A novel chondroitin AC lyase from Pedobacter xixiisoli: Cloning, expression, characterization and the application in the preparation of oligosaccharides

Chondroitin AC lyase can efficiently hydrolyze chondroitin sulfate (CS) to low molecule weight chondroitin sulfate, which has been widely used in clinical therapy, including anti-tumor, anti-oxidation, hypolipidemic, and anti-inflammatory. In this work, a novel chondroitin AC lyase from Pedobacter xixiisoli (PxchonAC) was cloned and overexpressed in Escherichia coli BL21 (DE3). The characterization of PxchonAC showed that it has specific activities on chondroitin sulfate A, Chondroitin sulfate C and hyaluronic acid with 428.77, 270.57, and 136.06 U mg(-1), respectively. The K-m and V-max of PxchonAC were 0.61 mg mL(-1) and 670.18 U mg(-1) using chondroitin sulfate A as the substrate. The enzyme had a half-life of roughly 660 min at 37 degrees C in the presence of Ca2+ and remained a residual activity of 54 % after incubated at 4 degrees C for 25 days. Molecular docking revealed that Asn123, His223, Tyr232, Arg286, Arg290, Asn372, and Glu374 were mainly involved in the substrate binding. The enzymatic hydrolysis product was analyzed by gel permeation chromatography, demonstrating PxchonAC could hydrolyze CS efficiently.

Automated summary

Chondroitin AC lyase from Pedobacter xixiisoli efficiently hydrolyzes chondroitin sulfate to low molecular weight chondroitin sulfate, with specific activities on various substrates and good stability under certain conditions. Molecular docking analysis revealed key residues involved in substrate binding, and gel permeation chromatography confirmed the efficient hydrolysis capability of PxchonAC.