期刊
CHEMBIOCHEM
卷 17, 期 20, 页码 1931-1935出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201600431
关键词
in vivo encapsulation; indigo biosynthesis; nanoreactor; protein engineering; synthetic biology
资金
- German National Academy of Sciences Leopoldina [LPDS 2014-05]
- Defense Advanced Research Projects Agency (DARPA) [HR0011-14-C-0072]
- Wyss Institute for Biologically Inspired Engineering at Harvard University
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.
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