Immobilizing enzymes by crosslinking biomolecular condensates

Proposed method for immobilizing enzymes by engineering enzyme-RGG fusion proteins, inducing phase separation into biomolecular condensates, and cross-linking condensates.


Invention Summary:

Enzyme immobilization techniques for industrial use often impede the effectiveness of the biocatalyst. A promising alternative method involves utilizing liquid-liquid phase separation (LLPS) of proteins. By appending intrinsically disordered regions to folded proteins, LLPS can be facilitated. When proteins are sequestered in biomolecular condensates, they can be positioned closely enough for effective crosslinking. This approach offers a novel method for enzyme immobilization, as biomolecular condensates are permeable to small molecules, thereby circumventing the mass transfer limitations typically encountered with other immobilization techniques. Moreover, this method employs milder conditions, which help to maximize the residual enzyme activity post-immobilization.

Rutgers researchers have demonstrated using in vitro experiments that biomolecular condensates can be crosslinked into solid microparticles. This method allows for control over microparticle size based on user needs. Additionally, it enables the capture of other proteins into microparticles using specific recruitment motifs and the direct immobilization of enzymes. 

Market Applications:

  • Pharmaceutical industry
  • Cosmetics industry
  • Food and Beverage industry

Advantages:

  • Use of phase separation for sequestering enzymes for crosslinking
  • Enhanced enzyme activity
  • Limited structural distortion of enzymes
  • Alleviation of diffusional limitations

Intellectual Property & Development Status: Provisional application filed. Patent pending. Available for licensing and/or research collaboration. For any business development and other collaborative partnerships, contact:  marketingbd@research.rutgers.edu

Patent Information: