A novel CBM serving as a module for efficiently decomposing xanthan by modifying the processivity of hydrolase

The inefficient decomposition of polysaccharides, particularly branched polysaccharides limits their large-scale industrial applications. Further understanding and modification of glycoside hydrolases (GHs) processivity is expected to overcome this limitation. Here, a novel xanthan-binding CBM (MiXBM), which was supposed to alter the processivity of GHs, was systematically characterized. Phylogeny and structure analyses indicated that MiXBM is closely related to putative polysaccharide side chain-binding modules. Quantitative binding assays further revealed that MiXBM probably has a high affinity for xanthan side chain via a variable loop site. Moreover, catalytic performance demonstrated that xanthanase chimeras containing MiXBM promote highly efficient hydrolysis of xanthan because of improved substrate accessibility. Notably, MiXBM was observed to enhance the processivity of xanthanase, owing to its high substrate affinity to the repeating unit xanthan. Furthermore, sequential hydrolysis of xanthan by xanthanases with varying processivity resulted in significantly increased hydrolytic efficiency and focused oligoxanthans array. These results expand understanding of CBM-substrate recognition and shed light on efficient degradation of other regularly branched polysaccharides using modified GHs.