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- A bi-layer electrospun nanofiber membrane for plasmid DNA recovery from fermentation brothsPublication . Correia, Tiago R.; Antunes, Bernardo Paiva; Castilho, Pedro Henrique Barata; Nunes, José; Amorim, Maria T. Pessoa de; Escobar, Isabel; Queiroz, João; Correia, I.J.; Morão, AntónioThe demanding ever-increasing quantities of highly purified biomolecules by bio-industries, has triggered the development of new, more efficient, purification techniques. The application of membrane-based technologies has become very attractive in this field, for their high throughput capability, simplicity of operation and scale-up. Herein we report the production of a bi-layer membrane by electrospinning (ES), in which a support of poly ε-caprolactone nanofibers was coated with a polyethylene oxide/sodium alginate layer, and subsequently cross-linked with calcium chloride. The membranes were characterized by SEM, ATR-FTIR, contact angle measurements, and were applied in the recovery process of a plasmid. The results show that membranes retained the suspended solids while allowing the permeation of plasmid DNA, with high recovery yields and improved RNA retention. Moreover, they also showed a very low fouling tendency. To the best of our knowledge it is the first time that ES membranes are applied in this type of bioprocess.
- Biological treatment of effluent containing textile dyesPublication . Gonçalves, Isolina Maria da Silva Cabral; Amorim, M. T. P.; Porter, R. S.; Gomes, Arlindo Caniço; Ferra, Maria Isabel; Brás, RuiColour removal of textile dyes from effluent was evaluated using a laboratory upflow anaerobic sludge blanket reactor. Several commercial dyes were selected to study the effect of dye structure on colour removal. The anaerobic reactor was fed with glucose, an easily biodegradable organic matter and selected individual dyes. Results show that some of the dyes are readily reduced under anaerobic conditions even at high concentration of 700 mg/l. The average removal efficiency for acid dyes using this method was between 80 and 90% and that observed for the direct used was 81%. Laboratory experiments using the anaerobic reactor with disperse dyes, such as an anthraquinone based dye, were unsuccessful even at low concentrations of 35 mg/l. Additional experiments were conducted to evaluate the toxicity of a selected disperse dye to an anaerobic environment. Results indicate that the purified dye is more toxic to the biomass than the commercial one.
- Modification of microfiltration membranes by hydrogel impregnation for pDNA purificationPublication . Castilho, Pedro Henrique Barata; Correia, Tiago R.; Amorim, Maria T. Pessoa de; Escobar, Isabel; Queiroz, João; Correia, Ilídio Joaquim Sobreira; Morão, AntónioThe huge efforts for the achievement of highly purified biomolecules are growing every day. A great number of efficient techniques, such as chromatography, are already available in laboratory for separation processes. However, membrane‐based technologies are the best match to assure simplicity, efficiency and easy scale‐up procedures. Herein we report the modification of a commercial microfiltration membrane for plasmid DNA purification by agarose gel impregnation. The membrane was characterized by SEM, ATR‐FTIR, EDS, contact angle, and porosity measurements. Additionally, the membrane pore radius was estimated from observed rejections of different proteins and with that information the rejection of a 6050 bp plasmid DNA (pDNA) molecule was estimated for different values of flux using a theoretical model of large flexible molecules in membranes with parallel cylindrical pores, which is applicable to pDNA ultrafiltration in conventional membranes, as recently shown in the literature. The experimental results show that the modified membrane has higher pDNA rejections than the predicted by the model, suggesting that the different type of porous structure that a hydrogel has, may have a positive effect on pDNA rejections as compared to other biomolecules with more rigid structures, making this type of modified membranes potential better candidates to be used for the selective recovery of pDNA in this type of bioprocesses.