Functional Analysis of a Novel ß-(1,3)-Glucanase from Corallococcus sp. Strain EGB Containing a Fascin-Like Module.

ABSTRACT

A novel ß-(1,3)-glucanase gene designated lamC, cloned from Corallococcus sp. strain EGB, contains a fascin-like module and a glycoside hydrolase family 16 (GH16) catalytic module. LamC displays broad hydrolytic activity toward various polysaccharides. Analysis of the hydrolytic products revealed that LamC is an exo-acting enzyme on ß-(1,3)(1,3)- and ß-(1,6)-linked glucan substrates and an endo-acting enzyme on ß-(1,4)-linked glucan and xylan substrates. Site-directed mutagenesis of conserved catalytic Glu residues (E304A and E309A) demonstrated that these activities were derived from the same active site. Excision of the fascin-like module resulted in decreased activity toward ß-(1,3)(1,3)-linked glucans. The carbohydrate-binding assay showed that the fascin-like module was a novel ß-(1,3)-linked glucan-binding module. The functional characterization of the fascin-like module and catalytic module will help us better understand these enzymes and modules.IMPORTANCE In this report of a bacterial ß-(1,3)(1,3)-glucanase containing a fascin-like module, we reveal the ß-(1,3)(1,3)-glucan-binding function of the fascin-like module present in the N terminus of LamC. LamC displays exo-ß-(1,3)/(1,6)-glucanase and endo-ß-(1,4)-glucanase/xylanase activities with a single catalytic domain. Thus, LamC was identified as a novel member of the GH16 family.