title: A Catalytic Nanoreactor Based on in Vivo Encapsulation of Multiple Enzymes in an Engineered Protein Nanocompartment.
abstract: Bacterial protein compartments concentrate and sequester enzymes, thereby regulating biochemical reactions. Here, we generated a new functional nanocompartment in Escherichia coli by engineering the MS2 phage capsid protein to encapsulate multiple cargo proteins. Sequestration of multiple proteins in MS2-based capsids was achieved by SpyTag/SpyCatcher protein fusions that covalently crosslinked with the interior surface of the capsid. Further, the functional two-enzyme indigo biosynthetic pathway could be targeted to the engineered capsids, leading to a 60 % increase in indigo production in vivo. The enzyme-loaded particles could be purified in their active form and showed enhanced long-term stability in vitro (about 95 % activity after seven days) compared with free enzymes (about 5 % activity after seven days). In summary, this engineered in vivo encapsulation system provides a simple and versatile way for generating highly stable multi-enzyme nanoreactors for in vivo and in vitro applications.
authors: Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue WAB 536, Boston, MA, 02115, USA. tobias_giessen@hms.harvard.edu.|Wyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, Boston, MA, 02115, USA. tobias_giessen@hms.harvard.edu.
authors: Tobias W
authors: TW
authors: Giessen
authors: Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue WAB 536, Boston, MA, 02115, USA. pamela_silver@hms.harvard.edu.|Wyss Institute for Biologically Inspired Engineering, Harvard University, 3 Blackfan Circle, Boston, MA, 02115, USA. pamela_silver@hms.harvard.edu.
authors: Pamela A
authors: PA
authors: Silver
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D036022:Capsid Proteins / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D004798:Enzymes / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D017909:Levivirus / Q000737:chemistry* / Q000378:metabolism; D053761:Nanocomposites / Q000737:chemistry*; D015202:Protein Engineering
chemical_list: D001426:Bacterial Proteins; D036022:Capsid Proteins; D004798:Enzymes
keywords: in vivo encapsulation; indigo biosynthesis; nanoreactor; protein engineering; synthetic biology