Engineering Tropism of Pseudomonas putida toward Target Surfaces through Ectopic Display of Recombinant Nanobodies.
Sofía FraileMaría BrionesMónica Revenga-ParraVictor de LorenzoMaría Encarnación LorenzoEsteban Martínez-GarcíaPublished in: ACS synthetic biology (2021)
Gram-negative bacteria are endowed with complex outer membrane (OM) structures that allow them to both interact with other organisms and attach to different physical structures. However, the design of reliable bacterial coatings of solid surfaces is still a considerable challenge. In this work, we report that ectopic expression of a fibrinogen-specific nanobody on the envelope of Pseudomonas putida cells enables controllable formation of a bacterial monolayer strongly bound to an antigen-coated support. To this end, either the wild type or a surface-naked derivative of P. putida was engineered to express a hybrid between the β-barrel of an intimin-type autotransporter inserted in the outer membrane and a nanobody (VHH) moiety that targets fibrinogen as its cognate interaction partner. The functionality of the thereby presented VHH and the strength of the resulting cell attachment to a solid surface covered with the cognate antigen were tested and parametrized with Quartz Crystal Microbalance technology. The results not only demonstrated the value of using bacteria with reduced OM complexity for efficient display of artificial adhesins, but also the potential of this approach to engineer specific bacterial coverings of predetermined target surfaces.
Keyphrases
- biofilm formation
- wild type
- pseudomonas aeruginosa
- staphylococcus aureus
- heat shock
- induced apoptosis
- high resolution
- poor prognosis
- candida albicans
- mental health
- single cell
- physical activity
- escherichia coli
- cell cycle arrest
- endoplasmic reticulum stress
- stem cells
- bone marrow
- mesenchymal stem cells
- human immunodeficiency virus
- long non coding rna
- multidrug resistant
- oxidative stress
- hiv testing
- pi k akt
- antiretroviral therapy
- solid state