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Design of modular polyhydroxyalkanoate scaffolds for protein immobilization by directed ligation.

Biomacromolecules. 2018; 
WongJin Xiang,RehmBernd
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Gene Synthesis The SpyCatcher encoding DNA was synthesized by Genscript (Piscataway, USA) and primers were ordered from Integrated DNA Technologies (San Diego, USA). Get A Quote

摘要

In vivo assembled polyhydroxyalkanoate (PHA) particles have been successfully bioengineered to display foreign protein functions towards high-value applications in medicine and industry. To further expand the design space of PHA particles towards immobilization of various functional proteins, we developed a tunable modular protein immobilization method implementing the SpyCatcher/SpyTag chemistry. We successfully displayed the SpyCatcher protein using translational fusion with the Ralstonia eutropha PHA synthase (PhaC). The SpyCatcher domain displayed on the surface of PHA particles was accessible for cross-linker-free ligation with SpyTag-bearing proteins. We demonstrated tunable protein immobilization of va... More

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