اثر پیش تیمارهای حرارت و انجماد-یخ زدایی بر هیدرولیز آنزیمی پروتئین عدس به وسیله آنزیم آلکالاز و تولید پپتیدهای آنتی‌اکسیدان

نوع مقاله : مقاله علمی - پژوهشی

نویسندگان

1 دانش آموخته کارشناسی ارشد، گروه علوم و صنایع غذایی، واحد شهر قدس، دانشگاه آزاد اسلامی، تهران، ایران

2 استادیار گروه علوم و صنایع غذایی، واحد شهر قدس، دانشگاه آزاد اسلامی، تهران، ایران

3 استاد پژوهشکده زیست فناوری، سازمان پژوهش های علمی و صنعتی ایران، تهران، ایران

چکیده

مقدمه: فعالیت زیستی محصولات هیدرولیز پروتئینی تحت تاثیر نوع آنزیم، فعالیت آنزیم و شرایط هیدرولیز آنزیمی نظیر دما، زمان و نسبت آنزیم/ سوبسترا و فرآیند پیش تیمار پروتئین می­باشد. فرآیند پیش تیمار می­تواند از طریق تاثیر بر ساختار فضایی پروتئین و افزایش دسترسی آنزیم به باندهای پپتیدی، بر پیشرفت هیدرولیز آنزیمی و تولید پپتیدهای آنتی­اکسیدان موثر باشد.
مواد و روش­ها: پروتئین استخراج شده از عدس ابتدا تحت پیش تیمارهای حرارت (65 ،75، 85 درجه سانتی­گراد به مدت 15 دقیقه) و انجماد-یخ زدایی (3 سیکل انجماد در دمای 20- درجه سانتی­گراد و یخ زدایی در دمای محیط) و هیدرولیز آنزیمی به وسیله آنزیم آلکالاز (با نسبت آنزیم به سوبسترا 90 آنسون / کیلوگرم پروتئین، دمای 55 درجه سانتی­گراد، 3 ساعت) قرار گرفت. در طی زمان، پیشرفت هیدرولیز آنزیمی با روش ارتوفتال آلدئید (OPA) و فعالیت آنتی­اکسیدانی با روش­های مهارکنندگی رادیکال­های DPPH و ABTS مورد بررسی قرار گرفتند و با نمونه کنترل (بدون پیش تیمار) مقایسه شدند.
یافته­ها: پیش تیمار نمونه در دمای 75 درجه سانتی­گراد به مدت 180 دقیقه، بیشترین اثر را در افزایش شدت هیدرولیز آنزیمی نسبت به نمونه­های کنترل بدون پیش تیمار داشت. حداکثر میزان فعالیت مهارکنندگی رادیکال DPPH (57/63 درصد) مربوط به نمونه پیش تیمار شده در دمای 65 درجه سانتی­گراد و حداکثر میزان فعالیت مهار­کنندگی رادیکال ABTS (24/36 درصد) مربوط به نمونه پیش تیمار شده بوسیله فرآیند انجماد-یخ زدایی در زمان 180 دقیقه بود.
نتیجه­ گیری: نتایج نشان دادند که پیش­تیمار حرارتی و انجماد-یخ زدایی قبل از هیدرولیز آنزیمی اثر مثبتی بر پیشرفت هیدرولیز آنزیمی و تولید پپتیدهای آنتی­اکسیدان دارد. بر اساس نتایج حاصل از این تحقیق، فرآیند پیش تیمار حرارتی پروتئین عدس در دمای 65 درجه سانتی­گراد یا انجماد-یخ زدایی و هیدرولیز آنزیمی بوسیله آنزیم آلکالاز روشی موثر در تولید محصول هیدرولیز پروتئین عدس برای کاربرد در فرمولاسیون محصولات غذایی فراسودمند شناخته شد.

کلیدواژه‌ها


عنوان مقاله [English]

The Effect of Heat and Freeze-thaw Pretreatment on the Alcalase Enzymatic Hydrolysis of Lentil Protein and Production of Antioxidant Peptides

نویسندگان [English]

  • P. Ghasemi 1
  • M. Mirzaei 2
  • S. Mirdamadi 3
1 M.Sc. Graduated of the Department of Food Science and Technology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran.
2 Assistant Professor of the Department of Food Science and Technology, Shahr-e-Qods Branch, Islamic Azad University, Tehran, Iran.
3 Professor of the Department of Biotechnology, Iranian Research Organization for Science & Technology (IROST), Tehran, Iran.
چکیده [English]

Introduction: The biological activity of protein hydrolysis products is affected by the enzyme type, protein type and enzymatic hydrolysis conditions including temperature, time and enzyme/substrate ratio and pre-treatment process.The protein pretreatment process can cause improving the enzymatic hydrolysis and production of antioxidant peptides by affecting the spatial structure of the protein and increasing the enzyme access to the peptide bounds.
Materials and Methods: In this study, The protein extracted from lentils was first subjected to heat pretreatment (65,75, 85°C, for 15 min) and freeze-thaw (3 freezing cycles at -20 °C and thawing at room temperature). It was then exposed to hydrolysis for 3 hr by alcalase (with an E/S of 90 AU / kg protein, 55°C). Over time, the progress of enzymatic hydrolysis and antioxidant activity were investigated by O-phthaldialdehyde (OPA) assay and DPPH and ABTS radical scavenging methods and compared with the control sample (without pretreatment).
Results: Pre-treatment at 75°C causes the highest value of free amino groups .The maximum DPPH (63.57%) and ABTS (36.24%) radical scavenging activity were observed respectively, for samples pre-treated at 65°C and by freeze-thaw process.
Conclusion: Heat pretreatment and freezing-thawing before enzymatic hydrolysis have a positive effect on the development of enzymatic hydrolysis and production of antioxidant peptides. Based on the results of this study, the process of heat treatment of lentil protein at 65°C or freezing-thawing and enzymatic hydrolysis by enzyme alcalase was identified as an effective method in the production of lentil protein hydrolysis for use in the formulation of functional foods.

کلیدواژه‌ها [English]

  • Antioxidant peptides
  • Enzymatic Hydrolysis
  • Freeze- Thaw Pretreatment
  • Heat pretreatment
  • Lentil protein
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