Dual-Site Cooperative Modification Strategy Empowers Cellulase: Performance Optimization and pH-Sensitive Recovery

Area of Science:

• Biotechnology
• Biochemistry
• Enzyme Engineering

Background:

• Cellulose utilization via enzymatic hydrolysis is crucial for resource recycling and achieving carbon neutrality.
• Developing cellulase with enhanced activity and recyclability is a key research area.

Purpose of the Study:

• To construct a modified cellulase, Mal-Cell-FA, with pH-sensitive recovery properties.
• To improve the efficiency and recyclability of cellulase for cellulose hydrolysis.

Main Methods:

• Utilized transition state theory and dynamic amide bonds for enzyme modification.
• Employed aromatic aldehyde poly(ethylene glycol) for dual-site cooperative modification of the enzyme's catalytic and binding domains.

Main Results:

• The modified cellulase (Mal-Cell-FA) exhibited a 138% increase in activity compared to natural cellulase.
• Mal-Cell-FA demonstrated excellent recyclability, retaining over 80% of its activity after 5 cycles via pH regulation.
• The enhanced enzymatic activity is attributed to improved substrate accessibility.

Conclusions:

• The developed Mal-Cell-FA shows significant potential for efficient cellulose utilization.
• This strategy offers a novel approach to enhance enzyme performance and promote sustainable practices.