Auto-inducing Expression of GST-SUMO-MT Fusion Protein in Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2016.02.030

Abstract

Abstract: The auto-inducing expression of GST-SUMO-MT fusion protein in Escherichia coli BL21(DE3)was studied.In order to increase the protein yield, we optimized both the auto-induction fermentation conditions and medium.Recombinant E.coli was cultured in shake flask, of which the culture condition(time and temperature for induction)and medium composition(tryptone, yeast extract, glycerol, glucose and lactose)were optimized by single factor and orthogonal tests, and bacterial concentration, expression level of soluble target protein were determined.Result showed that the optimal carbon and nitrogen composition ratio in auto induction medium was obtained as followed：2% tryptone, 2% yeast extract, 0.3% glycerol, 0.05% glucose, 0.3% lactose.The optimum fermentation conditions were：cultivating in 37℃ for 3 h firstly, then culturing in 25℃ for 13 h.Under the optimal condition, the expression level of soluble target protein and bacterial concentration were 2.2 and 2.3 times of those in LB medium under induction of IPTG respectively.

Key words: metallothionein, fusion protein, soluble expression, auto-induction

PENG Dong,YANG Wei,WANG Zhao,WANG Xi,LIU Zhong-yu. Auto-inducing Expression of GST-SUMO-MT Fusion Protein in Escherichia coli[J]. Biotechnology Bulletin, 2016, 32(2): 219-224.

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