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Modification and application of highly active alkaline pectin lyase
Pi-Wu Li1,2, Jun Ma2, Xiao-Feng Wei2
1State Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology, Jinan, 250353, Shandong, Republic of China.
AMB Express
|October 9, 2022
Summary
Researchers enhanced alkaline pectate lyase (BacPelA) from Bacillus clausii for industrial applications. The modified enzyme, PGLA-rep4, shows improved catalytic activity, stability at high pH and temperature, and suitability for textile and paper processing.
Area of Science:
- Biotechnology
- Enzymology
- Industrial Microbiology
Background:
- Alkaline pectate lyase shows industrial potential in textiles, pulp, paper, and food sectors.
- Bacillus clausii pectin lyase (BacPelA) was identified as a highly expressed enzyme candidate.
Purpose of the Study:
- To enhance the catalytic activity and industrial applicability of BacPelA.
- To characterize the modified pectin lyase for optimal performance under industrial conditions.
Main Methods:
- Enzyme engineering via fragment replacement to modify BacPelA.
- Expression of the modified enzyme in Escherichia coli BL21(DE3).
- Biochemical characterization including pH, temperature optima, substrate specificity, and stability studies.
- Molecular dynamics simulations to analyze structural stability.
Main Results:
- The modified pectin lyase, PGLA-rep4, exhibited significantly improved catalytic activity.
- Optimal activity was observed at pH 11.0 and 70 °C, with superior cleavage of methylated pectin.
- Purified PGLA-rep4 showed high specific enzyme activity (822.9 U/mg) and stability in the presence of metal ions.
- Molecular dynamics simulations indicated structural stability, with increased fluctuations at the LYS168 active site compared to wildtype.
Conclusions:
- Fragment replacement successfully enhanced the catalytic activity and stability of alkaline pectate lyase.
- PGLA-rep4 demonstrates robust enzymatic properties suitable for high-temperature and high-pH industrial processes.
- The engineered enzyme is a promising candidate for applications such as textile degumming and pulp/paper processing.
