Intracellular production of IFN-alpha 2b in Lactococcus lactis

Biotechnol Lett. 2014 Mar;36(3):581-5. doi: 10.1007/s10529-013-1390-4. Epub 2013 Nov 2.

Abstract

Human interferon alpha (IFN-α) was expressed in two strains of Lactococcus lactis by aid of two promoters (P32 and Pnis) giving rise to two recombinant strains: MG:IFN and NZ:IFN, respectively. The expression of IFN was confirmed by ELISA and western blotting. Highest production was achieved using glucose for growth of both recombinant strains with nisin, used for induction of the recombinant strain with Pnis promoter, at 30 ng/ml. The optimum time for MG:IFN was 9 h and for NZ:IFN was 4.5 h. The highest productions by MG:IFN and NZ:IFN were 1.9 and 2.4 μg IFN/l, respectively. Both of the expressed IFNs showed bioactivities of 1.9 × 10(6) IU/mg that were acceptable for further clinical studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Culture Media / chemistry
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression
  • Glucose / metabolism
  • Humans
  • Interferon alpha-2
  • Interferon-alpha / genetics
  • Interferon-alpha / metabolism*
  • Lactococcus lactis / genetics
  • Lactococcus lactis / metabolism*
  • Promoter Regions, Genetic
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Time Factors

Substances

  • Culture Media
  • Interferon alpha-2
  • Interferon-alpha
  • Recombinant Proteins
  • Glucose