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l-Asparaginase release from Escherichia coli cells with aqueous two-phase micellar systems

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Abstract

A method was proposed to release and separate l-asparaginase (EC 3.5.1.1) from Escherichia coli ATCC 11303 cells with aqueous two-phase micellar systems. The systems were composed of K2HPO4 and Triton X-100. The method combines enzyme release with enzyme purification. The influence of Triton X-100 concentration, K2HPO4 concentration, and pH on the release and partition of l-asparaginase was investigated. Experimental results showed that E. coli cells treated with 9.4% (w/v) K2HPO4 and 15% (w/v) Triton X-100 at 25°C for 15–20 h released nearly 80% of the enzyme. Most of the released enzyme was partitioned to the bottom phase (phosphate-rich phase). The effects of Triton X-100 concentration, K2HPO4 concentration, and pH on cloud point were also studied. Electron micrography indicated that the chemical treatment altered the inner structure of E. coli cells significantly.

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Authors and Affiliations

  1. School of Pharmacy, Shanghai Jiao Tong University, 200030, Shanghai, P. R. China

    Minjun Qin & Fengsheng Zhao

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  1. Minjun Qin
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  2. Fengsheng Zhao
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Correspondence to Fengsheng Zhao.

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Qin, M., Zhao, F. l-Asparaginase release from Escherichia coli cells with aqueous two-phase micellar systems. Appl Biochem Biotechnol 110, 11–21 (2003). https://doi.org/10.1385/ABAB:110:1:11

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  • DOI: https://doi.org/10.1385/ABAB:110:1:11

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