Bioinspired enzymatic compartments constructed by spatiotemporally confined in situ self-assembly of catalytic peptide

Abstract

Enzymatic compartments, inspired by cell compartmentalization, which bring enzymes and substrates together in confined environments, are of particular interest in ensuring the enhanced catalytic efficiency and increased lifetime of encapsulated enzymes. Herein, we constructed bioinspired enzymatic compartments (TPE-Q18H@GPs) with semi-permeability by spatiotemporally controllable self-assembly of catalytic peptide TPE-Q18H in hollow porous glucan particles (GPs), allowing substrates and products to pass in/out freely, while enzymatic aggregations were retained. Due to the enrichment of substrates and synergistic effect of catalytic nanofibers formed in the confined environment, the enzymatic compartments exhibited stronger substrate binding affinity and over two-fold enhancement of second-order kinetic constant (kcat/Km) compared to TPE-Q18H nanofibers in disperse system. Moreover, GPs enabled the compartments sufficient stability against perturbation conditions, such as high temperature and degradation. This work opens an intriguing avenue to construct enzymatic compartments using porous biomass materials and has fundamental implications for constructing artificial organelles and even artificial cells.

Unravelling the Mechanism of Elastin-like Polypeptide-Enzyme Fusion Stabilization in Organic Solvents

“…9−11 Commonly used strategies to address enzyme stability and downstream processing challenges rely on compartmentalization via immobilization onto an external support material via entrapment, adsorption, or covalent cross-linking. 6,12,13 A variety of support materials have been used, including synthetic polymers, 14−16 protein cages, 17,18 carbohydrate-based biopolymers, 19,20 and vesicles. 21,22 Unfortunately, these supports often restrict enzyme and substrate mobility, leading to diffusion limitations that can negatively impact reaction rates and increase denaturization rates.…”

“…They can also be comparatively expensive, commonly requiring labile cofactors, and can challenge downstream processing of the reaction product mixtures. Multiple strategies have been employed to overcome these challenges and improve both substrate scope and catalytic performance − ; however, the major bottlenecks remaining for broadened industrial translation are the burden of labor-intensive screening steps, low economic viability, and detailed knowledge of the enzyme mechanism required to guide manipulations of the gene of interest. − Commonly used strategies to address enzyme stability and downstream processing challenges rely on compartmentalization via immobilization onto an external support material via entrapment, adsorption, or covalent cross-linking. ,, A variety of support materials have been used, including synthetic polymers, − protein cages, , carbohydrate-based biopolymers, , and vesicles. , Unfortunately, these supports often restrict enzyme and substrate mobility, leading to diffusion limitations that can negatively impact reaction rates and increase denaturization rates. , When the reaction medium contains high organic solvent content, the most commonly employed method for stabilizing the enzymes in hostile environments such as this is their dispersion within micelle or reverse micellar structures . The most widely used surfactants to stabilize enzymes within micelle or reversed micellar structures of high organic composition are sodium bis­(2-ethylhexyl) sulfosuccinate, Brij 30, or cetyltrimethylammonium bromide. − …”

Uricase Crowding via Polyelectrolyte Layers Coacervation for Carbon Fiber-Based Electrochemical Detection of Uric Acid

“…Enzymatic compartments could be developed through encapsulation in polymer nanoparticles [ 6 ] or surface treatment methodologies, such as immobilization onto Langmuir–Blodgett [ 7 ] or Langmuir–Schaefer films [ 8 ]; the latter, however, require expensive equipment and highly qualified personnel. Alternatively, layer-by-layer (LbL) assembled nanoarchitectures are easily accessible through adsorption of polyelectrolytes on the electrode [ 9 , 10 , 11 , 12 , 13 ], which could avoid a decrease in the mobility of enzyme binding sites [ 14 , 15 , 16 , 17 ].…”