Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system

Biotechnol Lett. 2023 Aug;45(8):1001-1011. doi: 10.1007/s10529-023-03402-x. Epub 2023 Jun 2.

Abstract

Current research focuses on the soluble and high-level expression of biologically active recombinant human IL-29 protein in Escherichia coli. The codon-optimized IL-29 gene was cloned into the Champion™ pET SUMO expression system downstream of the SUMO tag under the influence of the T7 lac promoter. The expression of SUMO-fused IL-29 protein was compared in E. coli Rosetta 2(DE3), Rosetta 2(DE3) pLysS, and Rosetta-gami 2(DE3). The release of the SUMO fusion partner resulted in approximately 98 mg of native rhIL-29 protein with a purity of 99% from 1 l of fermentation culture. Purified rhIL-29 was found to be biologically active, as evaluated by its anti-proliferation assay. It was found that Champion™ pET SUMO expression system can be used to obtained high yield of biologically active soluble recombinant human protein compared to other expression vector.

Keywords: IMAC purification; Interferon-λ1; Interleukin-29; Recombinant protein; Rosetta-gami 2(DE3); pET SUMO.

MeSH terms

  • Codon
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Humans
  • Interleukins* / genetics
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / metabolism

Substances

  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Interleukins
  • Codon