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.
Department of Materials Science and Engineering, University of Delaware, Newark, DE, USA; Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA.
J Biotechnol. 2021 Mar 20;330:57-60. doi: 10.1016/j.jbiotec.2021.03.004. Epub 2021 Mar 6.
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.
计算设计的全人工肽是由材料科学家和工程师广泛研究的，用于构建新型纳米结构和生物材料。这种设计产生了基于肽的建筑块或束状聚合物，一种螺旋卷曲的肽组装体，它会进一步发生物理-共价相互作用，形成 1D、2D，并且可能是 3D 分层组装体，并显示出靶向和仿生材料特性。重组表达是一种方便、灵活的工具，可大量合成此类人工和修饰肽，同时还能实现同位素标记肽和更长的类蛋白人工肽的经济合成。本报告描述了在大肠杆菌中重组表达一种 31 个氨基酸的计算设计的束状聚合物形成肽的方案。实现了每升培养基 10 毫克的肽产量，这突出了重组表达技术相对于传统实验室规模合成（如固相肽合成）的互补优势。
