Secretory expression of insulin precursor in pichia pastoris and simple procedure for producing recombinant human insulin.
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.
Xie
T
T
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, P. R. China.
Liu
Q
Q
Xie
F
F
Liu
H
H
Zhang
Y
Y
D006801:Humans; D007328:Insulin / Q000096:biosynthesis* / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
D007328:Insulin; D011994:Recombinant Proteins