Overexpression for commercial production of recombinant human insulin as A-chain and B-chain fusion protein in Escherichia coli through genetically engineered plasmids.
The construction of expression vectors encoding either the human insulin A- or B-chains fused to a synthetic peptide and the temperature-induced expression of the recombinant genes in Escherichia coli are reported. Using this two-chain approach we also describe the separate isolation of the insulin A- and B-chains from inclusion bodies and their subsequent assembly into native human insulin. The expression yields about 600 mg of the insulin B-chain per litre of culture. Under similar conditions the expression yield of the insulin A-chain corresponds to approximately 500 mg per litre of culture. This is the highest yield from shake flask experiments.
Chakraborty
Chiranjib
C
Department of Biotechnology, Institute of Applied Medicines and Research, Ghaziabad, UP, India. drchiranjib@yahoo.com
D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors; D006801:Humans; D007328:Insulin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
D007328:Insulin; D011993:Recombinant Fusion Proteins