Enhancing the Catalytic Properties of a Novel Bifunctional κ-Carrageenanase through Spontaneous Self-Cyclization
Zhe Wang1,2,3, Suxue Chen1,2,3, Yan Li1,2,3
1State Key Laboratory of Marine Food Processing and Safety Control, College of Food Science and Engineering, Ocean University of China, Qingdao 266404, P. R. China.
Abstract:
Furcellaran is a distinct carrageenan polysaccharide. The unique oligosaccharides produced by its degradation exhibit lower sulfate group content and play a crucial role in elucidating the structure-activity relationship of carrageenan oligosaccharides. In this study, a novel κ-carrageenanase OUC-CgkA-Sn from Stieleria neptunia was identified to display dual substrate hydrolysis activity against κ-carrageenan and furcellaran. Significantly, the furcellaran-degrading activity of OUC-CgkA-Sn was 1.77-fold higher than that toward κ-carrageenan, and the primary degradation product was desulfurized κ-neocarrahexaose. To enhance its catalytic properties, the SpyTag/SpyCatcher system was employed to construct the fusion enzyme CKT, which is capable of spontaneous self-cyclization in vitro. After cyclization, CKT exhibited a dual enhancement in stability and enzyme activity simultaneously: 193.61% higher half-life and 195.21% greater enzyme activity. Spatial structure analysis revealed that this enhancement stemmed from increased structural stability and strengthened substrate interactions. As a result, CKT holds great promise for the efficient preparation of structurally diverse carrageenan oligosaccharides.
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