استخراج آنزیم پروتئاز با استفاده از سیستم آبی دو فازی مبتنی بر مایع یونی: مطالعه آزمایشگاهی و مدل سازی ترمودینامیکی

نوع مقاله : مقاله پژوهشی

نویسندگان

گروه مهندسی شیمی، دانشکده فنی و مهندسی، دانشگاه اراک، اراک، ایران

چکیده

هدف: سیستم دو فازی آبی به عنوان یک روش استخراج مایع-مایع به دلیل پتانسیل زیادی برای بازیابی و خالص‌سازی بسیاری از مواد بیولوژیکی مانند پروتئین‌ها، آنزیم‌ها و اسیدهای نوکلئیک مورد توجه قرار گرفته است. مایعات یونی، گروهی از نمک‌های آلی هستند که کاملاً از یون تشکیل شده‌اند و می‌توانند به عنوان یک حلال مناسب برای یک هدف خاص به ویژه فرآیندهای جداسازی پیشنهاد شوند. در بین مایعات یونی مختلف، مشتقات آلکیل سولفات دارای مزایای بسیار خوبی مانند سنتز آسان به روش بدون هالید با ویسکوزیته نسبتا کم و با هزینه مناسب هستند که به عنوان یک مایع یونی مطمئن در سیستم های دو فاز آبی در نظر گرفته می‌شوند. در این مطالعه، جداسازی آنزیم پروتئاز با استفاده از سامانه دو فازی آبی مایع یونی 1-اتیل،3-متیل ایمیدازولیوم اتیل سولفات و نمک سدیم کربنات مورد ارزیابی قرار گرفت. اثر پارامترهایی چون غلظت اولیه آنزیم، غلظت اولیه مایع یونی و غلظت اولیه نمک بر میزان خالص سازی پروتئاز بررسی شد.
مواد و روش: طراحی آزمایش باکس-بنکن به عنوان یکی از روش های سطح-پاسخ بمنظور به حداقل رساندن تعداد آزمایشات و بهینه سازی فرآیند مورد استفاده قرار گرفت.
نتایج: شرایط بهینه برای نسبت آنزیم به مایع یونی برابر 210/0 و نسبت آنزیم به نمک برابر 381/0 بدست آمد که در این شرایط، غلظت آنزیم در فاز بالا (فاز غنی از مایع یونی) و راندمان جداسازی به ترتیب 4 میلی گرم بر لیتر و 94 درصد بدست آمدند.
نتیجه‌گیری: نتایج نشان داد که پروتئین تمایل به استخراج در فاز بالا دارد، و غلظت پروتئین در فاز پایین (فاز غنی از نمک) قابل چشم پوشی بوده است. بعلاوه نتایج آزمایشگاهی تطابق مطلوبی با معادله حالت پیشنهادی NRTL-NRF به عنوان یک معادله با دامنه همبستگی  کوتاه داشتند.

کلیدواژه‌ها

موضوعات


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