Publication Details

Category Text Publication
Reference Category Journals
DOI 10.3389/fbioe.2020.572892
Licence creative commons licence
Title (Primary) Genetic cell-surface modification for optimized foam fractionation
Author Blesken, C.C.; Bator, I.; Eberlein, C. ORCID logo ; Heipieper, H.J. ORCID logo ; Tiso, T.; Blank, L.M.
Source Titel Frontiers in Bioengineering and Biotechnology
Year 2020
Department UBT
Volume 8
Page From art. 572892
Language englisch
Keywords rhamnolipid, 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA), integrated product recovery, foam fractionation, cell surface hydrophobicity, large adhesion protein, flagellum, metabolic engineering
Abstract Rhamnolipids are among the glycolipids that have been investigated intensively in the last decades, mostly produced by the facultative pathogen Pseudomonas aeruginosa using plant oils as carbon source and antifoam agent. Simplification of downstream processing is envisaged using hydrophilic carbon sources, such as glucose, employing recombinant non-pathogenic Pseudomonas putida KT2440 for rhamnolipid or 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA, i.e., rhamnolipid precursors) production. However, during scale-up of the cultivation from shake flask to bioreactor, excessive foam formation hinders the use of standard fermentation protocols. In this study, the foam was guided from the reactor to a foam fractionation column to separate biosurfactants from medium and bacterial cells. Applying this integrated unit operation, the space-time yield (STY) for rhamnolipid synthesis could be increased by a factor of 2.8 (STY = 0.17 gRL/L·h) compared to the production in shake flasks. The accumulation of bacteria at the gas-liquid interface of the foam resulted in removal of whole-cell biocatalyst from the reactor with the strong consequence of reduced rhamnolipid production. To diminish the accumulation of bacteria at the gas-liquid interface, we deleted genes encoding cell-surface structures, focusing on hydrophobic proteins present on P. putida KT2440. Strains lacking, e.g., the flagellum, fimbriae, exopolysaccharides, and specific surface proteins, were tested for cell surface hydrophobicity and foam adsorption. Without flagellum or the large adhesion protein F (LapF), foam enrichment of these modified P. putida KT2440 was reduced by 23 and 51%, respectively. In a bioreactor cultivation of the non-motile strain with integrated rhamnolipid production genes, biomass enrichment in the foam was reduced by 46% compared to the reference strain. The intensification of rhamnolipid production from hydrophilic carbon sources presented here is an example for integrated strain and process engineering. This approach will become routine in the development of whole-cell catalysts for the envisaged bioeconomy. The results are discussed in the context of the importance of interacting strain and process engineering early in the development of bioprocesses.
Persistent UFZ Identifier
Blesken, C.C., Bator, I., Eberlein, C., Heipieper, H.J., Tiso, T., Blank, L.M. (2020):
Genetic cell-surface modification for optimized foam fractionation
Front. Bioeng. Biotechnol. 8 , art. 572892 10.3389/fbioe.2020.572892