Modification of solubility and heat-induced gelation of Amaranth 11S Globulin by protein engineering
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The primary structure of amaranth 11S globulin (Ah11S) was engineered with the aim to improve its functional properties. Four continuous methionines were inserted in variable region V, obtaining the Ah11Sr 4M construction. Changes on protein structure and surface characteristics were analyzed in silico. Solubility and heat-induced gelation of recombinant amaranth 11S proglobulin (Ah11Sr and Ah11Sr 4M) were compared with the native protein (Ah11Sn) purified from amaranth seed flour. The Ah11Sr 4 M showed the highest surface hydrophobicity, but as consequence the solubility was reduced. At low ionic strength (μ = 0.2) and acidic pH (<4.1), the recombinant proteins Ah11Sr and Ah11Sr 4 M had the highest and lowest solubility values, respectively. All globulins samples formed gels at 90 °C and low ionic strength, but Ah11Sn produced the weakest and Ah11Sr the strongest gels. Differential scanning calorimetry analysis under gel forming conditions revealed only exothermic transitions for all amaranth 11S globulins analyzed. In conclusion, the 3D structure analysis has revealed interesting molecular features that could explain the thermal resistance and gel forming ability of amaranth 11S globulins. The incorporation of four continuous methionines in amaranth increased the hydrophobicity, and self-supporting gels formed had intermediate hardness between Ah11Sn and Ah11Sr. These functional properties could be used in the food industry for the development of new products based on amaranth proteins. © 2013 American Chemical Society.
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The primary structure of amaranth 11S globulin (Ah11S) was engineered with the aim to improve its functional properties. Four continuous methionines were inserted in variable region V, obtaining the Ah11Sr%2b4M construction. Changes on protein structure and surface characteristics were analyzed in silico. Solubility and heat-induced gelation of recombinant amaranth 11S proglobulin (Ah11Sr and Ah11Sr%2b4M) were compared with the native protein (Ah11Sn) purified from amaranth seed flour. The Ah11Sr%2b4 M showed the highest surface hydrophobicity, but as consequence the solubility was reduced. At low ionic strength (μ = 0.2) and acidic pH (<4.1), the recombinant proteins Ah11Sr and Ah11Sr%2b4 M had the highest and lowest solubility values, respectively. All globulins samples formed gels at 90 °C and low ionic strength, but Ah11Sn produced the weakest and Ah11Sr the strongest gels. Differential scanning calorimetry analysis under gel forming conditions revealed only exothermic transitions for all amaranth 11S globulins analyzed. In conclusion, the 3D structure analysis has revealed interesting molecular features that could explain the thermal resistance and gel forming ability of amaranth 11S globulins. The incorporation of four continuous methionines in amaranth increased the hydrophobicity, and self-supporting gels formed had intermediate hardness between Ah11Sn and Ah11Sr. These functional properties could be used in the food industry for the development of new products based on amaranth proteins. © 2013 American Chemical Society.
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Amaranthus hypochondriacus 11S globulins; molecular modeling; recombinant proteins; site directed mutagenesis; thermal-mechanical properties 11S globulin; 3D structure analysis; Exothermic transitions; Functional properties; Site directed mutagenesis; Surface characteristics; Surface hydrophobicity; Thermal mechanical properties; Amino acids; Differential scanning calorimetry; Gelation; Gels; Genetic engineering; Hydrophobicity; Ionic strength; Mechanical properties; Molecular modeling; Solubility; Recombinant proteins; Amaranthus; Amaranthus caudatus; Amaranthus hypochondriacus; gel; globulin; protein intake; recombinant protein; seed storage protein; Amaranthus; chemistry; comparative study; diet supplementation; elemental diet; gel; genetics; heat; metabolism; phase transition; plant seed; protein conformation; protein engineering; protein intake; solubility; surface property; Amaranthus; Dietary Proteins; Dietary Supplements; Food, Formulated; Gels; Globulins; Hot Temperature; Phase Transition; Protein Conformation; Protein Engineering; Recombinant Proteins; Seed Storage Proteins; Seeds; Solubility; Surface Properties
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