RESUMO
Ganoderma species have been used in folk medicine against different illnesses and are characterized by producing a diversity of bioactive metabolites (triterpenoids, polysaccharides, flavonoids, and phenols) with numerous medicinal effects (anti-proliferative, antioxidant, anti-inflammatory, and antibacterial). This work aims to evaluate ethanolic extracts of fruiting bodies of Ganoderma oerstedii, G. weberianum, and G. subincrustatum strains from the Sonoran Desert in the anti-proliferative activity by the MTT assay on cancer cell lines; anti-inflammatory effect by quantifying nitric oxide (NO) production; antioxidant activity by DPPH, ABTS, and FRAP assays; total phenolic and flavonoid content by Folin-Ciocalteu and AlCl3 method, respectively; antibacterial activity by the broth microdilution method against Escherichia coli and Staphylococcus aureus. Extracts showed anti-proliferative activity with IC50 < 100 µg/mL on the cancer cell lines MDA-MB-231, A549, and HeLa, except G. subincrustatum extract with an IC50 > 100 µg/mL; anti-proliferative activity was not selective, being affected non-cancerous cell line ARPE-19. Extracts showed significant inhibition of NO release in cells stimulated by LPS, up to 60% with G. subincrustatum and G. oerstedii, and 47% with G. weberianum. All tested assays showed moderate antioxidant potential; the most active was G. lucium (control strain) with IC50 of 69 and 30 µg/mL by DPPH and ABTS respectively; and 271 µg Trolox equivalents/g by FRAP. Total phenols and flavonoids ranged from 38 to 56 mg GAE/g and 0.53 to 0.93 mg QE/g, respectively. A significant correlation was found between the antioxidant activities revealed by DPPH, ABTS, and FRAP with total phenol and flavonoid contents. Antibacterial activity was weak against S. aureus (MIC50 > 10 mg/mL). These results demonstrate that tested Ganoderma mushrooms have medicinal potential such as anti-inflammatory and anti-proliferative.
Assuntos
Antioxidantes , Ganoderma , Antioxidantes/farmacologia , Antioxidantes/química , México , Staphylococcus aureus , Extratos Vegetais/química , Antibacterianos/farmacologia , Fenóis/análise , Ganoderma/química , Flavonoides/farmacologia , Anti-Inflamatórios/farmacologiaRESUMO
Recently, several studies have been published on the application of plasmid DNA (pDNA) in gene therapy and vaccine production. The bioprocess to obtain pDNA involves the steps of fermentation, primary recovery, secondary recovery and final purification. The pDNA primary recovery, which is the key step to the rest of the process, includes biomass separation, alkaline lysis and clarification of the lysate. In this work, the clarification by depth bed microfiltration of lysates of E. coli DH5α containing the plasmid pVAX1-NH36 was investigated. The studies were conducted using filter capsules with nominal 8.0 µm pore size using fluxes of 0.0027 and 0.004 cm(3)/(cm(2)-s). The results were compared with the conventional clarification by centrifugation. A fiber coating model was used to describe the behavior of the microfiltration system. A 99 % of solids elimination of the lysates was achieved with depth bed filtration method. The removed solids occupied 23 and 43 % (for 4 and 6 cm(3)/min, respectively) of the void volume of the depth bed microfiltration capsule since an early breakthrough curve is characteristic of these processes. The depth bed microfiltration process for removal of solids from the cell lysate showed competitive results compared to clarification by centrifugation.
Assuntos
DNA/isolamento & purificação , Escherichia coli/genética , Filtração/métodos , Plasmídeos , Cromatografia Líquida de Alta Pressão , Meios de Cultura , Eletroforese em Gel de ÁgarRESUMO
A new bioprocess using mainly membrane operations to obtain purified plasmid DNA from Escherechia coli ferments was developed. The intermediate recovery and purification of the plasmid DNA in cell lysate was conducted using hollow-fiber tangential filtration and tandem anion-exchange membrane chromatography. The purity of the solutions of plasmid DNA obtained during each process stage was investigated. The results show that more than 97% of RNA in the lysate was removed during the process operations and that the plasmid DNA solution purity increased 28-fold. One of the main characteristics of the developed process is to avoid the use of large quantities of precipitating agents such as salts or alcohols. A better understanding of membrane-based technology for the purification of plasmid DNA from clarified E. coli lysate was developed in this research. The convenience of anion-exchange membranes, configured in ready-to-use devices can further simplify large-scale plasmid purification strategies.