Applied Microbiology and Biotechnology

, Volume 50, Issue 1, pp 71–76

High-level expression of the angiotensin-converting-enzyme-inhibiting peptide, YG-1, as tandem multimers in Escherichia coli

  • C. J. Park
  • J. H. Lee
  • S.-S. Hong
  • H.-S. Lee
  • S. C. Kim
ORIGINAL PAPER

DOI: 10.1007/s002530051258

Cite this article as:
Park, C., Lee, J., Hong, SS. et al. Appl Microbiol Biotechnol (1998) 50: 71. doi:10.1007/s002530051258

Abstract

To produce a large quantity of the angiotensin-converting-enzyme(ACE)-inhibiting peptide YG-1, which consists of ten amino acids derived from yeast glyceraldehyde-3-phosphate dehydrogenase, a high-level expression was explored with tandem multimers of the YG-1 gene in Escherichia coli. The genes encoding YG-1 were tandemly multimerized to 9-mers, 18-mers and 27-mers, in which each of the repeating units in the tandem multimers was connected to the neighboring genes by a DNA linker encoding Pro-Gly-Arg for the cleavage of multimers by clostripain. The multimers were cloned into the expression vector pET-21b, and expressed in E. coli BL21(DE3) with isopropyl β-d-thiogalactopyranoside induction. The expressed multimeric peptides encoded by the 9-mer, 18-mer and 27-mer accumulated intracellularly as inclusion bodies and comprised about 67%, 25% and 15% of the total proteins in E. coli respectively. The multimeric peptides expressed as inclusion bodies were cleaved with clostripain, and active monomers were purified to homogeneity by reversed-phase high-performance liquid chromatography. In total, 105 mg pure recombinant YG-1 was obtained from 1 l E. coli culture harboring pETYG9, which contained the 9-mer of the YG-1 gene. The recombinant YG-1 was identical to the natural YG-1 in molecular mass, amino acid sequence and ACE-inhibiting activity.

Copyright information

© Springer-Verlag Berlin Heidelberg 1998

Authors and Affiliations

  • C. J. Park
    • 1
  • J. H. Lee
    • 1
  • S.-S. Hong
    • 2
  • H.-S. Lee
    • 2
  • S. C. Kim
    • 1
  1. 1.Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1 Kusong-dong Yusong-gu, Taejon 305-701, Korea Tel.: + 82 42 869 2619 Fax: + 82 42 869 2610 e-mail: sckim@sorak.kaist.ac.krKR
  2. 2.Samyang Genex Research Institute, 63-1 Hwaam-dong, Yusong-gu, Taejon 305-348, KoreaKR