Transport of heterologous proteins to the periplasmic space of Pseudomonas fluorescens using a variety of native signal sequences

Biotechnol Lett. 2007 Oct;29(10):1483-91. doi: 10.1007/s10529-007-9415-5. Epub 2007 May 31.

Abstract

Bacterial expression of recombinant proteins containing disulfide bonds is facilitated by transport of the proteins to the periplasmic space. Several Pseudomonas fluorescens signal sequences have been identified that efficiently direct proteins to the periplasm and provide solubility and yield advantages over the production of proteins fused to the PelB signal sequence in E. coli. For a single chain antibody fragment, the final yield varied from about 1 g/l to 10 g/l when expression in P. fluorescens involved fusion to various P. fluorescens signal sequences.

MeSH terms

  • Blotting, Western
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Genetic Vectors / genetics
  • Periplasm / metabolism*
  • Protein Sorting Signals / genetics
  • Protein Sorting Signals / physiology*
  • Protein Transport
  • Pseudomonas fluorescens / genetics
  • Pseudomonas fluorescens / metabolism*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Sequence Analysis, DNA

Substances

  • Protein Sorting Signals
  • Recombinant Fusion Proteins