반응표면분석법(RSM)을 이용한 돼지혈장단백질 효소분해조건 최적화

Abstract

As the consumption of meat increases due to the improvement of people's living and the increase of income, the number of slaughtered animals and the slaughter blood from the slaughter process are also increasing. The blood contains more than 28% protein, which contains a lot of essential amino acids such as lysine and non-essential amino acids such as glutamic acid, glycine and alanine. Protein hydrolysates have properties such as inhibitory activity against angiotensin I converting enzyme (ACE), immunologically regulating activity and antioxidant, anti-allergic, antibacterial activities. The purpose of this study was to establish optimal conditions for hydrolysis of porcine plasma proteins using response surface methodology. The response surface methodology was used to optimize the hydrolysis (DH), 2,2-diphenyl-1-picazine (DPPH) radical scavenging activity and Fe2+-chelating activity of the reaction. The results were as follows: 1) The degree of hydrolysis (DH) of the hydrolysate was found to be in the range of 39.10-69.56%. DH showed a strong quadratic (p<0.01) effect on E/S (%) and time. 2) The DPPH radical scavenging activity range of the hydrolysate was determined to be 39.64-64.74%. DPPH radical scavenging activity showed linear (p<0.05) and quadratic (p<0.01) effects on pH. However, the interaction between pH and enzyme concentration relative to substrate, hydrolysis time and enzyme concentration relative to substrate can be neglected. 3) The degree of hydrolysis of Fe2+-chelating activity was ranged from 71.27-84.52%. Fe2+-chelating activity shows strong linea r(p<0.01) and quadratic (p<0.01) effects on E/S (%) and strong quadratic (p<0.01) effects on pH. However, the linear and quadratic effects of hydrolysis time can be neglected (p>0.05). 4) Optimal conditions for hydrolysis of porcine plasma proteins are pH 7.88, E/S (%) 1.06, time 449 min. Under these conditions, DH was 75.32%, DPPH radical scavenging activity was 65.73%, and Fe2+-chelate activity was 82.65%. This condition was confirmed as the optimum enzymatic hydrolysis condition. 5) Hydrolysates treated under optimal conditions showed that proteins with molecular weights greater than 72 kDa were almost degraded. it was confirmed that the degradation to less than 10 kDa protein. 6) The amino acid content of hydrolysate treated under optimal conditions was 27,347 g per 100 g, and the amino acid content contributing to antioxidant activity also increased. In this study, the optimum conditions for the antioxidant activity of protein hydrolysates made with the most commercially available Alcalase and Flavourzyme were established. It was confirmed that the hydrolysate produced under these conditions was mostly degraded to proteins of 10 kDa or less. It was confirmed that the content of amino acids contributing to the antioxidant activity was increased. In addition, the smaller protein that is degraded due to the complex treatment is expected to be well absorbed upon ingestion as well as to increase antioxidant activity, and is expected to be used as an additive and a natural antioxidant in animal and fish feeds. |As the consumption of meat increases due to the improvement of people's living and the increase of income, the number of slaughtered animals and the slaughter blood from the slaughter process are also increasing. The blood contains more than 28% protein, which contains a lot of essential amino acids such as lysine and non-essential amino acids such as glutamic acid, glycine and alanine. Protein hydrolysates have properties such as inhibitory activity against angiotensin I converting enzyme (ACE), immunologically regulating activity and antioxidant, anti-allergic, antibacterial activities. The purpose of this study was to establish optimal conditions for hydrolysis of porcine plasma proteins using response surface methodology. The response surface methodology was used to optimize the hydrolysis (DH), 2,2-diphenyl-1-picazine (DPPH) radical scavenging activity and Fe2+-chelating activity of the reaction. The results were as follows: 1) The degree of hydrolysis (DH) of the hydrolysate was found to be in the range of 39.10-69.56%. DH showed a strong quadratic (p<0.01) effect on E/S (%) and time. 2) The DPPH radical scavenging activity range of the hydrolysate was determined to be 39.64-64.74%. DPPH radical scavenging activity showed linear (p<0.05) and quadratic (p<0.01) effects on pH. However, the interaction between pH and enzyme concentration relative to substrate, hydrolysis time and enzyme concentration relative to substrate can be neglected. 3) The degree of hydrolysis of Fe2+-chelating activity was ranged from 71.27-84.52%. Fe2+-chelating activity shows strong linea r(p<0.01) and quadratic (p<0.01) effects on E/S (%) and strong quadratic (p<0.01) effects on pH. However, the linear and quadratic effects of hydrolysis time can be neglected (p>0.05). 4) Optimal conditions for hydrolysis of porcine plasma proteins are pH 7.88, E/S (%) 1.06, time 449 min. Under these conditions, DH was 75.32%, DPPH radical scavenging activity was 65.73%, and Fe2+-chelate activity was 82.65%. This condition was confirmed as the optimum enzymatic hydrolysis condition. 5) Hydrolysates treated under optimal conditions showed that proteins with molecular weights greater than 72 kDa were almost degraded. it was confirmed that the degradation to less than 10 kDa protein. 6) The amino acid content of hydrolysate treated under optimal conditions was 27,347 g per 100 g, and the amino acid content contributing to antioxidant activity also increased. In this study, the optimum conditions for the antioxidant activity of protein hydrolysates made with the most commercially available Alcalase and Flavourzyme were established. It was confirmed that the hydrolysate produced under these conditions was mostly degraded to proteins of 10 kDa or less. It was confirmed that the content of amino acids contributing to the antioxidant activity was increased. In addition, the smaller protein that is degraded due to the complex treatment is expected to be well absorbed upon ingestion as well as to increase antioxidant activity, and is expected to be used as an additive and a natural antioxidant in animal and fish feeds.