Functional Characterization of a Highly Efficient Endoglucanase from Bacillus licheniformis BJ2022 and Its Application in the Preparation of Low-Molecular-Weight Konjac Glucomannan

Endoglucanases may not only act on beta-1,4-linkages in beta-glucan but also target d-glucose in glucomannan, which can expand their utility in the preparation of prebiotics. In this study, a highly efficient endoglucanase (BlGH5) from Bacillus licheniformis BJ2022 was expressed and characterized. BlGH5 exhibited the highest activity at pH 7.0 and 60 degrees C. It maintained over 80% activity at pH 4.0-7.0 and 30-60 degrees C. BlGH5 specifically cleaved beta-1,4-glycosidic bonds linked to d-glucose. Site-directed mutagenesis results suggested that Arg91, Asn167, Glu168, Trp206, His228, Tyr230, Ser263, and Trp290 are key residues for its binding and catalytic activity. Moreover, BlGH5 displayed high activity against konjac glucomannan (KGM), indicating that BlGH5 could be used to prepare low-molecular-weight konjac glucomannan (KGM-L). Based on the physicochemical properties of KGM and KGM-L, KGM-L was characterized by reduced molecular weight and viscosity. Fecal fermentation experiments demonstrated that KGM and KGM-L could promote the production of short-chain fatty acids (SCFAs). Still, KGM-L was more conducive to the growth of the gut probiotics. In conclusion, we identified an endoglucanase for the degradation of KGM. Results indicated that KGM-L would have superior prebiotic effects. Thus, our study provides a basis for the future development and application of KGM-L as a prebiotic.