Enzyme–Polyelectrolyte Complexes Boost the Catalytic Performance of Enzymes

Abstract

Understanding interactions between polymers and enzymes to boost enzymatic activity is of high importance for application of enzymes in multicomponent systems, such as laundry, food, pharmaceuticals, or cosmetics. Proteases are widely used in industries and increased performance in the presence of polymers has been reported. Boosting of enzymes activity by polymers and understanding of the molecular principles is of high interest in biomedical and biotechnological applications. A molecular understanding of the boosting effect of poly­(acrylic acid) (PAA) and poly­(l-γ-glutamic acid) (γ-PGA) for a nonspecific subtilisin protease (Protein Database (PDB) ID: 1ST3) was generated through biophysical characterization (fluorescence correlation and circular dichroism spectroscopies, isothermal titration calorimetry), molecular dynamics simulations, and protease reengineering (site-saturation mutagenesis). Our study revealed that enthalpically driven interactions via key amino acid residues close to the protease Ca2+ binding sites cause the boosting effect in protease activity. On the molecular level electrostatic interactions results in the formation of protease-polyelectrolyte complexes. Site-saturation mutagenesis on positions S76, I77, A188, V238, N242, and K245 yielded an increased proteolytic performance against a complex protein mixture (trademark CO-3; up to ∼300% and ∼70%) in the presence of PAA and γ-PGA. Being able to fine-tune interactions between proteins and negatively charged polymers through integrative use of computational design, protein re-engineering and biophysical characterization proved to be an efficient workflow to improve protease performance.

Hyperactivation of L-lactate oxidase by liquid-liquid phase separation

“…First, the interaction of the enzyme with the polymer in the liquid droplets may increase the structural stability of the native enzyme, leading to an increased kcat. [21][22][23] Second, liquid droplets are highly crowded with macromolecules, resulting in the exclusion of water molecules. This crowding stabilizes nonnative structures that differ from those in dilute conditions.…”

Application of different Nanocatalysts in industrial effluent treatment: A review

“…A bottlebrush polymer tightly linked on a solid core of PS was also investigated as an Ag nanoparticle carrier platform. However, the synthesis method is complicated, which can lead to high costs and difficulties in raw materials personnel, and time, restricting broad realistic implementations [113,114].…”

pH/enzyme/light triple‐responsive vesicles from lysine‐based amphiphilic diblock copolymers

“…We suppose prolonging the spacer arm between the p NBC moiety and the main chain can weaken the steric effect and hence make the substrate more accessible to the active site of NTR. Furthermore, it is reported that the enzyme activity can be modulated by the noncovalent interactions between the enzyme and polymer 27–29 . The interactions can be induced by van der Waals forces, electrostatic interactions, hydrophobic interactions, and hydrogen bonding 30 .…”

“…Furthermore, it is reported that the enzyme activity can be modulated by the noncovalent interactions between the enzyme and polymer. [27][28][29] The interactions can be induced by van der Waals forces, electrostatic interactions, hydrophobic interactions, and hydrogen bonding. 30 For example, the complementary electrostatic interactions between a polymer and a protein have been utilized to bring about a binding-induced substrate selectivity to the enzyme.…”