Structure modifications of AOT reverse micelles due to protein incorporation

Gochman-Hecht, Hadas; Bianco‐Peled, Havazelet

doi:10.1016/j.jcis.2005.10.027

Cited by 20 publications

(21 citation statements)

References 29 publications

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“…In addition, the W 0 value in reverse micellar system would influence on the aperture size of the solubilized protein molecules. There was a strong relation between the incorporated protein size and the reverse micelle size, when the micelle size was comparable to the protein dimension, the structure of proteins was more affected by the water content [31]. Other factors in reverse micelles, such as pH value and ion strength, etc., would affect the aperture size of 7S and 11S globulins.…”

Section: Sem Images Of 7s and 11s Globulinsmentioning

confidence: 99%

Analysis of the Amino Acids of Soy Globulins by AOT Reverse Micelles and Aqueous Buffer

Zhao

Ling

et al. 2011

Appl Biochem Biotechnol

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The 7S and 11S globulins from soybean proteins using reverse micelle and aqueous buffer extraction methods were characterized by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and scanning electron microscope (SEM), and their amino acid compositions were also evaluated. SDS-PAGE did not show electrophoretic differences between 7S and 11S globulin subunits with two extraction methods. SEM analysis showed that the AOT reverse micelle processing of 7S and 11S globulins induced a reduction of droplet size. Some individual amino acid contents of 7S and 11S globulins using two extraction methods were different, some were similar. In all the samples, the glutamic acid, aspartic acid, and leucine were the dominant amino acids while the cystine and methionine were the first-limiting amino acids. The proportion of essential amino acids to the total amino acids (E/T) of the 7S globulin from aqueous buffer and reverse micelles was similar. While significant differences were obtained in the proportion of E/T of the 11S globulin.

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Section: Sem Images Of 7s and 11s Globulinsmentioning

confidence: 99%

Analysis of the Amino Acids of Soy Globulins by AOT Reverse Micelles and Aqueous Buffer

Zhao

Ling

et al. 2011

Appl Biochem Biotechnol

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“…So, we used a reverse micellar phase composed of 15 mmol/L Triton X-45 and 15 mmol/L Span 80 in n-hexane. The water content kept nearly unchanged at extensive pH values (3)(4)(5)(6)(7)(8)(9)(10)(11) and chelated metal concentration range. 19 Besides the nonionic surfactants, the organic phase contained 0 to 8 mmol/L sodium neutralized HDEHP (NaDEHP), which was prepared as described previously.…”

Section: Nickel-chelate Reverse Micellesmentioning

confidence: 95%

“…[1][2][3][4] Traditionally, ionic surfactants such as bis-2-ethylhexyl sodium sulfosuccinate (AOT) have been mostly used to form reverse micelles. [2][3][4] Solubilization of biomolecules in the ionic surfactant reverse micelles is mainly based on electrostatic interactions between the negatively charge surfactant head group and the positively charge protein molecules. The strong electrostatic interactions often cause two problems associated with the use of ionic reverse micelles for protein separation.…”

Section: Introductionmentioning

confidence: 99%

His‐tagged protein purification by metal‐chelate affinity extraction with nickel‐chelate reverse micelles

Dong

Feng

Sun

2010

Biotechnology Progress

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Di(2-ethylhexyl) phosphoric acid (HDEHP) was used as a transition metal ion chelator and introduced to the nonionic reverse micellar system composed of equimolar Triton X-45 and Span 80 at a total concentration of 30 mmol/L. Ni(II) ions were chelated to the HDEHP dimers in the reverse micelles, forming a complex denoted as Ni(II)R(2). The Ni(II)-chelate reverse micelles were characterized for the purification of recombinant hexahistidine-tagged enhanced green fluorescent protein (EGFP) expressed in Escherichia coli. The affinity binding of EGFP to Ni(II)R(2) was proved by investigation of the forward and back extraction behaviors of purified EGFP. Then, EGFP was purified with the affinity reverse micelles. It was found that the impurities in the feedstock impeded EGFP transfer to the reverse micelles, though they were little solubilized in the organic phase. The high specificity of the chelated Ni(2+) ions toward the histidine tag led to the production of electrophoretically pure EGFP, which was similar to that purified by immobilized metal affinity chromatography. A two-stage purification by the metal-chelate affinity extraction gave rise to 87% recovery of EGFP. Fluorescence spectrum analysis suggests the preservation of native protein structure after the separation process, indicating the system was promising for protein purification.

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“…In the past 3 decades, the most widely studied reverse micelles are formed by ionic surfactants such as bis-2-ethylhexyl sodium sulfosuccinate (AOT). 2,4,5 Solubilization of proteins in the ionic surfactant reverse micelles is mainly based on electrostatic interactions between the negatively charge surfactant head group and the positively charge protein molecules. [7][8][9] Hence, protein solubilization by ionic reverse micelles is highly dependent on the pH and ionic strength of aqueous phase because of the intensive effects of pH and ionic strength on electrostatic interactions.…”

Section: Introductionmentioning

confidence: 99%

A metal‐chelate affinity reverse micellar system for protein extraction

Dong¹,

Meng²,

Feng³

et al. 2009

Biotechnology Progress

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A new nonionic reverse micellar system is developed by blending two nonionic surfactants, Triton X-45 and Span 80. At total surfactant concentrations lower than 60 mmol/L and molar fractions of Triton X-45 less than 0.6, thermodynamically stable reverse micelles of water content (W(0)) up to 30 are formed. Di(2-ethylhexyl) phosphoric acid (HDEHP; 1-2 mmol/L) is introduced into the system for chelating transition metal ions that have binding affinity for histidine-rich proteins. HDEHP exists in a dimeric form in organic solvents and a dimer associated with one transition metal ion, including copper, zinc, and nickel. The copper-chelate reverse micelles (Cu-RM) are characterized for their W(0), hydrodynamic radius (R(h)), and aggregation number (N(ag)). Similar with reverse micelles of bis-2-ethylhexyl sodium sulfosuccinate (AOT), R(h) of the Cu-RM is also linearly related to W(0). However, N(ag) is determined to be 30-90 at W(0) of 5-30, only quarter to half of the AOT reverse micelles. Then, selective metal-chelate extraction of histidine-rich protein (myoglobin) by the Cu-RM is successfully performed with pure and mixed protein systems (myoglobin and lysozyme). The solubilized protein can be recovered by stripping with imidazole or ethylinediaminetetraacetic acid (EDTA) solution. Because various transition metal ions can be chelated to the reverse micelles, it is convinced that the system would be useful for application in protein purification as well as simultaneous isolation and refolding of recombinant histidine-tagged proteins expressed as inclusion bodies.

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Structure modifications of AOT reverse micelles due to protein incorporation

Cited by 20 publications

References 29 publications

Analysis of the Amino Acids of Soy Globulins by AOT Reverse Micelles and Aqueous Buffer

Analysis of the Amino Acids of Soy Globulins by AOT Reverse Micelles and Aqueous Buffer

His‐tagged protein purification by metal‐chelate affinity extraction with nickel‐chelate reverse micelles

A metal‐chelate affinity reverse micellar system for protein extraction

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