Exploiting unique NP1 interface: Oriented immobilization via electrostatic and affinity interactions in a tailored...

Area of Science:

• Biochemistry
• Materials Science
• Enzyme Engineering

Background:

• Traditional enzyme immobilization methods often compromise enzyme activity and stability.
• Nuclease P1 (NP1) requires advanced immobilization for optimal performance.

Purpose of the Study:

• To develop a novel multi-level immobilization strategy for nuclease P1 (NP1).
• To fine-tune the surface microenvironment for enhanced enzyme performance.

Main Methods:

• Utilized molecular simulations to understand NP1 surface properties and dopamine adsorption.
• Employed a multi-level surface modification strategy involving dopamine and a positively charged environment.
• Conducted surface characterization and enzymatic testing to evaluate immobilization effectiveness.

Main Results:

• Achieved significantly improved enzyme activity (3590.0 U/mg), stability, and reusability (70% after 10 cycles).
• Demonstrated enhanced catalytic performance with an optimal Michaelis constant (K_m) of 34.0 mM and a maximum reaction rate of 5.5 mM min^-1.
• Confirmed optimization of the immobilized surface's physicochemical environment.

Conclusions:

• The developed strategy provides an efficient and stable platform for enzyme immobilization.
• This approach enhances enzyme catalytic efficiency and has potential in industrial biocatalysis, biomedical applications, and biosensors.