High-Throughput Screening Identifies Anionic Polymer Supports that Improve Enzyme Activity at Low pH and High...

Area of Science:

• Biochemistry
• Polymer Science
• Enzyme Engineering

Background:

• Enzyme performance is often limited by suboptimal pH and high temperatures.
• Enzyme immobilization on polymer supports can improve stability and activity by controlling the local microenvironment.
• Multicomponent polymer brushes offer tunable properties for enzyme stabilization.

Purpose of the Study:

• To investigate the stabilization of lipase A using multicomponent random copolymer brushes.
• To evaluate the effect of different monomer properties (anionic, cationic, neutral, aromatic) on enzyme stability.
• To understand the mechanisms behind enhanced enzyme performance under harsh conditions.

Main Methods:

• Combinatorial synthesis of random copolymer brushes with varied monomer compositions.
• High-throughput screening of immobilized lipase A activity and stability.
• In situ characterization of enzyme function at low pH and elevated temperatures.

Main Results:

• Anionic and neutral zwitterionic polymer supports significantly enhanced lipase A stability and activity.
• Temperature optima increased by up to 40 °C (to 80 °C), and maximum activity increased by over 300%.
• Cationic supports offered modest improvements, while anionic and zwitterionic supports showed substantial benefits.

Conclusions:

• Enzyme stabilization by charged polymers can occur through mechanisms beyond local pH control, such as preserving enzyme structure via electrostatic interactions.
• Negatively charged polymer moieties effectively stabilized the positively charged surface of lipase A.
• These findings highlight the potential of tailored polymer brushes for robust enzyme immobilization and applications.