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Isolation of pure aminoacids from wheat bran
Sloupová, Klára ; Diviš, Pavel (referee) ; Pořízka, Jaromír (advisor)
Wheat bran is a promising material containing a wide range of useful components, including proteins. In addition, it is produced in significant volumes. Currently, wheat bran is used for the production of energy by combustion and for feed purposes. Gradually, new methods of valorization of this material are being sought. One of the possibilities of using wheat bran is the isolation of proteins, hydrolysis, and separation of selected amino acids. This diploma thesis deals with this issue, it is focused on the recovery of arginine and leucine from a protein isolate. Proteins were extracted from wheat bran by changing the pH. Thanks to the subsequent lyophilization a protein isolate was gained. Prior to hydrolysis of the resulting isolate, a stability test of arginine and leucine amino acid standards was first performed, to which various hydrolysis methods were applied. Acid hydrolysis using a mineralizer, which was applied to the protein isolate, was proved to be the most effective. This was followed by the derivatization of the hydrolysates with OPA and analysis of the resulting hydrolysates by high-performance liquid chromatography with UV-VIS detection. Then, suitable adsorption and desorption conditions were optimized. It was found that the time dependence does not affect the amount of adsorbed material on the sorbent. Therefore, an application time of 15 minutes was chosen. While optimizing the amount of used standard, it was found that the optimal weight was 0.25 g of sorbent. The selected conditions were applied to the protein hydrolyzate. Two fractions were obtained by the separation of selected amino acids due to the change in the pH of the citrate buffer. After the application of this procedure, 0.26 g of arginine and 0.82 g of leucine were obtained from one kilogram after evaporation. From evaporation two, 1.01 g of arginine and 0.25 g of leucine were obtained after evaporation.
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Isolation of pure aminoacids from wheat bran
Sloupová, Klára ; Diviš, Pavel (referee) ; Pořízka, Jaromír (advisor)
Wheat bran is a promising material containing a wide range of useful components, including proteins. In addition, it is produced in significant volumes. Currently, wheat bran is used for the production of energy by combustion and for feed purposes. Gradually, new methods of valorization of this material are being sought. One of the possibilities of using wheat bran is the isolation of proteins, hydrolysis, and separation of selected amino acids. This diploma thesis deals with this issue, it is focused on the recovery of arginine and leucine from a protein isolate. Proteins were extracted from wheat bran by changing the pH. Thanks to the subsequent lyophilization a protein isolate was gained. Prior to hydrolysis of the resulting isolate, a stability test of arginine and leucine amino acid standards was first performed, to which various hydrolysis methods were applied. Acid hydrolysis using a mineralizer, which was applied to the protein isolate, was proved to be the most effective. This was followed by the derivatization of the hydrolysates with OPA and analysis of the resulting hydrolysates by high-performance liquid chromatography with UV-VIS detection. Then, suitable adsorption and desorption conditions were optimized. It was found that the time dependence does not affect the amount of adsorbed material on the sorbent. Therefore, an application time of 15 minutes was chosen. While optimizing the amount of used standard, it was found that the optimal weight was 0.25 g of sorbent. The selected conditions were applied to the protein hydrolyzate. Two fractions were obtained by the separation of selected amino acids due to the change in the pH of the citrate buffer. After the application of this procedure, 0.26 g of arginine and 0.82 g of leucine were obtained from one kilogram after evaporation. From evaporation two, 1.01 g of arginine and 0.25 g of leucine were obtained after evaporation.
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