Presence and activity of endo-β-1,4-mannase, an important digestive carbohydrase within the digestive fluid of terrestrial crustaceans

Within the midgut gland of the Christmas Island red crab, Gecarcoidea natalis, a single transcript for a GH5_10 endo-β-1,4-mannase had the highest expression out of all of the carbohydrase enzymes (Gan et al. in Mar Biotechnol 20:654-665, 2018). The activity, and potential digestive importance of this hemicellulase, compared with other carbohydrases, has yet to be established. The digestive fluid of G. natalis contained substantial endo-β-1,4-mannase activities (630 ± 55 (6) nmol reducing sugars. min^-1. mg^-1 protein). It was present as a single isozyme of 66.3 ± 0.7 kDa (n = 6). Endo-β-1,4-mannase activities were higher than that for lichenase and endo-β-1,4-glucanase but lower than that for β-1,3-glucanase and amylase. The digestive fluid was able to hydrolyse, galactomannan, into its component monosaccharides. Hence, this confirms expression data that this enzyme is one of the most important digestive cellulases/ hemicellulases. Expression of GH5_10 endo-β-1,4-mannase mRNA was consistent with that of a digestive enzyme, as it was expressed in the digestive midgut gland but not in muscle and gill. Endo-β-1,4-mannase activities were also present within the digestive fluid of the terrestrial hermit crabs, Coenobita perlatus and Coenobita brevimanus. Endo-β-1,4-mannase activities (1351 ± 136 (n=3) nmol reducing sugars. min^-1 mg^-1 protein for C. perlatus. 665 ± 32 n=(5) nmol reducing sugars. min^-1 mg^-1 protein for C. brevimanus) were higher than that for endo-β-1,4-glucanase and amylase but were lower than β-1,3-glucanase activities. Animals within the terrestrial hermit crab family, Coenobitidae consume legume and palm seeds which contain substantial amounts of mannan. Hence, high endo-β-1,4-mannase activities suggest that digestion of mannan within these species may represent an important source of carbohydrate.