A Novel Chondroitin AC Lyase With Broad Substrate Specificity From Pedobacter rhizosphaerae : Cloning, Expression, and Characterization

• Published in: Frontiers in bioengineering and biotechnology Vol. 9; p. 808872
• Main Authors: Zhou, Li-Jian, Guo, Li-Bin, Wei, Wei, Lv, Zhi-Xiang, Zhang, Ye-Wang
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 23.12.2021
• Subjects: Bioengineering and Biotechnology; characterization; chondroitin AC lyase; cloning; expression; molecular docking; expression; molecular docking; chondroitin AC lyase; characterization; cloning
• ISSN: 2296-4185; 2296-4185
• DOI: 10.3389/fbioe.2021.808872

Chondroitin AC lyase (ChSaseAC) is one of the essential polysaccharides lyases in low molecular chondroitin sulfate production. In this work, a novel PrChSaseAC from was successfully cloned, expressed in . After optimizing the induction, the recombinant PrChSaseAC could be expressed efficiently at 0.1 mM IPTG, 25°C, and 12 h induction. Then, it was purified with Ni-NTA affinity chromatography. The characterization of the purified PrChSaseAC showed that it had high specific activity and good storage stability, which would favor the production of low molecular weight chondroitin sulfate. It also displayed activity toward chondroitin sulfate C and hyaluronic acid. PrChSaseAC had the highest activity at pH 7.5, 37°C, 10 mM Ca , and 5 mg/ml of chondroitin sulfate A. Molecular docking of substrate and enzyme showed the interactions between the enzyme and substrate; it revealed that the enzyme showed high activity to CS-A and hyaluronic acid, but lower activity to CS-C attributed to the structure of the binding pocket. The high stability and specific activity of the enzyme will benefit the industrial production or clinical treatment.