Construction and biochemical characterization of a novel hybrid alginate lyase with high activity by module recombination to prepare alginate oligosaccharides

Abstract

Alginate lyases are essential tools to prepare alginate oligosaccharides with various biological activities. However, alginate lyases with excellent properties such as high activity and good thermal stability are still in shortage. Therefore, it is crucial to exploit new alginate lyases with high activity and polysaccharide-degrading efficiency for alginate oligosaccharide preparation. Herein, we proposed to construct a novel hybrid alginate lyase with improved property by module recombination. The hybrid alginate lyase, designated as Aly7C, was successfully constructed by recombining the carbohydrate binding module (CBM) of Aly7A with the catalytic module of Aly7B. Interestingly, the hybrid enzyme Aly7C exhibited higher activity than the catalytic domain. Moreover, it could degrade sodium alginate, polyM and polyG into oligosaccharides with degrees of polymerization (Dps) 2–5, which exhibit perfect product specificity. This work provides a new insight into well-defined generation of alginate oligosaccharides with associated CBMs and enhances the understanding of functions of the modules.