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A Method of Targeted Cell Isolation via Glass Surface Functionalization
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Genetic Cell-Surface Modification for Optimized Foam Fractionation.

Christian C Blesken1, Isabel Bator1,2, Christian Eberlein3

  • 1iAMB - Institute of Applied Microbiology, ABBt - Aachen Biology and Biotechnology, RWTH, Aachen University, Aachen, Germany.

Frontiers in Bioengineering and Biotechnology
|November 16, 2020
PubMed
Summary
This summary is machine-generated.

Researchers engineered Pseudomonas putida for enhanced rhamnolipid production by reducing foam-associated bacterial removal. Deleting genes for cell surface structures like flagella and LapF decreased foam enrichment, improving rhamnolipid biosynthesis efficiency.

Keywords:
3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA)cell surface hydrophobicityflagellumfoam fractionationintegrated product recoverylarge adhesion proteinmetabolic engineeringrhamnolipid

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Area of Science:

  • Biotechnology and Bioengineering
  • Microbial Fermentation
  • Metabolic Engineering

Background:

  • Rhamnolipids, biosurfactants produced by Pseudomonas aeruginosa, face downstream processing challenges.
  • Recombinant Pseudomonas putida KT2440 offers a non-pathogenic alternative for rhamnolipid production using hydrophilic carbon sources like glucose.
  • Excessive foam formation during scale-up hinders standard fermentation protocols.

Purpose of the Study:

  • To develop an integrated foam fractionation process for efficient rhamnolipid separation.
  • To engineer Pseudomonas putida KT2440 strains to minimize bacterial accumulation in foam.
  • To enhance rhamnolipid production yield through combined strain and process engineering.

Main Methods:

  • Integrated foam fractionation column for biosurfactant separation from medium and cells.
  • Genetic deletion of genes encoding cell-surface structures (flagellum, fimbriae, LapF, etc.) in P. putida KT2440.
  • Assessment of cell surface hydrophobicity and foam adsorption in modified strains.
  • Bioreactor cultivation of engineered strains for rhamnolipid production.

Main Results:

  • Foam fractionation increased rhamnolipid space-time yield by 2.8-fold (0.17 gRL/L·h) compared to shake flasks.
  • Deletion of flagellum and LapF genes reduced foam enrichment of P. putida KT2440 by 23% and 51%, respectively.
  • Biomass enrichment in foam was reduced by 46% in a non-motile engineered strain.

Conclusions:

  • Integrated strain and process engineering successfully intensified rhamnolipid production from hydrophilic carbon sources.
  • Minimizing bacterial foam enrichment is crucial for efficient whole-cell biocatalyst performance in bioreactors.
  • This approach provides a scalable strategy for developing robust whole-cell catalysts for the bioeconomy.