Expression, purification and activity analysis of human enterokinase light chain from Escherichia coli
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    Abstract:

    [Objective] To express and purify human enterokinase light chain (hEKL) in prokaryotic expression system and detecting its activity in vitro. [Methods] The fragment of hEKL gene was amplified by PCR and cloned into plasmid pMAL-s downstream to the gene of fusion partner MBP-tag. The recombinant plasmid pMAL-s-hEKL was transformed into E. coli BL21(DE3). After induced expression, affinity chromatography was amplified to purify the target protein and Tricine SDS-PAGE was used to analyze the enzymatic activity. [Results] Majority of the fusion protein MBP-hEKL was expressed in soluble form. 40 mg protein was obtained with a purity of more than 97% from 1 L fermentation broth. Activity analysis showed that MBP-hEKL could cleavage the fusion protein with enterokinase recognition site specially and the specific activity was reached to 6.0×105 U/mmol.

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Niu Li-Xi, Li Jiao, Ji Xue-Xue, Li Xia, Li Yan-Jun, Yang Bin-Sheng. Expression, purification and activity analysis of human enterokinase light chain from Escherichia coli[J]. Microbiology China, 2012, 39(11): 1597-1602

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  • Online: November 21,2012
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