Secretory expression of insulin precursor in pichia pastoris and simple procedure for producing recombinant human insulin |
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Authors: | Xie T Liu Q Xie F Liu H Zhang Y |
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Institution: | State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, P. R. China. |
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Abstract: | In this work, Pichia pastoris was applied to produce human insulin by a simple procedure. The synthesized insulin precursor (ILP) gene was inserted into pPIC9K to obtain secretary expression plasmid pPIC9K/ILP. Pichia pastoris GS115 was transformed by pPIC9K/ILP and the high expresser was screened. In a 16 L fermentor, the insulin precursor production was 3.6 g/L. Insulin precursor, purified by one-step chromatography, was converted into human insulin by transpeptidation. The yield of the processing procedure from insulin precursor to insulin reached up to 70%. In vivo assay showed that the biological activity of the produced recombinant human insulin was 28.8 U/mg. |
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