High-level production of human growth hormone in Escherichia coli by a simple recombinant process

J Biotechnol. 1998 Jun 30;62(2):143-51. doi: 10.1016/s0168-1656(98)00054-6.

Abstract

Procedures have been devised for producing in Escherichia coli high yields of purified recombinant human growth hormone (hGH), by utilizing N-terminal pentapeptide sequence of human tumor necrosis factor-alpha, histidine tag and enterokinase cleavage site as a fusion partner. The fusion protein was produced as a soluble protein at the beginning of gene expression, but progressively became insoluble in Escherichia coli cytoplasm. The insoluble protein was solubilized by simple alkaline pH shift and purified to near homogeneity by Ni(2+)-chelated affinity chromatography. Following specific enterokinase cleavage, the recombinant hGH was purified by one-step anion exchange chromatography. The ease and speed of this recombinant process, as well as the high productivity, makes it adaptable to the large-scale production of hGH. Moreover, the highly efficient fusion partner could be applied to the production of other therapeutically important proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Circular Dichroism
  • Cloning, Molecular
  • Escherichia coli / metabolism*
  • Human Growth Hormone / biosynthesis*
  • Human Growth Hormone / genetics
  • Humans
  • Molecular Sequence Data
  • Protein Conformation
  • Recombinant Fusion Proteins / biosynthesis
  • Tumor Necrosis Factor-alpha / biosynthesis
  • Tumor Necrosis Factor-alpha / genetics

Substances

  • Recombinant Fusion Proteins
  • Tumor Necrosis Factor-alpha
  • Human Growth Hormone