Pore engineering as a general strategy to improve protein-based enzyme nanoreactor performance

Area of Science:

• Biotechnology
• Nanotechnology
• Biochemistry

Background:

• Enzyme nanoreactors offer enhanced catalytic activity, stability, and longevity for biotechnological and biomedical uses.
• Existing protein-based nanoreactors face limitations due to poor shell selectivity and reduced enzyme activity.
• A broadly applicable and modular nanoreactor platform is needed.

Approach:

• Engineered a universal enzyme nanoreactor platform using microbial encapsulins, which are self-assembling protein nanocompartments.
• Utilized cryo-electron microscopy for structural characterization of protein shell designs.
• Employed fluorescence polarization assays to evaluate molecular flux and substrate range.

Key Points:

• Developed pore-engineered encapsulins for programmable and selective enzyme packaging.
• Demonstrated improved molecular flux and expanded substrate range in the nanoreactors.
• Highlighted the polymorphic nature of the engineered protein shells.

Conclusions:

• The encapsulin-based nanoreactor platform provides a modular and broadly applicable solution for enzyme encapsulation.
• This platform overcomes limitations of previous protein-based nanoreactors, enhancing enzyme performance.
• Paves the way for advanced biotechnological and biomedical applications of enzyme nanoreactors.