Drop formation in aqueous two-phase systems.

Extraction using aqueous two-phase systems (ATPSs) is a versatile technique for the downstream processing of various proteins/enzymes. The study of drop formation deals with the fundamental understanding of the behavior of liquid drops under the influence of various external body as well as surface forces. These studies provide a basis for designing of the extractions in column contactors in which liquid drops play a major role. Most of the drop formation studies reported so far is restricted to aqueous-organic systems. ATPSs, differ from aqueous-organic systems in their physical properties. In view of this, an attempt was made to develop a model for drop formation in ATPSs adopting the information available on aqueous-organic systems. In order to validate the model, experiments were performed by using polyethylene glycol (PEG)/salt systems of different phase compositions at various flow rates. At low flow rates the single stage model and at high flow rates the two stage model are able to predict the drop volume during its formation from tip of capillary. The experimental results were found to agree reasonably well with those predicted by the model.

Microwave-field-assisted enhanced demixing of aqueous two-phase systems.

The slow rate of demixing is a major limitation in wide commercial exploitation of aqueous two-phase systems. In the present work, use of a microwave field has been explored for the first time to enhance phase demixing rates (decrease demixing times) of these systems. The microwave-field-assisted demixing process decreased the demixing time by about 2- to 4-fold in a polyethylene glycol/potassium phosphate system and by about 1.5- to 6.5-fold in a polyethylene glycol/maltodextrin system. The enhanced demixing rate can be explained by the dipole rotation, electrophoretic migration of free salts, multiple reflections at the interfaces, droplet-droplet collision, and reduced viscosity of the continuous phase that occur during the application of a microwave field.

Acoustic field assisted demixing of aqueous two-phase systems.

Acoustic field assisted demixing was employed to decrease the demixing time in aqueous two-phase systems (polyethylene glycol-maltodextrin and polyethylene glycol-potassium phosphate). Application of acoustic field has decreased the demixing time in polyethylene glycol-maltodextrin by around twofold and up to about 3.2-fold in polyethylene glycol-potassium phosphate systems. Ultrasonication has induced mild circulation currents in the phase dispersion, which has enhanced the rate of droplet coalescence, eventually resulting in decreased demixing time. In the polyethylene glycol-maltodextrin system, phase demixing was found to depend greatly on which of the phases iscontinuous and viscosity of the continuous phase was observed to have a strong influence on the movement of the droplets and hence controlling the phase demixing rate. In case of the polyethylene glycol-potassium phosphate system, droplet coalescence was found to play a critical role in phase demixing. Addition of NaCl increased the demixing time and presence of Escherichia coli cells did not seem to have any influence on phase demixing.

Reverse micellar extraction for downstream processing of proteins/enzymes.

New developments in the area of downstream processing are, hopefully, to fulfill the promises of modern biotechnology. The traditional separation processes such as chromatography or electrophoresis can become prohibitively expensive unless the product is of high value. Hence, there is a need to develop efficient and cost-effective downstream processing methods. Reverse micellar extraction is one such potential and a promising liquid-liquid extraction technique, which has received immense attention for isolation and purification of proteins/enzymes in the recent times. This technique is easy to scale-up and offers continuous operation. This review, besides briefly considering important physico-chemical and biological aspects, highlights the engineering aspects including mass transfer, mathematical modeling, and technology development. It also discusses recent developments in reverse micellar extraction such as affinity based separations, enzymatic reactions in reverse micelles coupled with membrane processes, reverse micellar extraction in hollow fibers, etc. Special emphasis has been given to some recent applications of this technique.

Acoustic field assisted enhanced demixing of aqueous two-phase systems.

Aqueous two-phase extraction has been recognized as a versatile downstream processing technique for the recovery of biomolecules. A major deterrent to its industrial exploitation is the slow demixing of the two aqueous phases after extraction, due to their similar physical properties. A method to decrease the demixing times of these systems, employing a travelling acoustic wave field, is reported. The effects of phase composition and microbial cells on demixing in a polyethylene glycol/potassium phosphate two-phase system are studied in detail. As phase composition increased, demixing time decreased gradually. Phase volume ratio was found to have a significant effect on demixing time at low phase compositions. However, at intermediate and high phase compositions, only a small effect on demixing time was observed. The effect of phase composition and volume ratio on demixing behavior was explained based on the droplet size of the dispersed phase, which is the resultant effect of the physical properties of the phases. At all the phase compositions studied, the acoustically assisted process decreased the demixing time by 17-60% when compared to demixing under gravity alone. Increasing the cell concentration increased the demixing time markedly in case of yeast cells. However, it remained practically constant in the case of Lactobacillus casei cells. Application of an acoustic field reduced the demixing times up to 60% and 40% in the case of yeast and L. casei cells, respectively. Visual observations indicated that ultrasonication caused mild circulation currents in the phase dispersion enhancing droplet-droplet interaction, which in turn enhanced the rate of coalescence, eventually resulting in an enhanced demixing rate.

Acoustic field assisted demixing of aqueous two-phase polymer systems.

Acoustic field assisted demixing was employed to decrease the demixing time in polymer-polymer (polyethylene glycol-maltodextrin) two-phase system. Application of acoustic field has decreased the demixing time in these systems up to 2-fold. Ultrasonication has induced mild circulation currents in the phase dispersion, which has enhanced the rate of droplet coalescence, eventually resulting in decreased demixing time. In polymer-polymer systems, phase demixing was found to depend greatly on which of the phases is continuous and viscosity of the continuous phase was observed to have a strong influence on the movement of the droplets and hence the phase demixing. Addition of NaCl increased the demixing time and presence of E. coli cells did not seem to have any influence on phase demixing.

Acoustic demixing of aqueous two-phase systems.

Aqueous two-phase systems demix slowly due to similar physical properties. This is one of the major drawbacks for their adaptation for industrial scale extraction of enzymes. In the present work, a method to accelerate the demixing rates of these systems, employing a traveling acoustic wave field is reported for the first time. Phase-demixing for three systems, viz. polyethylene glycol (PEG)/sodium sulfate, PEG/potassium phosphate and PEG/maltodextrin were studied. The acoustically assisted process decreased the demixing time significantly (about 2- to 3-fold in PEG/salt systems and about 2-fold in the PEG/maltodextrin system), compared to that in gravity alone. Ultrasonication apparently enhanced the coalescence of the dispersed phase droplets due to the mild circulation currents it caused in the dispersion. This in turn enhanced the rate of demixing due to the increased migration velocity of the larger droplets.