Autodisplay: efficient bacterial surface display of recombinant proteins
To display a protein or peptide with a distinct function at the surface of a living bacterial cell is a challenging exercise with constantly increasing impact in many areas of biochemistry and biotechnology. Among other systems in Gram-negative bacteria, the Autodisplay system provides striking advantages when used to express a recombinant protein at the surface of Escherichia coli or related bacteria. The Autodisplay system has been developed on the basis of and by exploiting the natural secretion mechanism of the AIDA-I autotransporter protein. It offers the expression of more than 105 recombinant molecules per single cell, permits the multimerization of subunits expressed from monomeric genes at the cell surface, and allows, after transport of an apoprotein to the cell surface, the incorporation of an inorganic prosthetic group without disturbing cell integrity or cell viability. Moreover, whole cells displaying recombinant proteins by Autodisplay can be subjected to high-throughput screening (HTS) methods such as ELISA or FACS, thus enabling the screening of surface display libraries and providing access to directed evolution of the recombinant protein displayed at the cell surface. In this review, the application of the Autodisplay system for the surface display of enzymes, enzyme inhibitors, epitopes, antigens, protein and peptide libraries is summarised and the perspectives of the system are discussed.
KeywordsVaccine Strain Surface Display Secretion Mechanism Live Vaccine Strain Cell Biocatalyst
I thank all individuals who contributed to our work on Autodisplay and, in particular, I thank Ruth Maas for critical reading of the manuscript.
- Benz I, Schmidt MA (1989) Cloning and expression of an adhesin (AIDA-I) involved in diffuse adherence of enteropathogenic Escherichia coli. Infect Immun 57:1506–1511Google Scholar
- Charbit A, Boulain JC, Ryter A, Hofnung M (1986) Probing the topology of a bacterial membrane protein by genetic insertion of a foreign epitope: expression at the cell surface. EMBO J 5:3029–3037Google Scholar
- Hess J, Dreher A, Gentschev I, Goebel W, Ladel C, Miko D, Kaufmann SH (1996) Protein p60 participates in intestinal host invasion by Listeria monocytogenes. Zentralbl Bakteriol 284:263–272Google Scholar
- Klauser T, Pohlner J, Meyer TF (1990) Extracellular transport of cholera toxin B subunit using Neisseria IgA protease beta-domain: conformation-dependent outer membrane translocation. EMBO J 9:1991–1999Google Scholar
- Klauser T, Pohlner J, Meyer TF (1992) Selective extracellular release of cholera toxin B subunit by Escherichia coli: dissection of Neisseria Iga beta-mediated outer membrane transport. EMBO J 11:2327–2335Google Scholar
- Maurer J, Jose J, Meyer TF (1997) Autodisplay: one-component system for efficient surface display and release of soluble recombinant proteins from Escherichia coli. J Bacteriol 179:794–804Google Scholar
- Maurer J, Jose J, Meyer TF (1999) Characterization of the essential transport function of the AIDA-I autotransporter and evidence supporting structural predictions. J Bacteriol 181:7014–7020Google Scholar
- Reetz MT (2000) Evolution in the test tube as a means to create enantioselective enzymes for use in organic synthesis. Sci Prog 83:157–172Google Scholar
- Wilhelm S, Tommassen J, Jaeger KE (1999) A novel lipolytic enzyme located in the outer membrane of Pseudomonas aeruginosa. J Bacteriol 181:6977–6986Google Scholar