Expression, Purification and Characterization of Chondroitinase AC II from Marine Bacterium <i>Arthrobacter</i> sp. CS01
Yangtao Fang,
Suxiao Yang,
Xiaodan Fu,
Wancui Xie,
Li Li,
Zhemin Liu,
Haijin Mou,
Changliang Zhu
Affiliations
Yangtao Fang
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Suxiao Yang
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Xiaodan Fu
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Wancui Xie
Shandong Provincial Key Laboratory of Biochemical Engineering, College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao 266042, China
Li Li
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Zhemin Liu
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Haijin Mou
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Changliang Zhu
Laboratory of Applied Microbiology, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
Chondroitinase (ChSase), a type of glycosaminoglycan (GAG) lyase, can degrade chondroitin sulfate (CS) to unsaturate oligosaccharides, with various functional activities. In this study, ChSase AC II from a newly isolated marine bacterium Arthrobacter sp. CS01 was cloned, expressed in Pichia pastoris X33, purified, and characterized. ChSase AC II, with a molecular weight of approximately 100 kDa and a specific activity of 18.7 U/mg, showed the highest activity at 37 °C and pH 6.5 and maintained stability at a broad range of pH (5–7.5) and temperature (below 35 °C). The enzyme activity was increased in the presence of Mn2+ and was strongly inhibited by Hg2+. Moreover, the kinetic parameters of ChSase AC II against CS-A, CS-C, and HA were determined. TLC and ESI-MS analysis of the degradation products indicated that ChSase AC II displayed an exolytic action mode and completely hydrolyzed three substrates into oligosaccharides with low degrees of polymerization (DPs). All these features make ChSase AC II a promising candidate for the full use of GAG to produce oligosaccharides.
