Interfacial behavior of proteins in three-phase partitioning using salt-containing water/tert-butanol systems

Kiss, Éva; Szamos, J.; Tamás, Boróka; Borbás, Réka

doi:10.1016/s0927-7757(98)00361-6

Cited by 46 publications

(11 citation statements)

References 9 publications

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“…This results in better partitioning of proteins or protein like substances in the middle layer. Due to considerable solubility of ammonium salts in water, partitioning area in the phase diagram (Kiss et al, 1998) is much more in comparison to other salts like sodium, potassium salts. So ammonium salts are used in this study.…”

Section: Effect Of Salts On Sunn Hemp Oil Partitioningmentioning

confidence: 99%

Process optimization for the extraction of oil from Crotalaria juncea using three phase partitioning

Dutta

Sarkar

Mukherjee

2015

Industrial Crops and Products

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Section: Effect Of Salts On Sunn Hemp Oil Partitioningmentioning

confidence: 99%

Process optimization for the extraction of oil from Crotalaria juncea using three phase partitioning

Dutta

Sarkar

Mukherjee

2015

Industrial Crops and Products

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“…In biotechnology, interfaces have been used for the recovery and/or separation of bioparticles and solutes from mixtures. In these applications, aqueous two-phase systems (Andrews et al, 1995;Asenjo et al, 1991;Guereca et al, 1994;HeywoodWaddington et al, 1986;Kula, 1986Kula, , 1993Walker and Lyddiatt, 1999) and other liquid-liquid systems (Borbas et al, 2001;Dennison and Lovrien, 1997;Hoeben et al, 2004;Jauregi et al, 2001Jauregi et al, , 2002Kiss et al, 1998;Pike and Denisson, 1989;Tan and Lovrien, 1972) were used to capture the desired products in the interface between the two liquids. In addition, air flotation which makes use of the adsorption of particles and/or molecules to air bubbles that rise in the liquid phase due to buoyancy, has been applied for the recovery of whole cells (Bahr and Schügerl, 1992;De Dousa et al, 2003;Gahr and Schügerl, 1992;Gaudin, 1975;Wang, 1966, 1967;Kalyuzhnyi et al, 1965;Palmieri et al, 1996;Sadowski and Golab, 1991;Tybussek et al, 1994;Vlaski et al, 1996;Wang et al, 1994).…”

Section: Interfacial Tension Forcementioning

confidence: 99%

Strategy for selection of methods for separation of bioparticles from particle mixtures

Hee

Hoeben

Lans

et al. 2006

Biotech & Bioengineering

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The desired product of bioprocesses is often produced in particulate form, either as an inclusion body (IB) or as a crystal. Particle harvesting is then a crucial and attractive form of product recovery. Because the liquid phase often contains other bioparticles, such as cell debris, whole cells, particulate biocatalysts or particulate by-products, the recovery of product particles is a complex process. In most cases, the particulate product is purified using selective solubilization or extraction. However, if selective particle recovery is possible, the already high purity of the particles makes this downstream process more favorable. This work gives an overview of typical bioparticle mixtures that are encountered in industrial biotechnology and the various driving forces that may be used for particle-particle separation, such as the centrifugal force, the magnetic force, the electric force, and forces related to interfaces. By coupling these driving forces to the resisting forces, the limitations of using these driving forces with respect to particle size are calculated. It shows that centrifugation is not a general solution for particle-particle separation in biotechnology because the particle sizes of product and contaminating particles are often very small, thus, causing their settling velocities to be too low for efficient separation by centrifugation. Examples of such separation problems are the recovery of IBs or virus-like particles (VLPs) from (microbial) cell debris. In these cases, separation processes that use electrical forces or fluid-fluid interfaces show to have a large potential for particle-particle separation. These methods are not yet commonly applied for large-scale particle-particle separation in biotechnology and more research is required on the separation techniques and on particle characterization to facilitate successful application of these methods in industry.

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“…Pigments, lipids and enzyme inhibitors are concentrated in the upper phase, proteins are precipitated in intermediate phase as a salt phase while polar components are enriched in the aqueous phase [27].…”

Section: First Cycle Of Tppmentioning

confidence: 99%

Purification and Recovery of Invertase from Potato Tubers (Solanum tuberosum) by Three Phase Partitioning and Determination of Kinetic Properties of Purified Enzyme

Duman¹

2014

Turk J Bioch

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Objective: Three phase partitioning (TPP) is a simple and economical purification and recovery technique for proteins and also enzymes. In this technique; proteins are separated in the interfacial or lower phase while contaminant parts of crude extract partitioning occur in the upper organic phase. In this study invertase was purified and recovered from potato by TPP. Methods:The highest enzyme recovery of 121% and purification fold 5.67 was seen in the aqueous phase at the 20% ammonium sulfate saturation with 1.0: 1.0 t-butanol ratio at the first cycle of partitioning. The TPP was optimized by adding ammonium sulfate to final concentration at 25% with the constant t-butanol ratio to starting material. Results: Purification fold and activity recovery were found as 12.8 and 156%, respectively. The Km value, optimal temperature and optimal pH of purified enzyme were also determined. Conclusion:The present study showed that, TPP can be an attractive and effective technique for the extraction of invertase from potato. Key Words: TPP, invertase, recovery, purification, potato tuber. Conflict of Interest:The authors declare no conflict of interest. ÖZETAmaç: Üçlü faz ayrılması proteinleri ve enzimlerin geri kazanılması ve saflaştırılması için ekonomik ve basit bir yöntemdir. Bu yöntemde proteinler sulu ya da ara fazda toplanırken; ham enzimin kontamine içeriği üst kısımdaki organik fazda toplanma eğilimindedir. Bu çalışmada patates yumrularında üçlü faz ayrılması ile invertaz saflaştırılması ve geri kazanımı gerçekleş-tirilmiştir. Metod: Üçlü faz ayrımı iki döngüde gerçekleştirilmiştir. birinci döngüde enzim %121 geri kazanım ve 5.67 saflaştırma katsayısı ile %20 amonyum sülfat doygunluğunda ve1 ). The reaction was incubated at 37 ºC for 30 min then was stopped by adding 0.1 ml of DNS (3,5-Dinitrosalicylic acid) reagent. The mixture was heated in a boiling water bath for 5 min. It was then cooled in ice bath and the amount of reducing sugars was measured spectrophotometrically at 540 nm [22]. The data presented for all invertase activity determinations are mean values of duplicate assay. One unit of invertase activity was defined as the amount of enzyme which released 1 µmole of glucose from sucrose per minute at pH 4.7 and 37 ºC. Protein concentrationConcentration of the protein was determined by Bradford method [23] using Coomassie brilliant blue G-250 dye as a reagent and bovine serum albumin (BSA) as standard, by measuring the absorbance at 595nm at 25 ºC. Assays were performed in duplicate and the averages were used in calculations. Three-phase partitioning Effect of t-butanolTPP experiments were carried out by employing various t-butanol ratios (crude extract:t-butanol; 1.0:0.5, 1.0:1.0, 1.0:1.5, 1.0:2.0) with a constant ammonium sulfate saturation at 20% (w/v). The mixture was mixed gently and then allowed to stand for 30 min at 37 ºC. Afterwards the mixture was centrifuged at 5000 rpm for 10 min at +4 ºC to facilitate the separation of phases. The lower aqueous phase and the interfacial phase were coll...

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Interfacial behavior of proteins in three-phase partitioning using salt-containing water/tert-butanol systems

Cited by 46 publications

References 9 publications

Process optimization for the extraction of oil from Crotalaria juncea using three phase partitioning

Process optimization for the extraction of oil from Crotalaria juncea using three phase partitioning

Strategy for selection of methods for separation of bioparticles from particle mixtures

Purification and Recovery of Invertase from Potato Tubers (Solanum tuberosum) by Three Phase Partitioning and Determination of Kinetic Properties of Purified Enzyme

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