تاثیر دما بر روی تشکیل کمپلکس آلفا-لاکتالبومین-اولئوروپئین با به‌کارگیری روش‌های طیف‌سنجی و داکینگ مولکولی

نویسندگان
ایمان کاتوزیان 1
سید مهدی جعفری 2
یحیی مقصودلو 3
لیلا کرمی 4
محمد حسن ایکانی 5
1 دانشجوی دکتری، گروه مهندسی مواد و طراحی صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران
2 استاد، گروه مهندسی مواد و طراحی صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران
3 استاد، گروه علوم و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان، ایران
4 استادیار، گروه زیست شناسی سلولی و مولکولی، دانشگاه خوارزمی، دانشکده علوم زیستی، تهران، ایران
5 استاد، گروه صنایع غذایی پژوهشکده فناوری های شیمیایی سازمان پژوهش های علمی و صنعتی ایران ، تهران ، ایران
چکیده
استفاده از حامل‌ها روش مناسبی برای بهبود انتقال و افزایش زیست دسترس‌پذیری ترکیبات زیست فعال می‌باشد. آلفا-لاکتالبومین دومین پروتئین فراوان در آب‌پنیر است که به لحاظ تغذیه‌ای غنی از اسیدهای آمینه ضروری تریپتوفان، لیزین و سیستئین بوده و می­تواند به عنوان حامل مولکول­های غذا و دارو مورد استفاده قرار گیرد. اولئوروپئین ترکیبی زیست فعال با خواص دارویی و ضد اکسایشی می­باشد که به میزان قابل­توجهی در برگ زیتون و به مقدار کمتر در روغن زیتون وجود دارد. هدف از این مطالعه بررسی برهم کنش هم­زمان ترکیب اولئوروپئین با پروتئین آلفا-لاکتالبومین در دماهای مختلف (25، 50 و 72 درجه سانتی­گراد) و pH 7 با استفاده از روش‌های طیف­سنجی فلوئورسانس، فرابنفش و دورنگ نمایی دورانی به همراه شبیه­سازی داکینگ مولکولی بود. مطالعات ناشی از طیف­سنجی فرابنفش و فلوئورسنت افزایش جایگاه و ثابت اتصال را در اثر افزایش دما به سبب تغییر کنفورماسیون و بازآرایی ساختار پروتئین نشان داد. به­علاوه، طیف‌سنجی دورنگ نمایی دورانی افزایش در ساختار صفحه­ای بتا و کاهش در ساختار مارپیچ آلفا را در اثر افزایش دما تا 72 درجه سانتی­گراد نشان داد. در نهایت، شبیه‌سازی داکینگ مولکولی نشان داد که بهترین انرژی پیوندی پیش­بینی شده جهت تشکیل کمپلکس kcal/mol 3/6- بود و ناحیه اتصال نیز بین شیار مارپیچ آلفا و صفحه بتا قرار داشت. نتایج حاصل سبب درک بهتر و عمیق برهمکنش و اتصال پروتئین آلفا-لاکتالبومین و مولکول اولئوروپئین می‌شود که در نهایت در تولید محصولات غذایی فراسودمند مورد استفاده قرار می‌گیرد.

واژه­های کلیدی: آلفا-لاکتالبومین؛ اولئوروپئین، طیف­سنجی، داکینگ مولکولی، محصولات فراسودمند
کلیدواژه‌ها
آلفا-لاکتالبومین
اولئوروپئین
طیف‌سنجی
داکینگ مولکولی
محصولات فراسودمند

عنوان مقاله English

Exploration of the impact of temperature on the formation of α-lactalbumin-oleuropein complex by the application of spectroscopy methods and molecular docking

نویسندگان English

Iman Katouzian 1
Seid Mahdi Jafari 2
Yahya Maghsoudlou 3
Leila Karami 4
Mohammad Hassan Eikani 5
1 Phd Student of food chemistry., Department of Food design materials engineering, Gorgan University of Agricultural Sciences & Natural Resources, Gorgan, Iran.
2 Professor at Department of ., Department of Food design materials engineering, Gorgan University of Agricultural Sciences & Natural ResourcesGorgan, I.R. Iran.
3 Professor at Department of Food Science & Technology, Gorgan University of Agricultural Sciences & Natural ResourcesGorgan, I.R. Iran.
4 Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.
5 Professor of the Department of Chemical Technologies, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran.
چکیده English

The application of carriers is a proper means of improving the transfer and increasing the bioavailability of bioactive compounds. α-lactalbumin is the second major component of whey protein nutritionally consisted of Trp, Lys and Cys residues which can be employed as nutraceutical carriers. Oleuropein is a bioactive compound with pharmaceutical and antioxidant properties which is found abundantly in olive leaves and at lower levels in olive oil. The current study was undertaken to explore the interaction of α-lactalbumin-oleuropein complex at 25, 50 and 72 ˚C at pH 7 by using fluorescent, UV and circular dichroism spectroscopy techniques together with molecular docking. The results from UV and fluorescent studies demonstrate that site and binding constant are increased as the temperature increased due to the change in conformation and rearrangement of protein structure. Moreover, circular dichroism results depicted that α-helix and β-sheet structures are decreased and increased respectively as a result of temperature increase up to 72 ˚C. Ultimately, the molecular docking findings revealed that the best binding energy for complex formation was about -6.3 kCal/mol and the best binding site was between the α-helix and β-sheet cleft. The findings give us useful information regarding the interaction of oleuropein and α-lactalbumin which can be further used to produce functional foods.

Key words: α-lactalbumin; oleuropein; spectroscopy; molecular docking, functional products

کلیدواژه‌ها English

α-lactalbumin
oleuropein
spectroscopy
molecular docking
functional products
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