Abstract
Alginate is a commercially important polysaccharide widely applied in various industries. Carbohydrate-binding proteins could be utilized as desirable tools in the investigation and further applications of polysaccharides. Few alginate-binding proteins have hitherto been characterized and reported. In the present study, a novel alginate-binding protein ABP_Wf, consisting of two "orphan" carbohydrate-binding modules, was cloned from a predicted alginate utilization locus of marine bacterium Wenyingzhuangia funcanilytica, and expressed in Escherichia coli. ABP_Wf exhibited a specific binding capacity to alginate, and the association constant (Ka) and affinity (KD) were 1.94 × 103 M-1s-1 and 1.16 × 10-6 M. It was confirmed that the binding capacity of ABP_Wf to alginate is attributed to its constituent CBM16 domain rather than the CBM44 domain. The potentials of ABP_Wf in the semi-quantitative detection and the in situ visualization of alginate were evaluated, which implied that ABP_Wf could be served as a promising tool for investigating alginate.