In Situ Immobilization of Multi-Enzymes for Enhanced Substrate Channeling of Enzyme Cascade Reactions: A Nanoarchitectonics Approach by Directed Metal–Organic Frameworks

Abstract

Rationally tailoring a controlled spatial organization of enzymes in a nanoarchitecture for multienzyme cascade reactions can enhance the catalytic efficiency via substrate channeling. However, attaining substrate channeling is a grand challenge, requiring sophisticated techniques. Herein, we report facile polymerdirected metal−organic framework (MOF)-based nanoarchitechtonics for realizing a desirable enzyme architecture with significantly enhanced substrate channeling. The new method involves the use of poly(acrylamide-co-diallyldimethylammonium chloride) (PADD) as a modulator in a one-step process for simultaneous MOF synthesis and co-immobilization of enzymes (GOx and HRP). The resultant enzymes-PADD@MOFs constructs showed a closely packed nanoarchitecture with enhanced substrate channeling. A transient time close to 0 s was observed, owing to a short diffusion path for substrates in a 2D spindle-shaped structure and their direct transfer from one enzyme to another. This enzyme cascade reaction system showed a 3.5-fold increase in catalytic activity in comparison to free enzymes. The findings provide a new insight into using polymer-directed MOF-based enzyme nanoarchitectures to improve catalytic efficiency and selectivity.

Enzyme Cascade Electrode Reactions with Nanomaterials and Their Applicability towards Biosensor and Biofuel Cells

“…In this study, they found that compared to subsequent immobilization, the one-step immobilization technique results in higher cascade enzymatic activity (Figure 7c). The utilization of PADD also increases the enzyme catalytic activity, 1.5 higher compared to the immobilized enzymes without PADD (Figure 7d) [84]. The covalent immobilization of enzymes on the surface of functionalized MOFs has also been described, in addition to immobilization through encapsulation or coprecipitation.…”

“…In this study, they found that compared to subsequent immobilization, the one-step immobilization technique results in higher cascade enzymatic activity (Figure 7c). The utilization of PADD also increases the enzyme catalytic activity, 1.5 higher compared to the immobilized enzymes without PADD (Figure 7d) [84].…”

Immobilized Dipeptidase in Manganese Ion-Loaded Polyethylenimine-Induced Calcium Phosphate Nanocrystals for Carnosine Synthesis

“…Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to confirm the purification of PepD. The PepD concentration was determined by the Bradford method …”

The role of protein folding in prenucleation clusters on the activity of enzyme@metal–organic frameworks

“…8,9 While little is known about a given enzyme environment and its exact conformation during MOF formation, enzymatic tertiary structure, location, and orientation within the crystal are known to play key roles in the activity of the immobilized enzyme. 10–12…”