In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems

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	In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems
	Jian-Hang Zhu; Xi-Luan Yan; Hong-Jun Chen; Zhi-Hui Wang
	2007
发表期刊	Journal of Chromatography A
卷号	1147 期号:1 页码:127–134
摘要	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.
关键词	Aqueous Two-phase Extraction System In Situ Extraction Cell Disruption Intracellular Protein Process Integration
学科领域	生物基化学品
语种	英语
文献类型	期刊论文
条目标识符	http://ir.qibebt.ac.cn/handle/337004/454
专题	生物基材料组群
推荐引用方式 GB/T 7714	Jian-Hang Zhu,Xi-Luan Yan,Hong-Jun Chen,et al. In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems[J]. Journal of Chromatography A,2007,1147(1):127–134.
APA	Jian-Hang Zhu,Xi-Luan Yan,Hong-Jun Chen,&Zhi-Hui Wang.(2007).In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems.Journal of Chromatography A,1147(1),127–134.
MLA	Jian-Hang Zhu,et al."In situ extraction of intracellular l-asparaginase using thermoseparating aqueous two-phase systems".Journal of Chromatography A 1147.1(2007):127–134.

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