Related Experiment Video
Updated: Aug 13, 2026
05:26
Fabrication of Size-Controlled and Emulsion-Free Chitosan-Genipin Microgels for Tissue Engineering Applications
Published on: April 13, 2022
Rational Engineering of Chitosanase for Producing Higher-Degree-of-Polymerization Chitooligosaccharides and
Zhangyi Chen1, Jiawei Luo1, Chenglin Wu1,2
1Taizhou Key Laboratory of Biomass Functional Materials Development and Application, School of Life Science, Taizhou University, Taizhou318000, China.
Journal of Agricultural and Food Chemistry
|August 12, 2026
Summary
Engineered chitosanase Csn168 produces high-degree of polymerization chitooligosaccharides (COS). These functional COS show potent antibacterial activity against Staphylococcus aureus, offering a new strategy for functional biomaterial development.
Area of Science:
- Biochemistry
- Biotechnology
- Microbiology
Background:
- High-degree of polymerization chitooligosaccharides (COS) exhibit superior biological activities.
- Chitosanase Csn168 from Bacillus subtilis 168 typically produces low-DP COS.
Purpose of the Study:
- To engineer chitosanase Csn168 for increased production of high-DP COS.
- To evaluate the antibacterial properties of the resulting high-DP COS.
Main Methods:
- Rational design introducing three negatively charged mutations (L117E/G21D/T50E) into chitosanase Csn168.
- Analysis of COS product distribution using HPLC.
- Assessment of antibacterial activity against Staphylococcus aureus.
- Molecular dynamics simulations to investigate membrane interactions.
Main Results:
- The engineered mutant shifted product distribution from low-DP (GlcN)2 and (GlcN)3 to high-DP (GlcN)4 and (GlcN)5 (85.3%).
- High-DP COS achieved complete growth inhibition of Staphylococcus aureus at 8 g/L.
- Molecular dynamics simulations suggested enhanced membrane interactions for (GlcN)4/(GlcN)5.
Conclusions:
- A rational design strategy successfully modulated COS polymerization degree.
- Engineered chitosanase Csn168 produces functional high-DP COS with potent antibacterial activity.
- This approach advances the production of functional COS for potential applications.
