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This work aims at understanding the attachment mechanisms and stability of proteins on a chromatography medium to develop more efficient functionalization methodologies, which can be exploited in affinity chromatography. In particular, the study was focused on the understanding of the attachment mechanisms of bovine serum albumin (BSA), used as a ligand model, and protein G on novel amine-modified alumina monoliths as a stationary phase. Protein G was used to develop a column for antibody purification. The results showed that, at lower protein concentrations (i.e., 0.5 to 1.0 mg·mL−1), protein attachment follows a 1st-order kinetics compatible with the presence of covalent binding between the monolith and the protein. At higher protein concentrations (i.e., up to 10 mg·mL−1), the data preferably fit a 2nd-order kinetics. Such a change reflects a different mechanism in the protein attachment which, at higher concentrations, seems to be governed by physical adsorption resulting in a multilayered protein formation, due to the presence of ligand aggregates. The threshold condition for the prevalence of physical adsorption of BSA was found at a concentration higher than 1.0 mg·mL−1. Based on this result, protein concentrations of 0.7 and 1.0 mg·mL−1 were used for the functionalization of monoliths with protein G, allowing a maximum attachment of 1.43 mg of protein G/g of monolith. This column was then used for IgG binding–elution experiments, which resulted in an antibody attachment of 73.5% and, subsequently, elution of 86%, in acidic conditions. This proved the potential of the amine-functionalized monoliths for application in affinity chromatography. [ABSTRACT FROM AUTHOR]

Separation and high recovery factor of proteins similar in molecular mass is a challenging task, and heavily studied in the literature. In this work, a systematic study to separate a binary protein mixture by charged ultrafiltration membranes without affecting membrane performance was carried out. α-lactalbumin (ALA, 14.4 kDa) and β-lactoglobulin (BLG, 18.4 kDa) were used as a binary model system. These two proteins are the main proteins of whey, a very well-known byproduct from the dairy industry. Initially, a systematic characterization of individual proteins was carried out to determine parameters (protein size and aggregation, zeta potential) which could influence their passage through a charged membrane. Then, the influence of operating parameters (such as initial protein concentration, pH, and critical pressure) on the UF process was investigated, so as to identify conditions that limit membrane fouling whilst maximizing protein recovery factor and purity. The study permitted to identify process conditions able to fully separate ALA from BLG, with high purity (95%) and recovery factor (80%), in a single UF step. Compared to studies reported in literature, here, the main approach used was to carry out a charged UF process far from proteins isoelectric point (pI) to limit protein aggregation and membrane fouling. [ABSTRACT FROM AUTHOR]

Featured Application: Investigation and prediction of crystallization and nucleation of mono and bivalent crystals with a new class of materials in advanced membrane separations. Membrane-assisted crystallization, aiming to induce supersaturation in a solution, has been successfully tested in the crystallization of ionic salts, low molecular organic acids, and proteins. Membrane crystallization is an emerging membrane process with the capability to simultaneously extract fresh water and valuable components from various streams. Successful application of crystallization for produced water treatment, seawater desalination, and salt recovery has been demonstrated. Recently, membrane crystallization has been developed to recover valuable minerals from highly concentrated solutions, since the recovery of high-quality minerals is expected to impact agriculture, pharmaceuticals, and household activities. In this work, molecular dynamics simulations were used to study the crystal nucleation and growth of sodium chloride in bulk and with hydrophobic polymer surfaces of polyvinylidene fluoride (PVDF) and polypropylene (PP) at a supersaturated concentration of salt. In parallel, membrane crystallization experiments were performed utilizing the same polymeric membranes in order to compare the experimental results with the computational ones. Moreover, the comparison in terms of nucleation time between the crystallization of sodium chloride (NaCl) using the traditional evaporation process and the membrane-assisted crystallization process was performed. Here, with an integrated experimental–computational approach, we demonstrate that the PVDF and PP membranes assist the crystal growth for NaCl, speeding up crystal nucleation in comparison to the bulk solution and leading to smaller and regularly structured face-centered cubic lattice NaCl crystals. This results in a mutual validation between theoretical data and experimental findings and provides the stimuli to investigate other mono and bivalent crystals with a new class of materials in advanced membrane separations. [ABSTRACT FROM AUTHOR]

A photoactive gel has been fabricated on the surface of polyethylene membranes for enhancing the fouling resistance during olive mill wastewater treatment. Light and pH responsive materials have been introduced in the membrane surface through the build up of a layer-by-layer pattern, which is formed by photocatalytic nanoparticles and ionic polyelectrolytes. The best working conditions to contrast foulants adsorption have been explored and identified. Repulsive interfacial forces and assisted transfer of foulants to catalytic sites have been envisaged as crucial factors for contrasting the decline of the flux during microfiltration. Tests in submerged configuration have been implemented for six continuous hours under irradiation at two different pH conditions. As a result, a worthy efficiency of the photoactive gel has been reached when suitable chemical microenvironments have been generated along the shell side of the membranes. No additional chemical reagents or expensive back-flushing procedures have been necessary to further clean the membranes; rather, fast and reversible pH switches have been enough to remove residues, thereby preserving the integrity of the layer-by-layer (LBL) complex onto the membrane surface. [ABSTRACT FROM AUTHOR]

Biophenols are highly prized for their free radical scavenging and antioxidant activities. Olive mill wastewaters (OMWWs) are rich in biophenols. For this reason, there is a growing interest in the recovery and valorization of these compounds. Applications for the encapsulation have increased in the food industry as well as the pharmaceutical and cosmetic fields, among others. Advancements in micro-fabrication methods are needed to design new functional particles with target properties in terms of size, size distribution, and functional activity. This paper describes the use of the membrane emulsification method for the fine-tuning of microparticle production with biofunctional activity. In particular, in this pioneering work, membrane emulsification has been used as an advanced method for biophenols encapsulation. Catechol has been used as a biophenol model, while a biophenols mixture recovered from OMWWs were used as a real matrix. Water-in-oil emulsions with droplet sizes approximately 2.3 times the membrane pore diameter, a distribution span of 0.33, and high encapsulation efficiency (98% ± 1% and 92% ± 3%, for catechol and biophenols, respectively) were produced. The release of biophenols was also investigated. [ABSTRACT FROM AUTHOR]

Biocatalytic membrane reactors (BMR) combining reaction and separation within the same unit have many advantages over conventional reactor designs. Ceramic membranes are an attractive alternative to polymeric membranes in membrane biotechnology due to their high chemical, thermal and mechanical resistance. Another important use is their potential application in a biphasic membrane system, where support solvent resistance is highly needed. In this work, the preparation of asymmetric ceramic hollow fibre membranes and their use in a two-separate-phase biocatalytic membrane reactor will be described. The asymmetric ceramic hollow fibre membranes were prepared using a combined phase inversion and sintering technique. The prepared fibres were then used as support for lipase covalent immobilization in order to develop a two-separate-phase biocatalytic membrane reactor. A functionalization method was proposed in order to increase the density of the reactive hydroxyl groups on the surface of ceramic membranes, which were then amino-activated and treated with a crosslinker. The performance and the stability of the immobilized lipase were investigated as a function of the amount of the immobilized biocatalytst. Results showed that it is possible to immobilize lipase on a ceramic membrane without altering its catalytic performance (initial residual specific activity 93%), which remains constant after 6 reaction cycles. [ABSTRACT FROM AUTHOR]

The agri-food sector generates substantial quantities of waste material on farm and during the processing of these commodities, creating serious social and environmental problems. However, these wastes can be resources of raw material for the production of valuable chemicals with applications in various industrial sectors (e.g., food ingredients, nutraceuticals, bioderived fine chemicals, biofuels etc.). The recovery, purification and biotransformation of agri-food waste phytochemicals from this microbial spoilage-prone, complex agri-food waste material, requires appropriate fast pre-treatment and integration of various processes. This review provides a brief summary and discussion of the unique advantages and the importance of membrane technology in sustainable recycling of phytochemicals from some of the main agri-food sectors. Membrane-based pressure -driven processes present several advantages for the recovery of labile compounds from dilute streams. For example, they are clean technologies that can operate at low temperature (20–60 °C), have low energy requirements, there is no need for additional chemicals, can be quite automated and electrifiable, and have low space requirements. Based on their permselective properties based on size-, shape-, and charge-exclusion mechanisms, membrane-based separation processes have unpaired efficiency in fractionating biological components while presenting their properties. Pressure-driven membrane processes, such as microfiltration (MF), ultrafiltration (UF) and nanofiltration (NF), as well as other advanced membrane-based processes such as membrane bioreactors (MBR), membrane emulsification (ME) and membrane distillation (MD), are presented. The integration of various membrane technologies from the initial recovery of these phytochemicals (MF, UF, NF) to the final formulation (by ME) of commercial products is described. A good example of an extensively studied agri-food stream is the olive processing industry, where many different alternatives have been suggested for the recovery of biophenols and final product fabrication. Membrane process integration will deliver in the near future mature technologies for the efficient treatment of these streams in larger scales, with direct impact on the environmental protection and society (production of compounds with positive health effects, new job creation, etc.). It is expected that integration of these technologies will have substantial impact on future bio-based societies over forthcoming decades and change the way that these chemicals are currently produced, moving from petrochemical-based linear product fabrication to a sustainable circular product design based in agri-food waste biomass. [ABSTRACT FROM AUTHOR]

Multiphase bioreactors using interfacial biocatalysts are unique tools in life sciences such as pharmaceutical and biotechnology. In such systems, the formation of microdroplets promotes the mass transfer of reagents between two different phases, and the reaction occurs at the liquid–liquid interface. Membrane emulsification is a technique with unique properties in terms of precise manufacturing of emulsion droplets in mild operative conditions suitable to preserve the stability of bioactive labile components. In the present work, membrane emulsification technology was used for the production of a microstructured emulsion bioreactor using lipase as a catalyst and as a surfactant at the same time. An emulsion bioreaction system was also prepared by the stirring method. The kinetic resolution of (S,R)-naproxen methyl ester catalyzed by the lipase from Candida rugosa to obtain (S)-naproxen acid was used as a model reaction. The catalytic performance of the enzyme in the emulsion systems formulated with the two methods was evaluated in a stirred tank reactor and compared. Lipase showed maximum enantioselectivity (100%) and conversion in the hydrolysis of (S)-naproxen methyl ester when the membrane emulsification technique was used for biocatalytic microdroplets production. Moreover, the controlled formulation of uniform and stable droplets permitted the evaluation of lipase amount distributed at the interface and therefore the evaluation of enzyme specific activity as well as the estimation of the hydrodynamic radius of the enzyme at the oil/water (o/w) interface in its maximum enantioselectivity. [ABSTRACT FROM AUTHOR]