Fimbrial surface display systems in bacteria: from vaccines to random libraries

Klemm, Per; Schembri, Mark A.

doi:10.1099/00221287-146-12-3025

Cited by 44 publications

(34 citation statements)

References 47 publications

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“…Type 1 fimbriae are thin, thread-like protrusions on the bacterial cell surface and are expressed in 90% of both clinical and environmental K. pneumoniae isolates as well as almost all members of the Enterobacteriaceae (279,280) (Fig. 4).…”

Section: Type 1 and 3 Fimbriaementioning

confidence: 99%

Klebsiella pneumoniae: Going on the Offense with a Strong Defense

Paczosa

Mecsas

2016

Microbiol Mol Biol Rev

1,295

1,345

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SUMMARYKlebsiella pneumoniaecauses a wide range of infections, including pneumonias, urinary tract infections, bacteremias, and liver abscesses. Historically,K. pneumoniaehas caused serious infection primarily in immunocompromised individuals, but the recent emergence and spread of hypervirulent strains have broadened the number of people susceptible to infections to include those who are healthy and immunosufficient. Furthermore,K. pneumoniaestrains have become increasingly resistant to antibiotics, rendering infection by these strains very challenging to treat. The emergence of hypervirulent and antibiotic-resistant strains has driven a number of recent studies. Work has described the worldwide spread of one drug-resistant strain and a host defense axis, interleukin-17 (IL-17), that is important for controlling infection. Four factors, capsule, lipopolysaccharide, fimbriae, and siderophores, have been well studied and are important for virulence in at least one infection model. Several other factors have been less well characterized but are also important in at least one infection model. However, there is a significant amount of heterogeneity inK. pneumoniaestrains, and not every factor plays the same critical role in all virulentKlebsiellastrains. Recent studies have identified additionalK. pneumoniaevirulence factors and led to more insights about factors important for the growth of this pathogen at a variety of tissue sites. Many of these genes encode proteins that function in metabolism and the regulation of transcription. However, much work is left to be done in characterizing these newly discovered factors, understanding how infections differ between healthy and immunocompromised patients, and identifying attractive bacterial or host targets for treating these infections.

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Section: Type 1 and 3 Fimbriaementioning

confidence: 99%

Klebsiella pneumoniae: Going on the Offense with a Strong Defense

Paczosa

Mecsas

2016

Microbiol Mol Biol Rev

1,295

1,345

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“…In mammals, bacterial display of heterologous antigens has already been widely agreed as a more effective mean to achieve the combined vaccines [5][6][7]. However despite its importance, little information has been exploited on the heterologous expression-based combined vaccines against fish diseases.…”

Section: Introductionmentioning

confidence: 98%

Novel Expression System for Combined Vaccine Production in Edwardsiella tarda Ghost and Cadaver Cells

2010

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To develop combined vaccine systems, we have generated Edwardsiella tarda ghosts (ETG) displaying a foreign protein on the outer membrane and also Ed. tarda cadaver (ETC) expressing a heterologous protein in the cytoplasm. Green fluorescent protein (GFP) was used as a model foreign protein. A constitutive promoter (EtPR C28-1) cloned newly from Ed. tarda was used as a promoter for the expression of foreign protein. Comparison of the strength of the new promoter with a commercially available constitutive promoter (P(HCE)) showed higher expression levels of the novel expression system. The N-terminal domain of ice nucleation protein (InaN), an outer membrane protein of Pseudomonas syringae, was used as an anchor motif for surface display of GFP. By transformation of Ed. tarda with the constructed vectors, GFP was successfully expressed on the surface of ETG and in the cytoplasm of ETC. When compared to P(HCE) driven expression, approximately more than 2 times of GFP was expressed on ETG and in ETC by EtPR C28-1 promoter when judged by fluorescent spectrophotometry. Furthermore, significantly higher expression of GFP on the surface of ETG by EtPR C28-1 than by P(HCE) was demonstrated by serum agglutination assay. These results suggest that the newly cloned Ed. tarda constitutive promoter is capable to express foreign proteins not only on the surface of Ed. tarda ghosts but also in the cytoplasm of Ed. tarda cadavers, and can be used as an efficient promoter for the expression of heterologous antigens of the ETG and ETC-based combined vaccines.

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“…The third method of choice for heterologous display on Gram-negative bacteria is the use of highly polymeric surface organelles like fimbriae, or flagella as scaffolds for surface presentation (Klemm and Schembri, 2000b). Fimbriae are adhesive surface organelles which enable bacteria to target and colonize specific host tissues.…”

Section: Surface Display Systems For Gram-negative Bacteriamentioning

confidence: 99%

Engineering of staphylococcal surfaces for biotechnological applications

Wernérus

Lehtiö

Samuelson

et al. 2002

Journal of Biotechnology

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The engineering of bacterial surfaces has in recent years attracted a lot of attention with applications in many different areas of bioscience. Here we describe the use of two different surface display systems for the gram-positive bacteria Staphylococcus carnosus and Staphylococcus xylosus in various biotechnological applications.Environmental microbiology currently attracts a lot of attention since genetically engineered plants and bacteria might be used as bioadsorbents for sequestration of toxic metals. Bacterial surface display of metal-binding peptides might enable recycling of the biomass by desorption of accumulated heavymetals. In an attempt to recruit staphylococcal display systems for bioremediation purposes, polyhistidyl peptides were successfullly displayed on the surface of recombinant S. carnosus and S. xylosus cells. Whole-cell Ni 2+ -binding assays demonstrated that the recombinant cells had gained metal-binding capacity compared to wild-type cells.Tailor-made, metal-binding staphylococci was created using a previously constructed phage-display combinatorial protein library based on a fungal cellulose-binding domain (CBD) derived from the cellobiohydrolase Cel7A of Trichoderma reseii. Novel metal-binding CBDs were generated through a phage mediated selection procedure. Selected CBD variants, now devoid of cellulose binding, were randomly selected and sequence analysis of selected variants revealed a marked preference for histidine residues at the randomized positions. Surface display of these novel CBD variants resulted in recombinant staphylococci with increased metal-binding capacity compared to control strains, indicating that this could become a general strategy to engineer bacteria for improved binding to specific metal ions.Directed immobilization of cells with surface displayed heterologous proteins have widespread use in modern biotechnology. Among other things they could provide a convenient way of generating biofilters, biocatalysts or whole-cell diagnostic devices. It was therefore investigated whether directed immobilization of recombinant staphylococci on cotton fibers could be achieved by functional display of a fungal cellulose-binding domain (CBD). Recombinant S. carnosus cells with surface anchored CBDs from Trichoderma reseii Cel6A were found to efficiently bind to cotton fibers creating almost a monolayer on the fibrous support. The co-expression of this CBD together with previously described metal-binding proteins on the surface of our staphylococci would create means for developing effective bioadsorbents for remediation purposes.The original plasmid vector, designed for heterologous surface display on recombinant S. carnosus cells has exhibited problems related to structural instability, possibly due to the presence of a phage f1 origin of replication in the vector sequence. This would be a problem if using the vector system for library display applications. Therefore, novel surface display vectors, lacking the phage ori were constructed and evaluated by enzymatic a...

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Fimbrial surface display systems in bacteria: from vaccines to random libraries

Cited by 44 publications

References 47 publications

Klebsiella pneumoniae: Going on the Offense with a Strong Defense

Klebsiella pneumoniae: Going on the Offense with a Strong Defense

Novel Expression System for Combined Vaccine Production in Edwardsiella tarda Ghost and Cadaver Cells

Engineering of staphylococcal surfaces for biotechnological applications

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