Enzymatic properties of a multi-specific β-(1,3)-glucanase from Corallococcus sp. EGB and its potential antifungal applications

The lamC gene encoding a novel β-(1,3)-glucanase was cloned from Corallococcus sp. EGB and successfully expressed in the industrial yeast Pichia pastoris. The mature protein without the initial 26 residues of signal peptide, designated LamC₂₇, was found to be composed of fascin-like module and laminarinase-like catalytic module. The purified recombinant enzyme (rLamC₂₇) with a calculated molecular mass of 45.3 kDa displays activities toward a broad range of β-linked polysaccharides, including laminarin, curdlan, pachyman, lichenan, and CMC. Enzymological characterization showed that rLamC₂₇ performes its optimal activity under the condition of 45 °C and pH 7.0, respectively, and preferentially catalyzes the hydrolysis of glucans with a β-1,3-linkage, which is similar to the LamC previously expressed in E. coli. TherLamC₂₇ enzyme was activated by Mn²⁺ and Ba²⁺, while it was inhibited by Cu²⁺, Zn²⁺, and Co²⁺. Moreover, rLamC₂₇ was strongly inhibited by 10 mM EDTA with 7.5% of its original activity remiaining, and weakly by SDS and Triton X-100. In antifungal assay, rLamC₂₇ was conformed to possess lytic and antifungal activity against rice blast fungus. Specifically, a significant decrease germ tube and appressorium formation ratios from 94% to 59% and 97%–51%, respectively, were observed following exposure to rLamC₂₇. H₂DCFDA and CFW staining further demonstrated that the fungistasis capability of rLamC₂₇ could be contributed by its ability to hydrolyze components of the cell wall. All these favorable properties indicate a promising potential for using rLamC₂₇ as a biological antifungal agent in areas such as plant protection and food preservation.