Polyelectrolyte precipitation of β-galactosidase fusions containing poly-aspartic acid tails

Zhao, Jiapeng; Ford, Clark; Glatz, Charles E.; Rougvie, Malcolm A.; Gendel, Steven M.

doi:10.1016/0168-1656(90)90112-o

Cited by 46 publications

(34 citation statements)

References 22 publications

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“…A series of ␤-galactosidases (Zhao et al, 1990) with different lengths of polyasparate fused to the C-termini and T4 lysozymes (DaoPin, 1991) with charge-change point mutations were obtained, and their partition coefficients in aqueous two-phase systems were measured. It was found that the partitioning behavior of T4 lysozyme mutants agreed only qualitatively with Equation (1), while for the ␤-galactosidase fusions, a linear relationship between lnK p and z p was not observed.…”

Section: Introductionmentioning

confidence: 99%

Contribution of protein charge to partitioning in aqueous two-phase systems

Fan¹,

Bakır²,

Glatz³

1998

Biotechnol. Bioeng.

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Section: Introductionmentioning

confidence: 99%

Contribution of protein charge to partitioning in aqueous two-phase systems

Fan¹,

Bakır²,

Glatz³

1998

Biotechnol. Bioeng.

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“…The fusion protein bound strongly to a cation-exchange matrix. Poly aspartic acid tails have also been used for recovery, increasing precipitation of the target protein B-galactosidase by the polyelectrolyte polyethyleneimine (Zhao et al 1990;Parker et al 1990). Similarly, Dalb^ge et al(1987) fused an N-terminal Ala-Glu dipeptide tail to human growth hormone (hGH) and the resultant protein was purified by anion-exchange chromatography.…”

Section: Affinity Tailsmentioning

confidence: 99%

Deletion analysis of the starch-binding domain of Aspergillus glucoamylase

Chen

Coutinho

Nikolov

et al. 1995

Protein Eng Des Sel

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“…1 Second, phase separation of protein-polyelectrolyte complexes offers the possibility of protein purification. [2][3][4] Third, polyelectrolytes can be utilized for immobilization and stabilization of enzymes. 5 Many techniques have been applied to polyelectrolyte-protein complexes, including turbidimetry, viscometry, analytical ultracentrifugation, size-exclusion chromatography, fluorescence spectroscopy, electrophoretic light scattering (ELS), static light scattering, electron-spin resonance, circular dichroism, and dynamic light scattering (QELS).…”

Section: Introductionmentioning

confidence: 99%

Potentiometric Studies of the Interaction of Bovine Serum Albumin and Poly(dimethyldiallylammonium chloride)

Wen

Dubin

1997

Macromolecules

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Potentiometric and turbidimetric titrations were used to study the interaction between bovine serum albumin (BSA) and poly(diallyldimethylammonium chloride) (PDADMAC). Binding between BSA and PDADMAC, which takes place only above some critical initial pH (pHc), leads to a decrease in the pH of solution, indicating that the interactions enhance the dissociation constant Ka of the ionizable groups on BSA and result in an increase of the number of net negative charge on the protein. The pH difference caused by the interaction, ∆pH, decreases with added salt, which indicates that the effect of the interaction on the pKa of BSA increases with a decrease of ionic strength. Protein binding to PDADMAC imposes a stronger influence on the Ka of the carboxylic groups than on the Ka of the imidazolium and ammonium groups. The fraction of BSA bound (fb ) [BSA]b/Cpr) increases with polymer concentration Cp until all BSA are bound. The rate at which fb increases with added polymer at low Cp depends on the initial pH (pHi), consistent with an increase in the binding constant with pHi. Upon a further increase of pH, phase separation occurs at some well-defined point, pHφ, which increases with ionic strength. pHφ depends strongly on Cp at fixed BSA concentration, but only weakly on Cp at constant r ) Cpr/Cp. Phase separation may also be observed upon addition of polymer to BSA at pH > pHφ. In the range of pHc < pH < pHφ, pH titration and turbidimetry reveal the formation of soluble complexes. However, even when phase separation (coacervation) occurs, no corresponding change in the pH titration curve is observed, indicating that the protein's acid-base equilibria are unperturbed by phase separation.

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Polyelectrolyte precipitation of β-galactosidase fusions containing poly-aspartic acid tails

Cited by 46 publications

References 22 publications

Contribution of protein charge to partitioning in aqueous two-phase systems

Contribution of protein charge to partitioning in aqueous two-phase systems

Deletion analysis of the starch-binding domain of Aspergillus glucoamylase

Potentiometric Studies of the Interaction of Bovine Serum Albumin and Poly(dimethyldiallylammonium chloride)

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