Antioxidant evaluation of hydrolyzed pumpkin seed protein under the influence of simple and sequential enzymatic hydrolysis using pepsin and trypsin

Document Type : Complete scientific research article

Authors

1 MSc graduate, Department of Food Science and Technology, Islamic Azad University of Azadshahr Branch, Azadshahr, Iran.

2 Assistant Professor, Food Science and Technology Research Center of east Golestan, Islamic Azad University of Azadshahr Branch, Azadshahr, Iran

Abstract

Abstract
Background and objectives: The use of synthetic antioxidants in the food industry is subject to strict laws due to the potential negative effects of these compounds on the human health. As a result, replacing synthetic chemical antioxidants with various natural antioxidant compounds has become an important field of study. Numerous researches have shown that the addition of hydrolyzed proteins or peptides with antioxidant properties can effectively prevent the oxidation of fats during transportation and storage, which preserves the taste and quality of nutrition. Due to the fact that different enzymes have different effects on the same substrate, and due to the fact that it has been claimed that the combination of enzymes improves the bioactivity characteristics of the product, the aim of this research was to compare the effect of simple enzymatic hydrolysis versus sequential hydrolysis on the antioxidant properties of hydrolyzed pumpkin seed protein.
Materials and methods: Pumpkin seed protein was first extracted and then hydrolysed by two enzymes, trypsin and pepsin, in a simple and sequential hydrolysis condition. At first, in order to achieve the appropriate time of simple hydrolysis, the two mentioned enzymes were used separately in the condition of enzyme to substrate ratio of 2%, optimum temperature (37°C) and optimum pH for each enzyme and during the times of 30, 60, 90, 120, 150, 180 and 210 minutes. In the next step, using the optimal time of 180 minute, four sequential hydrolysis treatments were performed.
Results: The results of first step showed that the hydrolyzates produced in 180 minutes have a high antioxidant capacity based on DPPH free radical scavenging activity, iron ion reduction power, total antioxidant capacity and iron ion chelating activity. In the next step, using optimaltime 180 minutes, fpour sequential hydrolysis condition were used as follows: 90 minutes of hydrolysis with trypsin and then 90 minutes with pepsin enzyme (treatment 1), 90 minutes of hydrolysis with pepsin and then 90 minutes with trypsin enzyme (treatment 2), 120 minutes of hydrolysis with trypsin and then 60 minutes with pepsin enzyme (treatment 3), 120 minutes of hydrolysis with pepsin and then 60 minutes with trypsin enzyme (treatment 4) and were compared with two simple hydrolysis treatments including: 180 minutes of hydrolysis with Trypsin (treatment 5) and 180 minutes of treatment with pepsin (treatment 6) in terms of the four above mentioned antioxidant tests. The results showed that the sample under sequential hydrolysis treatments showed higher antioxidant capacity compared to simple hydrolysis.
Conclusion: Due to the difference in the composition and sequence and different size as a result of the different degree of hydrolysis of the peptides, sample resulting from the sequential hydrolysis of the two enzymes showed higher antioxidant activity.

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References

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Food Processing and Preservation Journal
Volume 16, Issue 4
January 2025
Pages 1-19

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