title: Fast and easy protocol for the purification of recombinant S-layer protein for synthetic biology applications.
abstract: A 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.
authors.affiliation: Synthetic Biology Working Group, Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, MA, USA. norville@csail.mit.edu
authors.forename: Julie E
authors.initials: JE
authors.lastname: Norville
authors.forename: Deborah F
authors.initials: DF
authors.lastname: Kelly
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Knight
authors.forename: Angela M
authors.initials: AM
authors.lastname: Belcher
authors.forename: Thomas
authors.initials: T
authors.lastname: Walz
mesh_terms: D001406:Bacillaceae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D003001:Cloning, Molecular; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D016624:Negative Staining; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D058615:Synthetic Biology
chemical_list: D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C055764:S-layer proteins