Separation Properties of Plasmid DNA Using a Two-Stage Particle Adsorption-Microfiltration Process.

Plasmid DNA is used as a vector for gene therapy and DNA vaccination; therefore, the establishment of a mass production method is required. Membrane filtration is widely employed as a separation method suitable for the mass production of plasmid DNA. Furthermore, the separation of plasmid DNA using microfiltration and ultrafiltration membranes is being investigated. Because plasmid DNA has a circular structure, it undergoes significant deformation during filtration and easily permeates the membrane, hindering the selection of separation membranes based on molecular weight. In this study, we applied affinity microfiltration to plasmid DNA purification. α-Fe2O3 with an isoelectric point of approximately 8 and a particle size of 0.5 µm was selected as the ligand for two-stage affinity microfiltration of plasmid DNA. In the first stage of microfiltration, the experiment was conducted at a pH of 5, and a cake of α-Fe2O3 with bound plasmid DNA was obtained. Next, liquid permeation (pH 9 and 10) through the cake was performed to elute the bound plasmid DNA. Plasmid DNA was eluted during the early phase of liquid permeation at pH 10. Furthermore, agarose gel analysis confirmed the usefulness of the two-stage affinity microfiltration method with adsorption and desorption for plasmid DNA purification.

Recovery of Extracellular Polymeric Substances from Excess Sludge Using High-Flux Electrospun Nanofiber Membranes.

The recycling of extracellular polymeric substances (EPSs) from excess sludge in wastewater treatment plants has received increasing attention in recent years. Although membrane separation has great potential for use in EPS concentration and recovery, conventional membranes tend to exhibit low water flux and high energy consumption. Herein, electrospun nanofiber membranes (ENMs) were fabricated using polyvinylidene fluoride (PVDF) and used for the recovery of EPSs extracted from the excess sludge using the cation exchange resin (CER) method. The fabricated ENM containing 14 wt.% PVDF showed excellent properties, with a high average water flux (376.8 L/(m2·h)) and an excellent EPS recovery rate (94.1%) in the dead-end filtration of a 1.0 g/L EPS solution at 20 kPa. The ENMs displayed excellent mechanical strength, antifouling properties, and high reusability after five recycles. The filtration pressure had a negligible effect on the average EPS recovery rate and water flux. The novel dead-end filtration with an EPS filter cake on the ENM surface was effective in removing heavy-metal ions, with the removal rates of Pb2+, Cu2+, and Cr6+ being 89.5%, 73.5%, and 74.6%, respectively. These results indicate the potential of nanofiber membranes for use in effective concentration and recycling of EPSs via membrane separation.

Yeast Cell Cake Characterization in Alcohol Solution for Efficient Microfiltration.

Microfiltration is widely used to remove microbial cells from the fermentation broth in the downstream processing of biotechnological products. Because filtration behaviors are strongly affected by the characteristics of the microbial cell cake formed on the surface of the membrane, insights into the cake structure facilitate the design and operation of filter equipment and membranes. In the alcohol fermentation process using a yeast strain, the cake characteristics are considered to be complicated because yeast cells are strongly influenced by external factors such as filtration pressure and alcohol concentration. In this study, we evaluated the membrane filtration properties, in particular the cake characteristics of a yeast suspension containing alcohol. Microfiltration experiments were performed in the dead-end filtration mode using yeast suspensions with several ethanol concentrations (0-20 wt%) under constant pressure. Flux decline behaviors caused by yeast cake were put in a similar form for 0-15 wt% ethanol concentrations. In contrast, a severe flux decline was observed for the suspension with 20 wt% ethanol concentration. It was also observed that in the membrane filtration of yeast cells with 20 wt% ethanol concentration, the cake structure became denser and the filtration resistance remarkably increased because of cellular destruction. Furthermore, the yeast cake exhibited a high compressibility in the solution containing a 20 wt% ethanol concentration. Therefore, the filtration rate of the alcoholic fermentation broth is not significantly improved by increased pressure due to the increase in the cake resistance.