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Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles
13:16

Characterization of Membrane Transporters by Heterologous Expression in E. coli and Production of Membrane Vesicles

Published on: December 31, 2019

9.7K

Soluble versions of outer membrane cytochromes function as exporters for heterologously produced cargo proteins

Helge M Dietrich1, Miriam Edel2, Thea Bursac2

  • 1Department of Molecular Microbiology and Bioenergetics, Goethe University, Frankfurt, Germany.

Microbial Cell Factories
|December 25, 2019
PubMed

Insights

Outer membrane cytochromes, like MtrF from S. oneidensis, can be truncated and secreted to transport other proteins. This method utilizes the type II secretion pathway for efficient heterologous protein export.

Area of Science:

  • Microbiology
  • Protein Biochemistry
  • Molecular Biology

Background:

  • Outer membrane cytochromes play roles in extracellular electron transfer.
  • Understanding protein secretion pathways is crucial for biotechnology.
  • The type II secretion system is a major pathway for protein export in Gram-negative bacteria.

Purpose of the Study:

  • To investigate the secretion of truncated outer membrane cytochromes.
  • To assess the potential of these truncated proteins as transporters for heterologous proteins.
  • To explore the use of the type II secretion pathway for protein export.

Main Methods:

  • Truncation of the MtrF outer membrane cytochrome.
  • Analysis of protein secretion into culture supernatant.
  • Coupling of cellulase to truncated MtrF to measure export efficiency.
  • Assessment of relative cellulase activity.

Main Results:

  • A truncated MtrF variant, lacking the N-terminal lipid anchor sequence, is efficiently secreted.
  • Further truncation of MtrF by 160 amino acids still allows for its detection in the supernatant.
  • The truncated MtrF successfully facilitated the export of cellulase, as indicated by measurable cellulase activity.

Conclusions:

  • Truncated outer membrane cytochromes can be secreted into the culture medium.
  • These secreted proteins can serve as carriers for heterologously produced proteins.
  • Outer membrane cytochromes can be engineered for protein export via the type II secretion pathway.
Keywords:
CellulaseFusion proteinsOuter membrane cytochromesProtein exportShewanella oneidensisType II secretion system

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