استفاده از پیش‌تیمار مایکروویو در هیدرولیز آنزیمی پروتئین دانه کدو (Cucurbita maxima L.) توسط آلکالاز و بررسی خواص آنتی‌اکسیدانی آن

نوع مقاله : مقاله پژوهشی

نویسندگان

• زینب نوشی منجیلی ¹
• علیرضا صادقی ماهونک ¹
• وحید عرفانی مقدم ²
• محمد قربانی ¹
• هدی شهیری طبرستانی ¹

¹ گروه علوم و صنایع غذایی، دانشکده صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران

² مرکز تحقیقات سرطان، دانشگاه علوم پزشکی گلستان، گرگان، ایران

https://doi.org/10.22067/ifstrj.2023.82861.1264

چکیده

دانه کدو دارای محتوای پروتئین بالایی (40-30 درصد برحسب ماده خشک) است. بخش اعظم دانه­ها به شکل آجیل مورد استفاده قرار می­گیرند و بخشی از آن نیز جزو ضایعات کشاورزی محسوب می­شوند. هیدرولیز شده­های حاصل از پروتئین دانه کدو دارای خواص زیست­فعالی، بویژه فعالیت آنتی­اکسیدانی می­باشند. در این پژوهش محلول محتوی ایزوله پروتئین دانه کدو در معرض پیش­تیمار مایکروویو با توان 900-450 وات به مدت 90-30 ثانیه قرار گرفت و به­عنوان محلول سوبسترا در آزمایشات هیدرولیز آنزیمی استفاده شد. هیدرولیز آنزیمی توسط آلکالاز، با غلظت 5/2-5/0 درصد وزنی نسبت به سوبسترای پروتئینی، در بازه زمانی 20 تا 190 دقیقه، در دما و pH اپتیمم آلکالاز، به­منظور تولید هیدرولیز شده­هایی با پتانسیل آنتی­اکسیدانی انجام گرفت. قدرت آنتی­اکسیدانی با استفاده از روش­های مهار رادیکال آزاد DPPH، قدرت آنتی­اکسیدانی کل و فعالیت شلاته­کنندگی آهن اندازه­گیری شد. نتایج نشان داد، بیشینه فعالیت آنتی­اکسیدانی در شرایط بدون پیش­تیمار، طی زمان 165 دقیقه و نسبت E/S 2/2 درصد با قابلیت مهار رادیکال آزاد DPPH (5/40 درصد)، قدرت آنتی‌اکسیدانی کل (79/0) و شلاته­کنندگی آهن (2/96 درصد) بدست آمد. درحالی‌که با اعمال پیش­تیمار مایکروویو، بیشینه فعالیت آنتی­اکسیدانی در زمانی کوتاه­تر و غلظت آنزیم کمتر (105 دقیقه و نسبت 5/1 درصد E/S)، با قابلیت مهار رادیکال آزاد DPPH (52 درصد)، قدرت آنتی‌اکسیدانی کل (711/0) و شلاته­کنندگی آهن (93 درصد) بدست آمد. بنابراین، می­توان نتیجه گرفت که استفاده از پیش­تیمار مایکروویو در هیدرولیز آنزیمی، علاوه بر دستیابی به هیدرولیز شده­هایی با قابلیت آنتی­اکسیدانی مناسب، موجب صرفه­جویی در زمان و غلظت آنزیم مورد استفاده در طی هیدرولیز نیز می­شود.

کلیدواژه‌ها

• پروتئین هیدرولیز شده
• پیش‌تیمار مایکروویو
• دانه کدو
• فعالیت آنتی‌اکسیدانی

موضوعات

• شیمی مواد غذایی

عنوان مقاله [English]

Using Microwave Pretreatment in the Enzymatic Hydrolysis of Pumpkin Seed Protein (Cucurbita maxima L.) by Alcalase and Investigating Its Antioxidant Properties

نویسندگان [English]

• Zeinab Nooshi Manjili ¹
• Alireza Sadeghi Mahoonak ¹
• Vahid Erfani Moghadam ²
• Mohammad Ghorbani ¹
• Hoda Shahiri Tabarestani ¹

¹ Department of Food Science and Technology, Faculty of Food Industry, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran

² Cancer Research Center, Golestan University of Medical Sciences, Gorgan, Iran

چکیده [English]

Introduction
Seeds and nuts have received increasing attention due to their nutritional value and the high therapeutic properties of their bioactive compounds. Most of the seeds are used as nuts, and some of them are considered agricultural waste. Pumpkin seeds have a high content of protein (30–40% in terms of dry matter). Proteins are among the vital health-giving components that provide nitrogen, essential amino acids and energy necessary for normal cells. Pumpkin seeds are a good source of amino acids such as valine, histidine, isoleucine, leucine, threonine and methionine. Protein hydrolysate is a mixture of peptides and amino acids that can show antioxidant, antimicrobial, anticancer, antidiabetic and antihypertensive properties. During hydrolysis, proteins are broken into small peptides and amino acids. Since enzymatic hydrolysis is performed in relatively mild conditions and no amino acid damage occurs, this type of hydrolysis is preferred over acid and alkaline hydrolysis. Hydrolysates obtained from pumpkin seed protein have bioactive properties, especially antioxidant activity. Pretreatment of proteins before enzymatic hydrolysis acts to improve the release of bioactive peptides from different proteins. Pretreatment can facilitate the unfolding the structure of proteins and thus increase the access of enzymes to peptide bonds. The main properties of microwaves usually show three characteristics: penetration, reflection and absorption. Microwave assisted enzymatic hydrolysis can shorten the time  and improve the speed of the reaction. The purpose of this research was to investigate the antioxidant activity of pumpkin seed protein hydrolysates (Cucurbita maxima L.) by alcalase enzyme in two conditions: without pretreatment and using microwave pretreatment.
 
Material and Methods
In this study, Pumpkin (Cucurbita maxima L.) was purchased from the local market of Astane Ashrafieh in Gilan province. The seeds were scooped manuallyand then dried in an oven at 50°C for 72 hours. After the production of protein concentrate from pumpkin seeds, the chemical properties of the concentrate, such as the amount of fat, protein, ash and moisture, were measured. The isolated pumpkin seed solution was exposed to microwave energy with a power of 450-900 watts for 30–90 seconds and was used as a substrate solution in enzymatic hydrolysis experiments. It should be noted that after measuring the total antioxidantactivityr for different powers and times of microwave pretreatment, the power of 600 watts for 60 seconds was selected  and applied before enzymatic hydrolysis. Enzymatic hydrolysis was done by alcalase enzyme with a concentration of 0.5 to 2.5% compared to the protein substrate during 20 to 190 minutes, and the optimum temperature and pH of alcalase were determined in order to produce hydrolysates with antioxidant activity. Antioxidant activity was measured by using DPPH free radical inhibition, total antioxidant activity and iron chelation activity methods.
 
Result and Discussion
Bioactive peptides produced by the enzymatic hydrolysis of proteins have significant antioxidant properties. Pumpkin seeds can be used as a rich source of nutrients and bioactive compounds in various food industries. The results showed that the maximum amount of antioxidant activity without pre-treatment was achieved in 165 minutes with a 2.2% ratio of E/S by using DPPH free radical scavenging activity (40.5%), total antioxidant power (0.79), and iron chelation activity (96.2%) methods. By using microwave pre-treatment, the maximum amount of antioxidant activity was achieved in a shorter time and with less enzyme (105 minutes and E/S ratio 1.5%) using DPPH free radical scavenging (52%), total antioxidant power (0.711), and iron chelation activity (93%). Therefore, it can be concluded that using microwave assisted enzymatic hydrolysis , in addition to achieving hydrolysates with proper antioxidant activity, is a suitable method to save time and reduce enzyme concentrations used in enzymatic hydrolysis.

کلیدواژه‌ها [English]

• Antioxidant activity
• Microwave pretreatment
• Protein hydrolysates
• Pumpkin seeds

©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.

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