Fast and easy protocol for the purification of recombinant S-layer protein for synthetic biology applications
Online Publication Date2011-06-17
Print Publication Date2011-07
Permanent link to this recordhttp://hdl.handle.net/10754/598314
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AbstractA goal of synthetic biology is to make biological systems easier to engineer. One of the aims is to design, with nanometer-scale precision, biomaterials with well-defined properties. The surface-layer protein SbpA forms 2D arrays naturally on the cell surface of Lysinibacillus sphaericus, but also as the purified protein in solution upon the addition of divalent cations. The high propensity of SbpA to form crystalline arrays, which can be simply controlled by divalent cations, and the possibility to genetically alter the protein, make SbpA an attractive molecule for synthetic biology. To be a useful tool, however, it is important that a simple protocol can be used to produce recombinant wild-type and modified SbpA in large quantities and in a biologically active form. The present study addresses this requirement by introducing a mild and non-denaturing purification protocol to produce milligram quantities of recombinant, active SbpA.
CitationNorville JE, Kelly DF, Knight TF, Belcher AM, Walz T (2011) Fast and easy protocol for the purification of recombinant S-layer protein for synthetic biology applications. Biotechnology Journal 6: 807–811. Available: http://dx.doi.org/10.1002/biot.201100024.
SponsorsThis work was supported by a KAUST Scholar Graduate Research Fellowship (J.E.N.), the SynBERC NSF ERC (www.synberc.org) (J.E.N. and T. F. K.), National Institutes of Health grant GM52580 (S. C. Harrison) and the U.S. Army Research Office through both the Institute for Soldier Nanotechnologies and the Institute for Collaborative Biotechnologies (A. M. B.). T. W. is an investigator in the Howard Hughes Medical Institute.
PubMed Central IDPMC4374430
CollectionsPublications Acknowledging KAUST Support
- Genetic engineering of the S-layer protein SbpA of Lysinibacillus sphaericus CCM 2177 for the generation of functionalized nanoarrays.
- Authors: Badelt-Lichtblau H, Kainz B, Völlenkle C, Egelseer EM, Sleytr UB, Pum D, Ilk N
- Issue date: 2009 May 20
- Lysinibacillus sphaericus S-layer protein toxicity against Culex quinquefasciatus.
- Authors: Lozano LC, Ayala JA, Dussán J
- Issue date: 2011 Oct
- Molecular characterization of the S-layer gene, sbpA, of Bacillus sphaericus CCM 2177 and production of a functional S-layer fusion protein with the ability to recrystallize in a defined orientation while presenting the fused allergen.
- Authors: Ilk N, Völlenkle C, Egelseer EM, Breitwieser A, Sleytr UB, Sára M
- Issue date: 2002 Jul
- A-protein from achromogenic atypical Aeromonas salmonicida: molecular cloning, expression, purification, and characterization.
- Authors: Maurice S, Hädge D, Dekel M, Friedman A, Gertler A, Shoseyov O
- Issue date: 1999 Aug
- Multitechnique study on a recombinantly produced Bacillus halodurans laccase and an S-layer/laccase fusion protein.
- Authors: Ferner-Ortner-Bleckmann J, Schrems A, Ilk N, Egelseer EM, Sleytr UB, Schuster B
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