confirmed that the mixture of minor ginsenosides, namely GRg3-mix, has high anti-inflammatory activity and has an easy production procedure.
The ginseng plant (Panax ginseng Meyer) has a large number of active ingredients including steroidal saponins with a dammarane skeleton as well as protopanaxadiol and protopanaxatriol, commonly known ...as ginsenosides, which have antioxidant, anticancer, antidiabetic, anti-adipocyte, and sexual enhancing effects. Though several discoveries have demonstrated that ginseng saponins (ginsenosides) as the most important therapeutic agent for the treatment of osteoporosis, yet the molecular mechanism of its active metabolites is unknown. In this review, we summarize the evidence supporting the therapeutic properties of ginsenosides both in vivo and in vitro, with an emphasis on the different molecular agents comprising receptor activator of nuclear factor kappa-B ligand, receptor activator of nuclear factor kappa-B, and matrix metallopeptidase-9, as well as the bone morphogenetic protein-2 and Smad signaling pathways.
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