Separation of proteins with polyacrylic acids

Sternberg, M.; Hershberger, D.

doi:10.1016/0005-2795(74)90121-4

Cited by 92 publications

(36 citation statements)

References 11 publications

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“…The culture filtrate (total volume 1.5 1) was cooled to 4°C (this temperature was maintained throughout the purification procedure) and adjusted to pH 4.5 with 5 M acetic acid. Chitosanase was precipitated following a modification of the procedure of Sternberg and Hershberger (1974): a 207o (w/v) solution of polyacrylic acid [average relative molecular mass (Mr) 250,000, Aldrich, Milwaukee, Wis., USA] was added dropwise to a final proportion of 4 mg/mg extracellular proteins. After 30 rain of mixing, the precipitate was collected by centrifugation (11,000 g; 30 min) and resuspended in 300 ml distilled water: 1 M NaOH was added until the pH increased to 8.5.…”

Section: Methodsmentioning

confidence: 99%

“…For unknown reasons, none was satisfactory, resulting in significant loss of enzyme activity. Having established that the N174 chitosanase has a slightly alkaline pI (Fink et al 1991), polyacrylic acid, a precipitant used in large-scale preparations of some industrial enzymes (Sternberg and Hershberger 1974) was successfully tried. Direct addition of polyacrylic acid solution to the culture supernatant (the pH of which varied from 5.8 to 6.5 in different experiments) gave relatively low enzyme recoveries (25-55%).…”

Section: Characterization O F the N174 Strain At The Genus Levelmentioning

confidence: 99%

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Purification and characterization of a chitosanase from Streptomyces N174

Boucher

Dupuy

Vidal

et al. 1992

Appl Microbiol Biotechnol

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A highly efficient chitosanase producer, the actinomycete N174, identified by chemotaxonomic methods as belonging to the genus Streptomyces was isolated from soil. Chitosanase production by N174 was inducible by chitosan or D-glucosamine. In culture filtrates the chitosanase accounted for 50-60% of total extracellular proteins. The chitosanase was purified by polyacrylic acid precipitation, CM-Sepharose and gel permeation chromatography. The maximum velocity of chitosan degradation was obtained at 65 ° C when the pH was maintained at 5.5. The enzyme degraded chitosans with a range of acetylation degrees from 1 to 60% but not chitin or CM-cellulose. The enzyme showed an endo-splitting type of activity and the end-product of chitosan degradation contained a mixture of dimers and trimers of D-glucosamine.

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Section: Methodsmentioning

confidence: 99%

Section: Characterization O F the N174 Strain At The Genus Levelmentioning

confidence: 99%

Purification and characterization of a chitosanase from Streptomyces N174

Boucher

Dupuy

Vidal

et al. 1992

Appl Microbiol Biotechnol

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“…These interactions are modulated by such variables as pH and ionic strength, and may result in soluble complexes, 1,2 complex coacervation, [3][4][5][6] or the formation of amorphous precipitates. [7][8][9] Protein-polyelectrolyte complexation can change the activity of catalytic proteins (enzymes), 10 -12 alter ligand binding to transport proteins, 1,7 and stabilize biological activity against temperature change.…”

Section: Introductionmentioning

confidence: 99%

Light scattering, CD, and ligand binding studies of ferrihemoglobin-polyelectrolyte complexes

et al. 1999

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Quasi-elastic light scattering (QELS), electrophoretic light scattering (ELS), CD spectroscopy, and azide binding titrations were used to study the complexation at pH 6.8 between ferrihemoglobin and three polyelectrolytes that varied in charge density and sign. Both QELS and ELS show that the structure of the soluble complex formed between ferrihemoglobin and poly(diallyldimethylammonium chloride) [PDADMAC] varies with protein concentration. At fixed 1.0 mg/mL polyelectrolyte concentration, protein addition increases complex size and decreases complex mobility in a tightly correlated manner. At 1.0 mg/mL or greater protein concentration, a stable complex is formed between one polyelectrolyte chain and many protein molecules (i.e., an intrapolymer complex) with apparent diameter approximately 2.5 times that of the protein-free polyelectrolyte. Under conditions of excess polyelectrolyte, each of the three ferrihemoglobin-polyelectrolyte solutions exhibits a single diffusion mode in QELS, which indicates that all protein molecules are complexed. CD spectra suggest little or no structural disruption of ferrihemoglobin upon complexation. Azide binding to the ferrihemoglobin-poly(2-acrylamide-2-methylpropanesulfonate)

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“…Some research was conducted to investigate biological phenomena, such as nonspecific long-range effects between proteins and DNA. 1 Another research was done to reach industrial applications, including protein purification, [2][3][4] enzyme immobilization, 5 modulation of a protein affinity for substrates, 6 immunosensing, [7][8][9] and bioactive sensors. 10 Many techniques were applied to study the global structure of protein-polyelectrolyte complexes (PPCs), such as sedimentation, 11,12 turbidimetry, [13][14][15][16] static light scattering, [17][18][19] quasielastic light scattering (QELS), 20,21 and electrophoretic light scattering.…”

Section: Introductionmentioning

confidence: 99%

Potentiometric studies on the interaction between superoxide dismutase and hyaluronic acid

Fan

Wang

Fan

et al. 2009

J of Applied Polymer Sci

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The formation process of soluble complexes and insoluble aggregates between superoxide dismutase (SOD) and hyaluronic acid (HA) was studied using quasielastic light scattering and turbidimetric titration. The electrostatic binding between them was investigated in detail through potentiometric titration and turbidimetric titration carried out from high to low pH. Turbidimetric titration was used to determine the specific pH values at which soluble complex formation was initiated (pH c ) and phase separation occurred (pH / ). An increase of the ionic strength causes a decrease of pH c and pH / . With the increase of HA concentrations, pH / increases but pH c does not vary. The formed ''salt bridges'' between ÀNH þ 3 (SOD) and ACOO À (HA) result in the formation of stable SOD-HA complexes and even aggregates. The necessary condition of electrostatic binding was also given for protein-acidic polyelectrolyte systems.

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Separation of proteins with polyacrylic acids

Cited by 92 publications

References 11 publications

Purification and characterization of a chitosanase from Streptomyces N174

Purification and characterization of a chitosanase from Streptomyces N174

Light scattering, CD, and ligand binding studies of ferrihemoglobin-polyelectrolyte complexes

Potentiometric studies on the interaction between superoxide dismutase and hyaluronic acid

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