Endogenous production of endo-β-1,4-glucanase by decapod crustaceans

The potential ability to produce cellulase enzymes endogenously was examined in decapods crustaceans including the herbivorous gecarcinid land crabs <i>Gecarcoidea natalis</i> and <i>Discoplax hirtipes</i>, the amphibious freshwater crab <i>Austrothelphusa transversa</i>, the terrestrial hermit crab, <i>Coenobita variabilis</i> the parastacid crayfish <i>Euastacus</i>, and the crayfish <i>Cherax destructor</i>. The midgut gland of both <i>G. natalis</i> and <i>D. hirtipes</i> contained substantial total cellulase activities and activities of the cellulase enzymes endo-<i>β</i>-1,4-glucanase and <i>β</i>-glucosidase. With the exception of total cellulase and <i>β</i>-glucosidase from <i>D. hirtipes</i>, the enzyme activities within the midgut gland were higher than those within the digestive juice. Hence, the enzyme activities appear to reside predominantly within midgut gland, providing indirect evidence for endogenous synthesis of cellulase enzymes by this tissue. A 900 bp cDNA fragment encoding a portion of the endo-<i>β</i>-1,4-glucanase amino acid sequence was amplified by RT-PCR using RNA isolated from the midgut gland of <i>C. destructor</i>, <i>Euastacus</i>, <i>A</i>. <i>transversa</i> and <i>C. variabilis</i>. This provided direct evidence for the endogenous production of endo-<i>β</i>-1,4-glucanase. The 900 bp fragment was also amplified from genomic DNA isolated from the skeletal muscle of <i>G</i>. <i>natalis</i> and <i>D. hirtipes</i>, clearly indicating that the gene encoding endo-<i>β</i>-1,4-glucanase is also present in these two species. As this group of evolutionary diverse crustacean species possesses and expresses the endo-<i>β</i>-1,4-glucanase gene it is likely that decapod crustaceans generally produce cellulases endogenously and are able to digest cellulose.<br />