2025/10/29 by Zmyslia, Mariia, Capper, Michael J., Grimmeisen, Michael +11
#540 #Article #HeLa cell line #affinity chromatography #amino acid sequence #bioremediation #carboxy terminal sequence #chromatography #controlled study #cryoelectron microscopy #diffusivity #dimer #dimerization #electrophoresis #encapsulation #enzyme activity #human #human cell #hydrodynamics #in vitro study #luminescence #monomer #nanoencapsulation #native polyacrylamide gel electrophoresis #nitroreductase #pH #photon correlation spectroscopy #polyacrylamide gel electrophoresis #prodrug #separation technique #size exclusion chromatography #transmission electron microscopy #virus capsid
paper · doi:10.15488/19885
Nitroreductases are important enzymes for a variety of applications, including cancer therapy and bioremediation. They often require encapsulation to improve stability and activity. We focus on genetically encoded encapsulation of nitroreductases within protein capsids, like encapsulins. Our study showcases the encapsulation of nitroreductase NfsB as functional dimers within encapsulins, which enhances protein activity and stability in diverse conditions. Mutations within the pore region are beneficial for activity of the encapsulated enzyme, potentially by increasing diffusion rates. Cryogenic electron microscopy reveals the overall architecture of the encapsulated dimeric NfsB within the nanoreactor environment and identifies multiple pore states in the shell. These findings highlight the potential of encapsulins as versatile tools for enhancing enzyme performance across various fields.