کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1207862 965283 2007 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems
چکیده انگلیسی

The feasibility and generic applicability of directly integrating conventional discrete operations of cell disruption by high pressure homogenizer and the product capture by aqueous two-phase extraction (ATPE) system have been demonstrated for the extraction of intracellular l-asparaginase from E. coli. In a side-by-side comparison with the conventional ATPE process, including cell disruption, centrifugal clarification and following ATPE, purification of l-asparaginase via this novel in situ ATPE process yielded a product of l-asparaginase with a higher specific activity of 94.8 U/(mg protein) and a higher yield of 73.3%, both of which in the conventional ATPE process were 78.6 U/(mg protein) and 52.1%, respectively. In the purification of l-asparaginase (pI = 4.9), product–debris interactions commonly diminish its recovery. It was demonstrated that immediate extraction of l-asparaginase in ATPE systems when it is released at pH 5.0 during cell disruption effectively increased its recovery in the top phase due to the reduced interaction between l-asparaginase and cell debris and the reduced degradation by contaminated protease. In addition, no clarification step and/or disruptate storage are required in this in situ ATPE, which reduced the number of unit operations and thus shortened the overall process time. This novel process has a good potential for the separation of other intracellular biological products.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1147, Issue 1, 13 April 2007, Pages 127–134
نویسندگان
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