Expression of soluble cloned porcine pepsinogen A in Escherichia coli

Biochem J. 1996 Apr 15;315 ( Pt 2)(Pt 2):443-6. doi: 10.1042/bj3150443.

Abstract

A system for the production of soluble porcine pepsinogen A (EC 3.4.23.1) was developed by fusing the pepsinogen and thioredoxin genes and then expressing the fused product (Trx-PG) in Escherichia coli. The expressed fusion protein was purified using a combination of ion-exchange and hydrophobic chromatography. Trypsin digestion of the fusion protein yielded pepsinogen which was one residue longer than the intrinsic length. Acidification of either the fusion protein or pepsinogen (tryptic digestion of Trx-PG) yielded recombinant pepsin A (r-pepsin). When compared with commercial porcine pepsin A, r-pepsin had similar milk-clotting and proteolytic activities, kinetic parameters and pH dependency. The above results indicate that an expression system was developed which yielded fully active soluble pepsin(ogen) from Escherichia coli.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular
  • Escherichia coli / genetics*
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Sequence Data
  • Oligopeptides / chemistry
  • Pepsin A / genetics
  • Pepsin A / isolation & purification
  • Pepsinogens / biosynthesis*
  • Pepsinogens / chemistry
  • Pepsinogens / genetics*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Solubility
  • Substrate Specificity
  • Swine
  • Thioredoxins / biosynthesis
  • Thioredoxins / chemistry
  • Thioredoxins / genetics
  • Trypsin

Substances

  • Oligopeptides
  • Pepsinogens
  • Recombinant Fusion Proteins
  • Thioredoxins
  • Trypsin
  • Pepsin A