Partitioning of β-galactosidase in aqueous two-phase systems containing polyethyleneglycol and phosphate salts

Khayati, Gholam Reza; Anvari, Masumeh; Shahidi, Nushin

doi:10.1016/j.fluid.2014.11.003

Cited by 25 publications

(12 citation statements)

References 32 publications

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“…Legume lectins such as ConBr and other ConA and Cramoll 1,4 are characterized by having hydrophobic sites and preferentially partitioned to PEG-rich phase. [40,44,45] In the present work, lower MM PEG decreased the hydrophobicity of the top phase and the lectin to migrate to the bottom phase. Similarly, ConA also partitioned to bottom phase using PEG/citrate systems [45] and lectins from the crude extract of C. mollis seeds using PEG/citrate systems.…”

Section: Resultssupporting

confidence: 47%

“…Besides, the partition behavior of protein may be altered with the influence of other proteins and non-proteinaceous substances. [30] ATPS technique has been applied for partition and recovery of protease, [31] antibodies, [32] a-amylase, [25,33] glyco-macropeptide, [34] lipase, [35,36] plasmid DNA, [37] lysozyme, [38] ascorbate oxidase, [39] b-galactosidase, [40] fibrinolytic enzyme, [20,40] and polygalacturonate [41] but also, lectins. [42][43][44][45][46] Statistical design of experiments has been widely used for optimization of extraction/purification of biomolecules in ATPS, [25,47] as it allows the estimating of the significant effects of the extraction variables on the selected responses, as well as their possible interactions.…”

Section: Introductionmentioning

confidence: 99%

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Purification of a lectin from Cratylia mollis crude extract seed by a single step PEG/phosphate aqueous two-phase system

Nascimento

Costa

Soares

et al. 2020

Preparative Biochemistry & Biotechnology

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The partitioning and purification of lectins from the crude extract of Cratylia mollis seeds (Cramoll 1,4) was investigated in aqueous two-phase systems (ATPS). A factorial design model (2 4) was used to evaluate the influence of polyethylene glycol (PEG) molar mass (1500-8000 g/mol), PEG concentration (12.5-17.5% w/w), phosphate (10-15% w/w) concentration, and pH (6-8) on the differential partitioning, purification factor, and yield of the lectin. Polymer and salt concentration were the most important variables affecting partition of lectin and used to find optimum purification factor by experimental Box-Behnken design together with the response surface methodology (RSM). ATPS showed best conditions composed by 13.9% PEG1500, 15.3% phosphate buffer at pH 6, which ensured purification factor of 4.70. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a single band of protein with 26.1 kDa. Furthermore, results demonstrated a thermostable lectin presenting activity until 60 C and lost hemagglutinating activity at 80 C. According to the obtained data it can be inferred that the ATPS optimization using RSM approach can be applied for recovery and purification of lectins.

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Section: Resultssupporting

confidence: 47%

Section: Introductionmentioning

confidence: 99%

Purification of a lectin from Cratylia mollis crude extract seed by a single step PEG/phosphate aqueous two-phase system

Nascimento

Costa

Soares

et al. 2020

Preparative Biochemistry & Biotechnology

View full text Add to dashboard Cite

show abstract

“…In most cases, the system-forming components are inexpensive, non-toxic, non-flammable, and, in some cases, biodegradable and recyclable . They have been applied to the partitioning/extraction/purification of several analytes. − Depending on their structure, charge, pH, and others, s ome combinations of polymers and salts do not result in the ATPS . Theories that explain the formation of the ATPS are still specific to each case, and there is no general theory; therefore, obtaining a new ATPS and the characterization of its phase diagram contribute to the understanding of the driving forces responsible for the phase-separation process.…”

Section: Introductionmentioning

confidence: 99%

Thermodynamics Investigation of Partition Behavior of Uric Acid in Aqueous Two-Phase Systems

et al. 2020

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Uric acid (UA) is an important component in biological matrices, and the development of new methods for extracting/separating UA from several complex matrices is necessary. A viable alternative is the use of an aqueous two-phase system (ATPS), which is an environmentally safe and efficient technique. In this work, an extensive study of the thermodynamic approach of UA partitioning was carried out in an ATPS formed with a polymer, sulfate salts, and water. Initially, the new ATPS formed with polyethylene glycol (400 g mol–1), lithium sulfate, and water was characterized by obtaining the position of the binodal curves and the phase compositions. The components’ segregation increases with the increase in the concentration of the polymer and salt where the top phase (TP) becomes richer in polymer and poorer in electrolyte, and the bottom phase has the inverse behavior. In the range of the pH studied, pH 2.40, 5.40, and 6.60 showed no effect on the binodal curve position and phase compositions, while the temperature (288.15, 298.15, and 308.15 K) evaluation indicated that the phase separation process was entropically driven. Afterward, a study of UA partitioning was carried out in several ATPSs, evaluating the effect of system composition, pH, temperature, and ATPS-forming components on the partition coefficient (K) of the UA. The K values ranged from 1.03 ± 0.04 to 6.05 ± 0.25, indicating a partition preference for the TP for all tie-line length (TLL) values. Furthermore, it is noted that the increase in TLL caused an increase in K, which decreases with increasing the temperature; that is, the partition of uric acid is temperature-dependent, and the phase transfer process of the UA is exothermic. The pH effect study showed that the ionized form of UA has a greater interaction with the components of the TP than that of the molecular form because the K value at pH 6.60 (K = 7.59 ± 0.23) is higher than at pH 2.40 (K = 1.98 ± 0.21), while at pH 5.40 (K = 3.84 ± 0.13), the value is intermediate. This behavior is due to the strong electrostatic interaction between the pseudopolycation, formed by Li+ ions plus polyethylene glycol in TP and the ionized form of UA. Finally, higher K values were obtained for the system formed with polyethylene glycol (400 g mol–1), lithium sulfate, and water. Thus, the balance of interactions between the system components and UA is the driving force that will drive the partition in the ATPS.

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“…The more commonly reported systems are formed by macromolecules and salts, particularly by poly(ethylene oxide) (PEO) − and salts such as sulfates, phosphates, citrates, and tartrates. ,, ATPS containing PEO are mainly used for the partition of water-soluble compounds, since the selective distribution of substances between phases in equilibrium is a direct consequence of interactions that are established between solutes and the constituents that form the phases. As a result, applications of such ATPS are limited and involve the separation of more hydrophilic compounds.…”

Section: Introductionmentioning

confidence: 99%

Phase Diagrams, Densities, and Refractive Indexes of Aqueous Two-Phase Systems Comprising (F68, L64, or PEO1500) + (Ammonium, Sodium, or Potassium Thiocyanate Salts) + Water: Effect of Cation and Type of Macromolecule

Silva¹,

Silva²,

Fontoura³

et al. 2019

J. Chem. Eng. Data

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In extraction procedures, the more commonly used aqueous two-phase systems (ATPS) comprise mainly water, salt, and macromolecule, particularly the macromolecule poly(ethylene oxide) (PEO). However, one limitation of such ATPS is their capacity to separate compounds that are more hydrophobic. One possible solution to overcome this restriction is the use of ATPS formed with triblock copolymers, which are more hydrophobic and therefore enable the extraction of hydrophobic solutes. In addition, the range of applications of ATPS formed with thiocyanate salts can be broader, mainly to extract metal ions. In view of this, equilibrium data were acquired in this work by constructing phase diagrams for ATPS comprising macromolecules [poly(ethylene oxide), PEO, or (poly(ethylene oxide))-(poly(propylene oxide))-(poly(ethylene oxide)) triblock copolymers, F68 or L64] + thiocyanate salts (ammonium, sodium, or potassium) + water at 25.0 °C. The influence of the nature of the cation on the formation of the ATPS was investigated and followed the order K+ > Na+ > NH4 +. The capacity of different macromolecules to enable ATPS formation was also examined and followed the order L64 > F68 > PEO1500. Phase inversion occurred with the (L64 or F68) + NH4SCN + water ATPS, in that the top phase is rich in salt and the bottom phase is rich in macromolecule. This aspect is different in most ATPS that are typically described in the literature.

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Partitioning of β-galactosidase in aqueous two-phase systems containing polyethyleneglycol and phosphate salts

Cited by 25 publications

References 32 publications

Purification of a lectin from Cratylia mollis crude extract seed by a single step PEG/phosphate aqueous two-phase system

Purification of a lectin from Cratylia mollis crude extract seed by a single step PEG/phosphate aqueous two-phase system

Thermodynamics Investigation of Partition Behavior of Uric Acid in Aqueous Two-Phase Systems

Phase Diagrams, Densities, and Refractive Indexes of Aqueous Two-Phase Systems Comprising (F68, L64, or PEO1500) + (Ammonium, Sodium, or Potassium Thiocyanate Salts) + Water: Effect of Cation and Type of Macromolecule

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