Identification, repair and characterization of a benzyl alcohol-inducible promoter for recombinant proteins overexpression in Corynebacterium glutamicum

Enzyme Microb Technol. 2020 Nov:141:109651. doi: 10.1016/j.enzmictec.2020.109651. Epub 2020 Aug 22.

Abstract

Corynebacterium glutamicum is an important industrial organism for the production of a variety of biological commodities. We discovered a promoter encoded by the gene NCgl2319 in C. glutamicum, which could be induced by benzyl alcohol, could be used as an efficient tunable expression system. In initial attempts, this promoter failed to function in a recombinant expression system. This was remedied by extending the original genetic context of the promoter, generating a new version Pcat-B. The Pcat-B transcription initiation site, its critical active regions, and its effect of inducers were fully characterized resulting in tunable expression. This approach proved to be very efficient in producing a pharmaceutical protein, N-terminal pro-brain natriuretic peptide (NT-proBNP). Production of approximately 440.43 mg/L NT-proBNP was achieved with the Pcat-B expression system demonstrating its application for controllable pharmaceutical protein production in C. glutamicum.

Keywords: Benzyl alcohol; Corynebacterium glutamicum; Inducible promoter; Pharmaceutical protein production.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Benzyl Alcohols / metabolism*
  • Corynebacterium glutamicum / genetics*
  • Corynebacterium glutamicum / metabolism
  • Gene Expression Regulation, Bacterial
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Industrial Microbiology
  • Metabolic Engineering
  • Natriuretic Peptide, Brain / genetics
  • Natriuretic Peptide, Brain / metabolism
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Promoter Regions, Genetic*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Bacterial Proteins
  • Benzyl Alcohols
  • Peptide Fragments
  • Recombinant Proteins
  • enhanced green fluorescent protein
  • pro-brain natriuretic peptide (1-76)
  • Natriuretic Peptide, Brain
  • Green Fluorescent Proteins