Recombinant expression of computationally designed peptide-bundlemers in Escherichia coli

Nairiti J Sinha; Christopher J Kloxin; Jeffery G Saven; Grethe V Jensen; Zvi Kelman; Darrin J Pochan

doi:10.1016/j.jbiotec.2021.03.004

Recombinant expression of computationally designed peptide-bundlemers in Escherichia coli

J Biotechnol. 2021 Mar 20:330:57-60. doi: 10.1016/j.jbiotec.2021.03.004. Epub 2021 Mar 6.

Authors

Nairiti J Sinha¹, Christopher J Kloxin², Jeffery G Saven³, Grethe V Jensen⁴, Zvi Kelman⁵, Darrin J Pochan⁶

Affiliations

¹ Department of Materials Science and Engineering, University of Delaware, Newark, DE, USA; NIST Center for Neutron Research (NCNR), National Institute of Standards & Technology (NIST), Gaithersburg, MD, USA.
² Department of Materials Science and Engineering, University of Delaware, Newark, DE, USA; Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA.
³ Department of Chemistry, University of Pennsylvania, Philadelphia, PA, USA.
⁴ NIST Center for Neutron Research (NCNR), National Institute of Standards & Technology (NIST), Gaithersburg, MD, USA; Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA.
⁵ Biomolecular Labeling Laboratory, National Institute of Standards & Technology (NIST) and Institute for Bioscience and Biotechnology Research (IBBR), Gaithersburg, MD, USA.
⁶ Department of Materials Science and Engineering, University of Delaware, Newark, DE, USA. Electronic address: pochan@udel.edu.

PMID: 33689866
DOI: 10.1016/j.jbiotec.2021.03.004

Abstract

Computational design of fully artificial peptides is extensively researched by material scientists and engineers for the construction of novel nanostructures and biomaterials. Such design has yielded a peptide-based building block or bundlemer, a coiled coil peptide assembly that undergoes further physical-covalent interactions to form 1D, 2D and, potentially, 3D hierarchical assemblies and displays targeted and biomimetic material properties. Recombinant expression is a convenient, flexible tool to synthesize such artificial and modified peptides in large quantities while also enabling economical synthesis of isotopically labeled peptides and longer protein-like artificial peptides. This report describes the protocol for recombinant expression of a 31-amino acid, computationally designed bundlemer-forming peptide in Escherichia coli. Peptide yields of 10 mgs per liter of media were achieved which highlights complementary advantages of recombinant expression technique relative to conventional laboratory-scale synthesis, such as solid-phase peptide synthesis.

Keywords: Bundlemer; Computational design; Escherichia coli; Peptide; Recombinant expression; Self-assembly.

MeSH terms

Biocompatible Materials
Escherichia coli* / genetics
Nanostructures*
Peptides / genetics
Proteins

Substances

Biocompatible Materials
Peptides
Proteins