تأثیر پیش‌تیمار فراصوت بر زمان هیدرولیز به وسیله آنزیم پپسین جهت تولید پپتیدهای آنتی‌اکسیدانی از پروتئین قارچ‌خوراکی (Agaricus bisporus)

نوع مقاله : مقاله پژوهشی

نویسندگان

• آیسان ایزانلو
• علیرضا صادقی ماهونک

گروه علوم و صنایع غذایی، دانشگاه علوم کشاورزی ومنابع طبیعی گرگان، گرگان، ایران

https://doi.org/10.22067/ifstrj.2023.78786.1203

چکیده

پپتیدهای زیست‌فعال در واقع بخش‌های پروتئینی خاصی هستند که علاوه ‌بر ارزش غذایی، تأثیرات مثبتی برعملکرد بدن دارند. فراصوت، به­عنوان یک فناوری سبز، و ارزان، به­طور گسترده­ای برای استخراج پروتئین و ترکیبات آنتی­اکسیدانی استفاده می­شود. تیمار با فراصوت باعث تغییر ساختار سه بعدی پروتئین­ها می­شود. در نتیجه، ترکیبی از پیش‌تیمار با فراصوت و هیدرولیز آنزیمی می‌تواند راهی امیدوارکننده برای اصلاح عملکرد پروتئین‌ها باشد. هدف از این پژوهش بررسی اثر زمان هیدرولیز و همچنین تأثیر پیش­تیمار فراصوت بر هیدرولیز آنزیمی پروتئین قارچ دکمه­ای به­وسیله آنزیم پپسین جهت تولید پپتیدهای آنتی­اکسیدان می­باشد. جهت انجام پژوهش ابتدا قارچ به پودر تبدیل و سپس عمل هیدرولیز در زمان­های30-210 دقیقه با نسبت به آنزیم به سوبسترا 1% (وزنی/وزنی نسبت به وزن سوبسترای پروتئینی) و در دمای 40درجه­سانتی­گراد در چهار وضعیت بدون و با پیش­تیمار فراصوت با توان 40، 70 و 100% صورت گرفت. افزایش توان تیمار فراصوت باعث افزایش قابلیت مهار رادیکال­آزاد DPPH، فعالیت شلاته کنندگی یون آهن، ظرفیت آنتی­اکسیدانی­کل و قدرت احیاکنندگی یون آهن در زمان­های هیدرولیز کوتاه­تر گردید. نتایج نشان داد، نمونه­های پیش­تیمار شده با فراصوت با توان 100% نسبت به نمونه­های بدون پیش­تیمار و پیش­تیمار شده با توان40 و 70% بالاترین خاصیت آنتی­اکسیدانی را دارا می­باشند. بنابراین استفاده از پیش­تیمار فراصوت با توان بالا موجب کوتاه نمودن زمان هیدرولیز جهت دستیابی به پپتیدهای با قابلیت آنتی­اکسیدانی بالاتر و افزایش کارآیی هیدرولیز آنزیمی می­گردد. براساس نتایج تیمار فراصوت با توان100% و طی زمان هیدرولیز 60 دقیقه موجب دستیابی محصولی با قابلیت آنتی‌اکسیدانی بالا می­گردد و به‌عنوان تیمارمناسب انتخاب گردید.

کلیدواژه‌ها

• پپتیدهای زیست‌فعال
• پروتئین
• پودر قارچ
• پیش‌تیمار
• ویژگی آنتی‌اکسیدانی

موضوعات

• شیمی مواد غذایی

عنوان مقاله [English]

The Effect of Ultrasound Pretreatment on Hydrolysis Time by Pepsin Enzyme to Produce Antioxidant Peptides from Edible Mushroom (Agaricus bisporus) Protein

نویسندگان [English]

• Iysan Izanloo
• Alireza Sadeghi Mahoonak

Department of Food Science & Technology, Gorgan University of Agricultural Sciences & Natural Resources, Gorgan, Iran

چکیده [English]

Introduction
 Free radicals originate from oxidation reactions decrease food quality and also promote incidence of various diseases such as cancer. In this regard, the use of natural compounds with antioxidant properties, such as bioactive peptides, is of interest to many researchers. Food-derived bioactive peptides, can play an important role in the oxidative systems. Ultrasound, as a cheap and green technology, is widely used to extract proteins and antioxidant compounds. Ultrasound pretreatment before enzymatic hydrolysis can open the protein structure and increase the intensity of proteolysis by increasing the exposure of peptide bonds prone to enzymatic hydrolysis; which increases the production efficiency of bioactive peptides. Ultrasound treatment changes the three-dimensional structure of proteins. Therefore, a combination of pretreatment with ultrasound and sequential enzymatic hydrolysis can be a promising way to modify the function of proteins.
 
Materials and Methods
 In this research the effect of hydrolysis time and ultrasonic pretreatment on enzymatic hydrolysis of edible mushroom protein by pancreatic enzyme to produce peptides with high antioxidant capacity was evaluated. First edible mushroom was turned into powder and then, in order to optimize the production of hydrolyzed proteins with maximum antioxidant activity, the hydrolysis was performed 30, 60, 90, 120, 150, 180 and 210 minutes with a ratio of enzyme to substrate of 1% (based on the result of previous research) and at 40°C in four conditions (1- without ultrasound pre-treatment, 2- with ultrasound pre-treatment with 40% power, 3- with ultrasound pre-treatment with 70% power and 4- with ultrasound pre-treatment with 100% power) by ultrasound probe in 5 minutes before adding the enzyme. In the next step, the antioxidant capacity of hydrolyzed proteins was measured at different times by DPPH free radical scavenging activity, iron ion reduction power, iron ion chelation and total antioxidant capacity.
 
Results
 The results showed that the highest DPPH free radical scavenging activity in untreated and treated samples with 40, 70 and 100% ultrasound power were 69.1, 77.45, 79.07 and 80.27, respectively. In most of the hydrolysis times, DPPH free radical scavenging activity in ultrasound treatment with 100% power was higher than the samples treated with 40 and 70% power. The highest total antioxidant capacity in untreated and treated samples with 40, 70 and 100% ultrasound power were 0.871, 1.025, 1.05 and 1.2 (absorption at 695 nm), respectively. In most of the hydrolysis times, the total antioxidant capacity in the samples treated with ultrasound with 100% power was higher than the samples treated with 40 and 70% power. The results showed that the highest reducing power of Fe³⁺ in untreated and treated samples with 40, 70 and 100% ultrasound power were 2.03, 2.40, 2.44 and 2.51(absorption at 700 nm), respectively. The highest iron ion chelation power in untreated and treated samples with 40, 70 and 100% ultrasound power were 25.22, 30.40, 26.52 and 41.10%, respectively. By increasing the ultrasound power in most of the hydrolysis times, the chelating power of iron ions in the ultrasound treatment with 100% power was higher than the samples pretreated with 40 and 70% power. The results showed that samples pretreated with 100% power ultrasound have the highest antioxidant properties compared to samples without pretreatment and pretreated with 40% and 70% ultrasound power. Based on the results, using ultrasound treatment with 100% power and during hydrolysis time of 60 minutes, a product with high antioxidant capacity was obtained and selected as a suitable treatment.
 
Conclusion
 The ultrasonic mechanism is attributed to its thermal effects, cavitation and mechanical efficiency, so that it can increase the mass transfer and increase the contact between the substrate and the enzyme or change the spatial structure of the substrate. The results showed that samples pretreated with ultrasound with 100% power have the highest antioxidant properties compared to samples without pretreatment and pretreated with 40 and 70% power. Therefore, the use of high-power ultrasonic pretreatment shortens the hydrolysis time to achieve peptides with higher antioxidant capacity and thus increases the efficiency of enzymatic hydrolysis.

کلیدواژه‌ها [English]

• Antioxidant properties
• Bioactive peptides
• Mushroom powder
• Pretreatment
• Protein

©2023 The author(s). This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source.

مراجع

1. Abadía-García, L., Castaño-Tostado, E., Ozimek, L., Romero-Gómez, S., Ozuna, C., & Amaya-Llano, S.L. (2016). Impact of ultrasound pretreatment on whey protein hydrolysis by vegetable proteases. Innovative Food Science & Emerging Technologies, 37, 84-90. https://doi.org/10.1016/j.ifset.2016.08.010
2. Aderinola, T.A., Fagbemi, T.N., Enujiugha, V.N., Alashi, A.M., & Aluko, R.E. (2019). In vitro antihypertensive and antioxidative properties of alcalase‐derived Moringa oleifera seed globulin hydrolysate and its membrane fractions. Journal of Food Processing and Preservation, 43(2), e13862. https://doi.org/10.1111/jfpp.13862
3. Bhat, Z.F., Kumar, S., & Bhat, H.F. (2015). Bioactive peptides of animal origin: a review. Journal of Food Science and Technology, 52(9), 5377-5392. https://doi.org/10.1007/s13197-015-1731-5
4. Bougatef, A., Hajji, M., Balti, R., Lassoued, I., Triki-Ellouz, Y., & Nasri, M. (2009). Antioxidant and free radical-scavenging activities of smooth hound (Mustelus mustelus) muscle protein hydrolysates obtained by gastrointestinal proteases. Food Chemistry, 114(4), 1198-1205. https://doi.org/10.1016/j.foodchem.2008.10.075
5. Chen, L., Chen, J., Ren, J., & Zhao, M. (2011). Effects of ultrasound pretreatment on the enzymatic hydrolysis of soy protein isolates and on the emulsifying properties of hydrolysates. Journal of Agricultural and Food Chemistry, 59(6), 2600-2609. https://doi.org/10.1021/jf103771x
6. Chi, C.F., Hu, F.Y., Wang, B., Li, T., & Ding, G.F. (2015). Antioxidant and anticancer peptides from the protein hydrolysate of blood clam (Tegillarca granosa) muscle. Journal of Functional Foods, 15, 301-313. https://doi.org/10.1016/j.jff.2015.03.045
7. Dasgupta, N., & De, B. (2007). Antioxidant activity of some leafy vegetables of India: A comparative study. Food Chemistry, 101(2), 471-474. https://doi.org/10.1016/j.foodchem.2006.02.003
8. Ding, Q., Zhang, T., Niu, S., Cao, F., Wu-Chen, R.A., Luo, L., & Ma, H. (2018). Impact of ultrasound pretreatment on hydrolysate and digestion products of grape seed protein. Ultrasonics Sonochemistry, 42, 704-713. https://doi.org/10.1016/j.ultsonch.2017.11.027
9. FitzGerald, R.J., & Meisel, H. (2000). Milk protein-derived peptide inhibitors of angiotensin-I-converting enzyme. British Journal of Nutrition, 84(S1), 33-37. https://doi.org/10.1017/S0007114500002221
10. Guerra-Almonacid, C.M., Torruco-Uco, J.G., Murillo-Arango, W., Méndez-Arteaga, J.J., & Rodríguez-Miranda, J. (2019). Effect of ultrasound pretreatment on the antioxidant capacity and antihypertensive activity of bioactive peptides obtained from the protein hydrolysates of Erythrina edulis. Emirates Journal of Food and Agriculture, 288-296. https://doi.org/10.1016/j.ultsonch.2019.104787
11. He, J. Z., Ru, Q.M., Dong, D.D., & Sun, P.L. (2012). Chemical characteristics and antioxidant properties of crude water soluble polysaccharides from four common edible mushrooms. Molecules, 17(4), 4373-4387. https://doi.org/10.3390/molecules17044373
12. Horwitz, W., Chichilo, P., & Reynolds, H. (1970). Official methods of analysis of the Association of Official Analytical Chemists. Official Methods of Analysis of the Association of Official Analytical Chemists.
13. Jamdar, S.N., Rajalakshmi, V., Pednekar, M.D., Juan, F., Yardi, V., & Sharma, A.(2010). Influence of degree of hydrolysis on functional properties, antioxidant activity and ACE inhibitory activity of peanut protein hydrolysate. Food Chemistry, 121(1), 178-184. https://doi.org/10.1016/j.foodchem.2009.12.027
14. Je, J.Y., Lee, K.H., Lee, M.H., & Ahn, C.B. (2009). Antioxidant and antihypertensive protein hydrolysates produced from tuna liver by enzymatic hydrolysis. Food Research International, 42(9), 1266-1272. https://doi.org/10.1016/j.foodres.2009.06.013
15. Kadam, S.U., Tiwari, B.K., Álvarez, C., & O'Donnell, C.P. (2015). Ultrasound applications for the extraction, identification and delivery of food proteins and bioactive peptides. Trends in Food Science & Technology, 46(1), 60-67. https://doi.org/10.1016/j.tifs.2015.07.012
16. Klompong, V., Benjakul, S., Kantachote, D., & Shahidi, F. (2007). Antioxidative activity and functional properties of protein hydrolysate of yellow stripe trevally (Selaroides leptolepis) as influenced by the degree of hydrolysis and enzyme type. Food Chemistry, 102(4), 1317-1327. https://doi.org/10.1016/j.foodchem.2006.07.016
17. Lavi, I., Nimri, L., Levinson, D., Peri, I., Hadar, Y., & Schwartz, B. (2012). Glucans from the edible mushroom Pleurotus pulmonarius inhibit colitis-associated colon carcinogenesis in mice. Journal of Gastroenterology, 47(5), 504-518. https://doi.org/10.1007/s00535-011-0514-7
18. Li, X.R., Chi, C.F., Li, L., & Wang, B. (2017). Purification and identification of antioxidant peptides from protein hydrolysate of scalloped hammerhead (Sphyrna lewini) cartilage. Marine Drugs, 15(3), 61. https://doi.org/10.3390/md15030061
19. Liang, Q., Ren, X., Ma, H., Li, S., Xu, K., & Oladejo, A.O. (2017). Effect of low-frequency ultrasonic-assisted enzymolysis on the physicochemical and antioxidant properties of corn protein hydrolysates. Journal of Food Quality, 2017. https://doi.org/10.1155/2017/2784146
20. Matmaroh, K., Benjakul, S., Prodpran, T., Encarnacion, A.B., & Kishimura, H. (2011). Characteristics of acid soluble collagen and pepsin soluble collagen from scale of spotted golden goatfish (Parupeneus heptacanthus). Food Chemistry, 129(3), 1179-1186. https://doi.org/10.1016/j.foodchem.2011.05.099
21. Meshginfar, N., Sadeghi, M.A., Ziaiifar, A.M., Ghorbani, M., & Kashaninejad, M. (2014). Optimization of the production of protein hydrolysates from meat industry by products by response surface methodology. Journal of Food Research, 24(2), 215-225.
22. Nourmohammadi, E., Sadeghi Mahoonak, A., Ghorbani, M., Alami, M., & Sadeghi, M. (2017). The optimization of the production of anti-oxidative peptides from enzymatic hydrolysis of Pumpkin seed protein. Iranian Food Science and Technology Research Journal, 13(1), 14-26. https://doi.org/22067/ifstrj.v1395i0.45423
23. Nadeem, M., Ubaid, N., Qureshi, T.M., Munir, M., & Mehmood, A. (2018). Effect of ultrasound and chemical treatment on total phenol, flavonoids and antioxidant properties on carrot-grape juice blend during storage. Ultrasonics Sonochemistry, 45, 1-6. https://doi.org/10.1016/j.ultsonch.2018.02.034
24. Paisansak, S., Sangtanoo, P., Srimongkol, P., Saisavoey, T., Reamtong, O., Choowongkomon, K., & Karnchanata, A. (2020). Angiotensin-I converting enzyme inhibitory peptide derived from the shiitake mushroom (Lentinula edodes). Journal Food Science and Technology, 58(1), 85–97. https://doi.org/ 10.1007/s13197-020-04517-z
25. Pan, X., Zhao, Y.Q., Hu, F.Y., & Wang, B. (2016). Preparation and identification of antioxidant peptides from protein hydrolysate of skate (Raja porosa) cartilage. Journal of Functional Foods, 25, 220-230. https://doi.org/10.1016/j.jff.2016.06.008
26. Sun, Q., Shen, H., & Luo, Y. (2011). Antioxidant activity of hydrolysates and peptide fractions derived from porcine hemoglobin. Journal of Food Science and Technology, 21, 6646-6652. https://doi.org/10.1007/s13197-010-0115-0
27. Vioque, J., Clemente, A., Pedroche, J., Yust, M.D.M., & Millán, F. (2001). Obtención y aplicaciones de hidrolizados proteicos. Grasas y Aceites, 52(2), 132-136. https://doi.org/3989/gya.2001.v52.i2.385
28. Wali, A., Ma, H., Shahnawaz, M., Hayat, K., Xiaong, J., & Jing, L. (2017). Impact of power ultrasound on antihypertensive activity, functional properties, and thermal stability of rapeseed protein hydrolysates. Journal of Chemistry, 2017. https://doi.org/10.1155/2017/4373859
29. Walters, M.E. (2019). Effects of Ultrasonication on the Antioxidant and Anti-diabetic Properties of Hydrolyzed Oat Proteins(Doctoral dissertation, Carleton University). https://doi.org/10.22215/etd/2019-13844
30. Wen, C., Zhang, J., Zhang, H., Dzah, C. S., Zandile, M., Duan, Y., et al. (2018a). Advances in ultrasound assisted extraction of bioactive compounds from cash crops-A review. Ultrasonics Sonochemistry, 48, 538–549. https://doi.org/10.1016/j.ultsonch.2018.07.018
31. Yang, X., Li, Y., Li, S., Oladejo, A. O., Wang, Y., Huang, S., ... & Ye, X. (2017). Effects of low power density multi-frequency ultrasound pretreatment on the enzymolysis and the structure characterization of defatted wheat germ protein. Ultrasonics Sonochemistry, 38, 410-420. https://doi.org/10.1016/j.ultsonch.2017.03.001
32. Yu, L., Sun, J., Liu, S., Bi, J., Zhang, C., & Yang, Q. (2012). Ultrasonic-assisted enzymolysis to improve the antioxidant activities of peanut (Arachin conarachin) antioxidant hydrolysate. International Journal of Molecular Sciences, 13(7), 9051-9068. https://doi.org/10.3390/ijms13079051
33. Zhang, Y., Ma, L., Cai, L., Liu, Y., & Li, J. (2017). Effect of combined ultrasonic and alkali pretreatment on enzymatic preparation of angiotensin converting enzyme (ACE) inhibitory peptides from native collagenous materials. Ultrasonics Sonochemistry, 36, 88-94. https://doi.org/10.1016/j.ultsonch.2016.11.008
34. Zhou, C., Hu, J., Yu, X., Yagoub, A.E.A., Zhang, Y., Ma, H., ... & Otu, P.N.Y. (2017). Heat and/or ultrasound pretreatments motivated enzymolysis of corn gluten meal: Hydrolysis kinetics and protein structure. LWT, 77, 488-496. https://doi.org/10.1016/j.lwt.2016.06.048
35. Zhu, K.X., Su, C.Y., Guo, X.N., Peng, W., & Zhou, H.M. (2011). Influence of ultrasound during wheat gluten hydrolysis on the antioxidant activities of the resulting hydrolysate. International Journal of Food Science & Technology, 46(5), 1053-1059. https://doi.org/10.1111/j.1365-2621.2011.02585.x
36. Zhu, L., Chen, J., Tang, X., & Xiong, Y.L. (2008). Reducing, radical scavenging, and chelation properties of in vitro digests of alcalase-treated zein hydrolysate. Journal of Agricultural and Food Chemistry, 56(8), 2714-2721. https://doi.org/10.1021/jf703697e
37. Zou, Y., Yang, H., Li, P.P., Zhang, M. H., Zhang, X.X., Xu, W.M., & Wang, D.Y. (2019). Effect of different time of ultrasound treatment on physicochemical, thermal, and antioxidant properties of chicken plasma protein. Poultry Science, 98(4), 1925-1933. https://doi.org/10.3382/ps/pey502