Fast and easy protocol for the purification of recombinant S-layer protein for synthetic biology applications

Handle URI:
http://hdl.handle.net/10754/598314
Title:
Fast and easy protocol for the purification of recombinant S-layer protein for synthetic biology applications
Authors:
Norville, Julie E.; Kelly, Deborah F.; Knight, Thomas F.; Belcher, Angela M.; Walz, Thomas
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.
Citation:
Norville 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.
Publisher:
Wiley-Blackwell
Journal:
Biotechnology Journal
Issue Date:
17-Jun-2011
DOI:
10.1002/biot.201100024
PubMed ID:
21681963
PubMed Central ID:
PMC4374430
Type:
Article
ISSN:
1860-6768
Sponsors:
This 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.
Appears in Collections:
Publications Acknowledging KAUST Support

Full metadata record

DC FieldValue Language
dc.contributor.authorNorville, Julie E.en
dc.contributor.authorKelly, Deborah F.en
dc.contributor.authorKnight, Thomas F.en
dc.contributor.authorBelcher, Angela M.en
dc.contributor.authorWalz, Thomasen
dc.date.accessioned2016-02-25T13:18:33Zen
dc.date.available2016-02-25T13:18:33Zen
dc.date.issued2011-06-17en
dc.identifier.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.en
dc.identifier.issn1860-6768en
dc.identifier.pmid21681963en
dc.identifier.doi10.1002/biot.201100024en
dc.identifier.urihttp://hdl.handle.net/10754/598314en
dc.description.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.en
dc.description.sponsorshipThis 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.en
dc.publisherWiley-Blackwellen
dc.subjectElectron microscopyen
dc.subjectNon-denaturing purificationen
dc.subjectS-layer proteinen
dc.subjectSbpAen
dc.subjectSynthetic biologyen
dc.subject.meshSynthetic Biologyen
dc.titleFast and easy protocol for the purification of recombinant S-layer protein for synthetic biology applicationsen
dc.typeArticleen
dc.identifier.journalBiotechnology Journalen
dc.identifier.pmcidPMC4374430en
dc.contributor.institutionSynthetic Biology Working Group, Computer Science and Artificial Intelligence Laboratory, Massachusetts Institute of Technology, Cambridge, MA, USA. norville@csail.mit.eduen
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